CN101862351A - Application of active parts of nutgall in preparing medicine for resisting ulcerative colitis - Google Patents

Abstract

The invention relates to an application of active parts of gallnut in preparing a medicament for resisting ulcerative colitis, wherein the active parts of the gallnut are extracted by an organic solvent after water extraction to obtain the active parts of the gallnut, which mainly comprise tannins; and performing gel chromatography extraction and separation on the effective part to obtain monomer compounds, such as gallic acid, m-digallic acid, gallic acid methyl ester, 1, 2, 3, 6-tetra-O-galloyl-beta-D-glucose, and 1, 2, 3, 4, 6-penta-O-galloyl-beta-D-glucose. Clinical results show that the active parts of the gallnut have obvious treatment effect on the ulcerative colitis, the total effective rate is 91.7%, the recent cure rate is 66.7%, the effective rate is 25%, and the ineffective rate is 8.3%. Pharmacological research shows that the active parts of the gallnut have obvious therapeutic action on the ulcerative colitis of the rat; can significantly increase SOD content in ulcer tissue, reduce MPO content, has obvious anti-inflammatory and analgesic effects, and has antibacterial effect on common pathogenic bacteria related to ulcerative colitis.

Description

Use of the effective part of gallaceae in the preparation of anti-ulcerative colitis medicine

technical field

The invention relates to the medicinal application of the effective parts of gallicia, in particular to the preparation of medicinals for treating ulcerative colitis.

Background technique

Gallalls are the larvae of Cynips gallae-tinctorice Oliv, an insect of the family Apiaceae, which parasitize on the twigs of Quercus infectoria Oliv. Gallatin is dry and cold in nature, bitter and astringent in taste, and has the functions of strengthening astringency, astringent, dampness, hemostasis, and anti-inflammatory. It is a commonly used medicinal material in Uighur medicine. The theory of Uyghur medicine believes that it can directly act on the intestinal wall to reduce the absorption and pinching force of the intestinal wall caused by colitis, astringe the intestines, reduce inflammation and diarrhea, open up intestinal resistance, and promote ulcer healing. Clinically used in the treatment of ulcerative colitis, the curative effect is remarkable. However, there is no report about the use of the effective part of the gallic acid as a medicine for preparing anti-ulcerative colitis.

Ulcerative colitis (UC), also known as nonspecific ulcerative colitis, is a chronic inflammatory disease of the rectum and colon of unknown cause, with abdominal pain, diarrhea, mucus bloody stool, and tenesmus as the main clinical manifestations. The lesions are mainly located in the mucosal layer of the colon, mainly ulcers, mostly involving the rectum and distal colon, and can also spread throughout the colon. Epidemiological data suggest that the incidence of ulcerative colitis has been increasing year by year both at home and abroad. In the past 10 years, the prevalence of ulcerative colitis in my country has increased by about 10 times compared with the 1950s. 20 European centers The survey shows that the annual incidence rate of ulcerative colitis is 11.2/100000. The disease can occur at any age and is related to the onset of colon cancer. However, it is difficult to cure, and it is listed as one of the modern intractable diseases by the World Health Organization.

Contents of the invention

The purpose of the present invention is to provide an effective part of gallaceae as the preparation of a medicine for treating ulcerative colitis. The effective part of gallaceae can be obtained by extracting with water and then an organic solvent, mainly including tannins; Then the effective parts are extracted and separated by gel chromatography to obtain monomeric compounds, including gallic acid, m-digallic acid, methyl gallate, 1,2,3,6-tetra-O-galloyl-β-D -glucose, 1,2,3,4,6-penta-O-galloyl-β-D-glucose. The clinical results show that the effective parts of Gallicia have obvious therapeutic effects on ulcerative colitis, the total effective rate is 91.7%, the short-term cure rate is 66.7%, the effective rate is 25%, and the ineffective rate is 8.3%. Pharmacological studies have shown that the effective part of gallaceae has obvious therapeutic effects on rat ulcerative colitis; it can significantly increase the content of SOD in ulcer tissue, reduce the content of MPO, and has obvious anti-inflammatory and analgesic effects, which are related to ulcerative colitis. The common pathogenic bacteria have antibacterial effect, and the effective part of the gall is a good medicine for the treatment of ulcerative colitis.

The use of the effective part of Gallia according to the present invention in the preparation of a medicine for treating ulcerative colitis, the extraction method of the effective part of Gallia is carried out according to the following steps:

a. Take the dried gallic medicinal material and crush it, pass through a No. 2 sieve, weigh 100g of medicinal material powder, add water to extract 3 times, add 5-15 times the amount of water each time, and extract for 20-120min;

b. Concentrate the extract under reduced pressure to 500-1500ml, extract the concentrate with ether until the ether layer is clarified, and discard the ether layer;

c. Extract the part of the water layer with ethyl acetate, recover the ethyl acetate, and dry it in vacuum at a temperature of 45-90°C to obtain 30-60% of the effective parts of the galls;

d. Separating the chemical components of the active part of gallic acid obtained in step c by using a conventional gel chromatography column to obtain monomeric compounds, including gallic acid, meta-digallic acid, methyl gallate, 1,2, 3,6-tetra-O-galloyl-β-D-glucose, 1,2,3,4,6-penta-O-galloyl-β-D-glucose.

The effective parts of Gallus adopt the 2000 edition pharmacopoeia appendix X B tannin content determination method, and the content of measuring tannin is greater than 50%.

On the basis of the clinical efficacy of gallaceae in treating ulcerative colitis, the present invention conducts a systematic study on gallacea: (1) focusing on the safety of gallacea extracts, general toxicology research is carried out on gallacea extracts. Rats were subjected to rectal administration acute toxicity test, rectal administration mucosal irritation test and long-term toxicity test. The results of acute toxicity test of rectal administration in rats show that the MTD of single rectal administration is more than 13g/kg, which is 245 times that of adult clinical dosage. There is no toxic reaction caused by the drug at this dose, and no pathological anatomy is found. Obvious pathological changes in rat organs. The results of rectal administration mucosal irritation test and long-term toxicity test in rats showed that the rectal mucosa hyperemia and redness caused by drugs were not seen in rats after rectal administration for many times, and the rectal mucosa of rats was not caused by drugs in histopathology. pathological changes. Gallium extract is non-irritating. Compared with the control group, there was no significant difference in each index between rats administered with 3 different doses of gallaceae extract, and no pathological changes related to drug effects were seen. Under these three different dosage conditions, the gallate extract has no obvious toxicity to rats after administration for 120 days. (2) In order to study the therapeutic effect of gall on the rat ulcerative colitis (UC) animal model, the rat UC model was prepared by the compound method of trinitrobenzenesulfonic acid and ethanol. Extract treatment, evaluation of rat colonic mucosal injury index (CMDI), detection of changes in body weight, intestinal weight index and immune organ weight index, detection of superoxide dismutase (SOD), myeloperoxidase ( MPO) activity. The test results showed that the ulcers of the animals in the high-dose gallate extract group were completely healed. Compared with the model group animals, the SOD in the high-dose and low-dose groups was significantly increased (P<0.01), and the MPO was significantly decreased (P<0.01). Gallic seed extract has obvious therapeutic effect on ulcerative colitis in rats. (3) Taking the total tannins of galls as an investigation index, screening tests were carried out on the extraction process. As a result, the galls were extracted by water and extracted with ether, and the water layer was extracted with ethyl acetate. The ethyl acetate was recovered, vacuum-dried, and the dry extract was obtained. The rate is higher than 40%, and the total tannin content is greater than 60%. It shows that this method can better enrich the total gallic tannins.

The effective part of the extracted gall is made into an oral preparation according to a conventional method, and the preparation is used as a preparation for treating ulcerative colitis.

The new anti-ulcerative colitis drug from the effective parts of gallaceae intends to use the abundant resources of gallaceae medicinal materials in nature to develop anti-ulcerative colitis Uyghur pharmaceutical preparations and new drugs. It is a preparation made from the effective part extracted from the gallic medicinal material - gallic total tannin. It is clinically used for the treatment of ulcerative colitis, and the clinical effect is remarkable. The preparation will be popularized and used as an effective drug for treating ulcerative colitis, and will bring good news to patients with ulcerative colitis to relieve pain.

Detailed ways

Example 1

a. Take the dried gallic medicinal material and crush it, pass it through a No. 2 sieve, weigh 100g of the medicinal material powder, add water to extract 3 times, add 5 times the amount of water each time, and extract for 20 minutes;

b. Concentrate the extract under reduced pressure to 500ml, extract the concentrated solution with ether until the ether layer is clarified, and discard the ether layer;

c. Extract the part of the water layer with ethyl acetate, recover the ethyl acetate, and dry it in vacuum at a temperature of 45° C. to obtain 30% effective parts of galls;

d. Use the gel chromatography column (model: Sephadex LH-20) to separate the chemical components of the effective part of gallia obtained in step c, and carry out gradient elution with water and 20%-100% methanol to obtain 40%-70 % methanol eluent extract Ⅰ and 70%-100% methanol eluent extract Ⅱ, and then the extract Ⅰ and Ⅱ were separated by type: MCI-GEL CHP-20P gel chromatography column, and water and 20%-80% methanol gradient elution; extract Ⅰ obtained compound gallic acid, m-digallic acid and methyl gallate; extract Ⅱ obtained extract Ⅲ and Extract Ⅳ, then separate extract Ⅲ and extract Ⅳ with model: Sephadex LH-20 gel chromatography column, and use water and 20%-100% methanol for gradient elution, and 60% of extract Ⅲ Compound 1,2,3,6-tetra-O-galloyl-β-D-glucose was obtained in 100% methanol eluent; it was obtained in 40%～60% methanol eluate of extract IV Compounds 1, 2, 3, 4, 6-penta-O-galloyl-β-D-glucose were obtained.

The effective parts of the galls adopt the 2000 edition pharmacopoeia appendix Ⅹ B tannin content determination method, and the measured tannin content is greater than 50%.

Example 2

a. Take the dried gallic medicinal material and pulverize it, pass through a No. 2 sieve, weigh 100g of the medicinal material powder, add water to extract 3 times, add 10 times the amount of water each time, and extract for 50 minutes;

b. Concentrate the extract under reduced pressure to 800ml, extract the concentrated solution with ether until the ether layer is clarified, and discard the ether layer;

c. Extract the part of the water layer with ethyl acetate, recover the ethyl acetate, and dry it in vacuum at a temperature of 65° C. to obtain 45% of the effective part of the galls;

d. Use the gel chromatography column (model: Sephadex LH-20) to separate the chemical components of the effective part of gallia obtained in step c, and carry out gradient elution with water and 20%-100% methanol to obtain 40%-70 % methanol eluent extract Ⅰ and 70%-100% methanol eluent extract Ⅱ, and then the extract Ⅰ and Ⅱ were separated by type: MCI-GEL CHP-20P gel chromatography column, and water and 20%-80% methanol gradient elution; extract Ⅰ obtained compound gallic acid, m-digallic acid and methyl gallate; extract Ⅱ obtained extract Ⅲ and Extract Ⅳ, then separate extract Ⅲ and extract Ⅳ with model: Sephadex LH-20 gel chromatography column, and use water and 20%-100% methanol for gradient elution, and 60% of extract Ⅲ Compound 1,2,3,6-tetra-O-galloyl-β-D-glucose was obtained in -100% methanol eluent; it was obtained in 40%-60% methanol eluate of extract IV Compounds 1, 2, 3, 4, 6-penta-O-galloyl-β-D-glucose were obtained.

The effective parts of the galls adopt the 2000 edition pharmacopoeia appendix Ⅹ B tannin content determination method, and the measured tannin content is greater than 50%.

Example 3

a. Take the dried gallic medicinal material and crush it, pass it through a No. 2 sieve, weigh 100g of medicinal material powder, add water to extract 3 times, add 15 times the amount of water each time, and extract for 120min;

b. Concentrate the extract under reduced pressure to 1500ml, extract the concentrated solution with ether until the ether layer is clarified, and discard the ether layer;

c. Extract the water layer with ethyl acetate, recover the ethyl acetate, and vacuum dry at a temperature of 90°C to obtain 60% of the effective parts of the galls;

d. Use the gel chromatography column (model: Sephadex LH-20) to separate the chemical components of the effective part of gallia obtained in step c, and carry out gradient elution with water and 20%-100% methanol to obtain 40%-70 % methanol eluent extract Ⅰ and 70%-100% methanol eluent extract Ⅱ, and then the extract Ⅰ and Ⅱ were separated by type: MCI-GEL CHP-20P gel chromatography column, and water and 20%-80% methanol gradient elution; extract Ⅰ obtained compound gallic acid, m-digallic acid and methyl gallate; extract Ⅱ obtained extract Ⅲ and Extract Ⅳ, then separate extract Ⅲ and extract Ⅳ with model: Sephadex LH-20 gel chromatography column, and use water and 20%-100% methanol for gradient elution, and 60% of extract Ⅲ Compound 1,2,3,6-tetra-O-galloyl-β-D-glucose was obtained in 100% methanol eluent; it was obtained in 40%～60% methanol eluate of extract IV Compounds 1, 2, 3, 4, 6-penta-O-galloyl-β-D-glucose were obtained.

The effective parts of the galls adopt the 2000 edition pharmacopoeia appendix Ⅹ B tannin content determination method, and the measured tannin content is greater than 50%.

Example 4

a. Take the dried gallic medicinal material and pulverize it, pass it through a No. 2 sieve, weigh 100g of the medicinal material powder, add water to extract 3 times, add 12 times the amount of water each time, and extract for 80 minutes;

b. Concentrate the extract under reduced pressure to 1100ml, extract the concentrated solution with ether until the ether layer is clarified, and discard the ether layer;

c. Extract the part of the water layer with ethyl acetate, recover the ethyl acetate, and dry it in vacuum at a temperature of 80°C to obtain 50% of the effective part of the galls;

d. The active part of gallia obtained in step c is separated from the chemical components by conventional gel chromatography (model: Sephadex LH-20), and gradient elution is carried out with water and 20%-100% methanol to obtain 40%- The extract I of 70% methanol eluent and the extract II of 70%-100% methanol eluate, and then the extract I and II were separated by type: MCI-GEL CHP-20P gel chromatography column, and Gradient elution with water and 20%-80% methanol; extract I obtained compound gallic acid, m-digallic acid and methyl gallate; extract II obtained extract III of 30%-60% methanol eluent and extract IV, and extract extract III and extract IV were separated with a model: Sephadex LH-20 gel chromatography column, and carried out gradient elution with water and 20%-100% methanol, at 60% of extract III Compound 1,2,3,6-tetra-O-galloyl-β-D-glucose was obtained in the methanol eluent of %-100%; in the 40%-60% methanol eluent of extract IV Compound 1,2,3,4,6-penta-O-galloyl-β-D-glucose was obtained.

The effective parts of the galls adopt the 2000 edition pharmacopoeia appendix Ⅹ B tannin content determination method, and the measured tannin content is greater than 50%.

The effective part of gallaceae is used as a medicine for the treatment of anti-ulcerative colitis. The effective part of gallacea can be obtained by extracting with water and then extracted with an organic solvent, mainly including tannins; the effective part can be extracted and separated by gel chromatography to obtain the monomer compounds, including gallic acid, m-digallic acid, methyl gallate, 1,2,3,6-tetra-O-galloyl-β-D-glucose, 1,2,3,4,6-penta- For O-galloyl-β-D-glucose, the effective part of the extracted gall is prepared into an oral preparation according to a conventional method.

Example 5 Experimental Research on the Effective Parts of Gallatin in the Preparation and Treatment of Anti-ulcerative Colitis

Abstract: Objective: To study the therapeutic effect of gallic seed extract on rat ulcerative colitis (UC) animal model. METHODS: Rat UC models were prepared by compound method of trinitrobenzenesulfonic acid and ethanol. After the successful model establishment was confirmed by pathological sections, they were randomly divided into high-dose and low-dose gallile extract groups, model group and sulfasalazine group. After 20 days of intragastric administration, evaluate the colonic mucosal injury index (CMDI); detect the changes in body weight, intestinal weight index and immune organ weight index; detect superoxide dismutase (SOD), myeloperoxide Enzyme (MPO) activity. Results: The ulcers of the animals in the high-dose gallacea extract group were completely healed. Compared with the model group, the SOD in the high-dose and low-dose groups was significantly increased (P<0.01), and the MPO was significantly decreased (P<0.01). CONCLUSION: The extract of Gallus chinensis has obvious therapeutic effect on ulcerative colitis in rats.

The present invention establishes a colitis model in rats, observes its therapeutic effect and detects relevant indexes after administration of the gallate extract, so as to understand its mechanism of action for treating ulcerative colitis.

Experimental materials and methods

Animals and Groups

50 SPF grade SD rats, half male and half female, 200-250 g, provided by: Medical Animal Experiment Center of Nanjing Military Region of the Chinese People's Liberation Army, license number: SCXK (Su) 2003-0004. They were randomly divided into blank control group, model group, positive control group and high and low dose groups of gallate extract, with 10 rats in each group.

Drugs and Reagents

The extract of the effective parts of Galla, provided by the Uygur Medical Research Institute of Xinjiang Uygur Autonomous Region, batch number: 090926; 2,4,6-trinitrobenzenesulfonic acid (TNBS) is a product of Sigma Company, 50% volume fraction, batch number: 046K5008; anhydrous Ethanol (AR) is a product of Nanjing Chemical Reagent Co., Ltd., batch number: 061110961; ether (AR) is a product of Nanjing Chemical Reagent Co., Ltd., batch number: 060610483; Sulfasalazine Enteric-coated Tablets is a product of Shanghai Sanwei Pharmaceutical Co., Ltd. : H31020450, batch number: 200701C03; SOD kit (for measuring total superoxide dismutase), batch number: 20070516; myeloperoxidase (MPO) test box, batch number: 20070516, all of the above test kits were provided by Nanjing Jiancheng Institute of Bioengineering product

model preparation

60 SD rats, half male and half male, weighing 200-250g in groups, were marked for weight, fasted for one day, and had free access to water. Modeling: Mix 50% (V/V) ethanol and 5% TNBS in equal amounts to prepare a modeling solution. Anesthetize the rat with ether and fix it. Insert a section of baby catheter with a diameter of 2.0mm and a length of about 10cm into the anus of the rat to a depth of about 8cm. Slowly inject 1ml/piece of a mixed solution of TNBS and ethanol. Rats were slightly tilted on their backs to prevent fluid leakage. In the following 2-3 days, the physiological changes of the rats were observed. Diarrhea, mucus, pus and blood were successfully modeled.

medication

From the 7th day after the preparation of the model, it was administered by intragastric administration, once a day, for a total of 20 days. The blank control group and the model group were given 10ml/kg normal saline by intragastric administration. Positive control group: sulfasalazine enteric-coated tablets 100 mg/kg, and the administration doses of the high-dose and low-dose gallile extract groups were 1.68 g/kg and 0.34 g/kg, respectively.

Preparation of tissue samples and index detection

On the 21st day after the administration of the rats, the rats were sacrificed, and the colon tissue was collected for morphological observation and colon tissue injury score (CMDI); the colon was clipped and weighed to calculate the intestinal mass index. Part of the colon was randomly selected, fixed with 10% (V/V) formalin solution, and then the lesion and three sections of intestine before and after were taken, washed with running water, routinely prepared for sectioning, and stained with HE. At the same time, the spleen and thymus were removed, weighed, and their visceral index calculated. Blood was collected from the eyeballs of the rats before they were sacrificed, the serum was separated, centrifuged at 3000r/min for 10 minutes, stored at -4°C for later use, and the activities of SOD and MPO were determined according to the kit instructions.

statistical processing

SPSS11.5 software was used to process statistical data, and t test and F analysis (q test) were used to analyze the mean values of the above-mentioned groups of experiments.

Experimental results

Effects of effective fractions of gallatum on colonic injury in rats with ulcerative colitis induced by TNBS

CMDI is the main index to reflect the degree of colon injury, and there is a significant difference between the high-dose group and positive drug group and the model group (P<0.01), and there is no statistical difference between the low-dose group and the model group.

Table 1 Effects of effective fractions of gallaceae on colonic injury in rats with ulcerative colitis induced by TNBS

Compared with the model group: ^▲ P＜0.05 ^▲▲ P＜0.01.

Effects of effective fractions of gallaceae on the intestinal mass index of rats with ulcerative colitis induced by TNBS:

Compared with the normal group, there was no significant difference in the intestinal weight index of the high-dose effective parts of gallaceae extract group and the positive drug group (P>0.05), which was significantly lower than that of the model group (P<0.01); There was a significant difference between the two groups (P<0.01), which was significantly lower than that of the model group (P<0.05).

Table 2 Effects of effective fractions of gallaceae on the intestinal mass index of rats with ulcerative colitis induced by TNBS

Compared with the normal group: **P<0.05, **P<0.01; compared with the model group: ^▲ P<0.05 ^▲▲ P<0.01

Determination of Relative Weight of Rat Immune Organs

Compared with the control group, there was no significant difference in the immune organs of the positive drug group, the high-dose and low-dose gallicula extract groups (P>0.05); there was no statistical significance between each group and the model group (P>0.05).

Table 3 Effects of effective fractions of gallaceae on the immune organs of rats with ulcerative colitis induced by TNBS

Compared with the normal group: *P<0.05**P<0.01; compared with the model group: ^▲ P<0.05 ^▲▲ P<0.01.

Effects of effective fractions of gallia on serum MPO activity in rats with UC induced by TNBS:

Compared with the model group, the MPO activity of the high-dose group and the positive drug group decreased significantly (P<0.01), and there was no statistical significance between the low-dose group and the model group (P>0.05); SOD activity was significantly increased in high and low dose groups (P<0.01).

Table 4 Effects of effective fractions of gallaceae on the activity of MPO and SOD in serum of rats with ulcerative colitis induced by TNBS

Compared with the normal group: *P<0.05**P<0.01; compared with the model group: ^▲ P<0.05 ^▲▲ P<0.01.

Pathological examination results

Blank control group: the colon tissue of each mouse is lined with a single-layer columnar epithelium, the mucosa is complete, the goblet cells are abundant, the mucosa is scattered with lymphoplasmacytic infiltration, lymphoid follicles can be seen under the mucosa in some areas, and the muscular layer and serosa layer have no obvious abnormalities . Model group: the epithelium of the colonic mucosa of 9 mice basically degenerated and fell off, replaced by large areas of necrosis and inflammatory exudate, the submucosal was highly edematous, and a large number of inflammatory cell infiltration was seen in the submucosal, muscular layer, serosal layer and outside the serosa , mainly neutrophils. In the other mouse, except for a slight increase in inflammatory cells in the colonic mucosa, the morphology of the colonic tissue of the other mouse was roughly similar to that of the blank control group. Positive drug administration group: the colon histology of each mouse was basically the same as that of the blank control group, lined with a single layer of columnar epithelium, the mucosa was complete, the goblet cells were abundant, scattered lymphoplasmacytic infiltration in the mucosa, and lymphoid follicles could be seen under the mucosa in some areas. There were no obvious abnormalities in the muscular layer and serosal layer. Low-dose administration group: among them, 7 cases of rat colonic mucosa saw scattered punctate epithelial degeneration and shedding, submucosal and muscular layer saw more acute and chronic inflammatory cell infiltration, and lymphatic follicles formed filling ulcers, and the remaining 3 cases of rat colonic mucosa The morphology was basically the same as that of the blank control group. The mucosa was intact, with abundant goblet cells, scattered lymphoplasmacytic infiltration in the mucosa, and lymphoid follicles under the mucosa in some areas. There were no obvious abnormalities in the muscular layer and serosa layer. In the high-dose administration group: in the colonic mucosa of 8 cases of mice, epithelial degeneration and exfoliation in multiple large areas of the mucosa were seen, and there were many inflammatory exudates and necrosis. Predominantly neutrophils. The morphology of the colonic mucosa of the remaining 2 mice was basically the same as that of the blank control group. The mucosa was complete, with abundant goblet cells, scattered lymphoplasmacytic infiltration in the mucosa, and lymphoid follicles under the mucosa in some areas. There were no obvious abnormalities in the muscular layer and serosa layer.

discuss

At present, the commonly used drugs for the treatment of ulcerative colitis mainly include hormones (such as corticosteroids), aminosalicylic acids (such as: SASP, 5-ASA), immunosuppressants (such as methotrexate), calcium ion antagonists, heparin, etc. , but the effect is not good, and there are many adverse reactions, and it is easy to relapse after stopping the drug. Traditional Chinese medicine treatment mostly treats the root by regulating the spleen and stomach, nourishing the liver and nourishing the kidney, clearing away heat and detoxification, promoting blood circulation and removing stasis, and strengthening the intestines to treat the symptoms. In terms of treatment methods, oral administration of traditional Chinese medicine, combined use of traditional Chinese and Western medicine, and retention enema of traditional Chinese medicine have all received certain curative effects in clinical practice. It is particularly worth mentioning that the results of oral administration of traditional Chinese medicine and oral administration of western medicine show that the curative effects of traditional Chinese medicine and traditional Chinese medicine are better than those of simple oral administration. The use of western medicine shows that traditional Chinese medicine has certain advantages in treating the disease. At the same time, the retention enema of traditional Chinese medicine has the same curative effect as internal medicine, and the operation is simple. Gallic seeds are dry and cold in nature, bitter in taste, and have the functions of firming astringent, astringent, dampness, hemostasis, and anti-inflammation. Studies have shown that gallic seeds contain tannin, flavonoids, gallic acid and other ingredients, and the tannin content of the tannin extract exceeds 50%. . Pharmacological studies have proved that the extract of gall seeds has a strong inhibitory and killing effect on Staphylococcus aureus, Escherichia coli, β-hemolytic streptococcus, etc.; it can also significantly inhibit acute and chronic inflammation, improve the body's non-specific immune function, It has the effects of clearing away heat and analgesia, scavenging free radicals and inhibiting lipid peroxidation damage. It is often used clinically to treat ulcerative colitis with remarkable curative effect.

The invention adopts the compound method of TNBS and ethanol to make the rat ulcerative colitis model. TNBS is a hapten substance. Ethanol dissolves the mucus on the surface of the intestinal mucosa, destroys the intestinal mucosal barrier, and makes TNBS become a complete antigen after combining with the macromolecular histone of the intestinal tissue, leading to an immune response against the intestinal mucosa and inducing ulcerative colitis. produce. Its pathological feature is that the colon tissue presents a chronic inflammatory change with a long course of disease, and the pathological changes are similar to human ulcerative colitis. Tests have confirmed that the gallic extract (high and low dose groups) has a definite therapeutic effect on colonic ulcers in rats induced by TNBS. The pathological results showed that there was no obvious abnormality in the colon tissues of the animals in the high-dose gallate extract group. In the model group, the colonic mucosa and submucosa were infiltrated with inflammatory cells, mainly neutrophils, and some epithelial shedding and mucosal necrosis occurred. Significant granulomas appeared. MPO is an enzyme present in neutrophils. It is an important indicator of neutrophil infiltration and an indicator of the degree of colitis damage. The experimental results show that the extract of gall seeds can significantly reduce the level of MPO in the serum of model rats. It shows that the gallic extract has obvious anti-inflammatory effect. Free radical damage is one of the important mechanisms of UC pathogenesis. SOD is an important enzyme for the body to scavenge oxygen free radicals. It plays a vital role in the balance of oxidation and anti-oxidation in the body. Oxidation reaction, and can stabilize the cell membrane, the experimental results show that the gallic extract significantly increased the level of SOD in the peripheral blood of rats, indicating that it has the ability to resist oxidative damage. This study shows that the effective parts of gallic seeds have significant therapeutic effects on ulcerative colitis, and the mechanism may be related to the anti-inflammatory and antioxidant effects of gallic seed extracts.

Example 6 Experimental Research on Anti-inflammatory, Analgesic and Antibacterial Treatment of Effective Fractions of Gallia in Preparation

Abstract: Objective: To observe the anti-inflammation, analgesic and antibacterial pharmacological effects of the effective part of gallaceae. Methods: Xylene-induced swelling of mouse ear shells, increased permeability of mouse abdominal capillaries induced by acetic acid, migration of leukocytes in mouse abdominal cavity induced by carrageenan, swelling of mouse paws induced by carrageenan, large The anti-inflammatory, analgesic and antibacterial effects of gallate extract were observed in rat cotton ball granuloma, writhing reaction of mice induced by acetic acid, and in vitro antibacterial of common intestinal bacteria. Results: Mice (2.5, 5.0, 10.0ml/kg) and rats (1.75, 3.5, 7.0ml/kg) were intragastrically administered the effective parts of gallaceae, which significantly inhibited the swelling of the mouse ear shell caused by xylene; antagonized the effect of acetic acid. Increased permeability of mouse abdominal capillaries induced by carrageenan; significantly reduced the migration of white blood cells in the peritoneal cavity of rats caused by carrageenan; significantly inhibited the swelling of mouse toes caused by carrageenan; inhibited the formation of cotton ball granuloma in rats Effect; can significantly reduce the number of writhing reactions in mice induced by acetic acid. In vitro antibacterial test against Salmonella typhi, Salmonella paratyphi B, Salmonella typhimurium, enterotoxigenic Escherichia coli, enteropathogenic Escherichia coli, enteroinvasive Escherichia coli, Shigella dysenteriae, Shigella flexneri The minimum bactericidal concentration (MBC) of 9 common indicator bacteria such as Bacillus genus and Shigella sonnei are 0.0650%, 0.0650%, 0.0650%, 0.0650%, 0.0650%, 0.150%, 0.150%, 0.150% respectively , 0.50%. Conclusion: The effective parts of Gallicia have obvious anti-inflammatory and analgesic effects, and have antibacterial effects on common pathogenic bacteria related to ulcerative colitis.

Gallaceae are the larvae of Cynips gallae-tinctoriceOliv, an insect of the family Apiaceae, which parasitize on the young branches of Quercus infectoriaOliv., a plant of the family Fagaceae, and produce galls. Gallatin is dry and cold in nature, bitter and astringent in taste, and has the functions of strengthening astringency, astringent, dampness, hemostasis, and anti-inflammatory. It is a commonly used medicinal material in Uighur medicine. The theory of Uyghur medicine believes that it can directly act on the intestinal wall to reduce the absorption and pinching force of the intestinal wall caused by colitis, astringe the intestines, reduce inflammation and diarrhea, open up intestinal resistance, and promote ulcer healing. Clinically used in the treatment of ulcerative colitis, the curative effect is remarkable. However, its mechanism of action in the treatment of ulcerative colitis has not yet been clarified. In this experiment, the anti-inflammatory, analgesic, and antibacterial effects of the effective parts of gallaceae were studied to explore its mechanism of action in the treatment of ulcerative colitis, which provides a basis for the development of new drugs for gallaceae. .

Experimental Materials and Instruments

Drugs and Reagents

Tested drug: the extract of the effective part of gallaceae (containing 40 mg of crude drug per milliliter), provided by the Uygur Medical Research Institute of Xinjiang Uygur Autonomous Region, batch number: 090926.

Test reagents: xylene, analytically pure, purchased from Tianjin North Chemical Glass Purchase and Sales Center (batch number 031228). Evansland: Provided by Shanghai Chemical Reagent Purchasing and Supply Station, imported by Fluka (batch number 82-11-02). Acetic acid: Zhengzhou Dezhong Chemical Reagent Factory (batch number 030506). Carrageenan: Provided by Pharmacology Department of Beijing Medical University. Indicator bacteria, Salmonella typhi (50071), Salmonella paratyphi B (50602), Salmonella typhimurium (50115), enterotoxigenic Escherichia coli (44815), enteropathogenic Escherichia coli (44336), enteroinvasive Escherichia coli (44850), Shigella dysenteriae (51570), Shigella flexneri (51142), Shigella sonnei (51592). Provided by the Department of Microbiology, Xinjiang Medical University.

Nutrient agar medium was produced by Hangzhou Tianhe Biological Reagent Co., Ltd. (batch number 200304). MH broth medium was provided by the Health and Defense Inspection Institute of the Logistics Department of Zhejiang Military Region (batch number 200304). MH agar medium, Hangzhou Tianhe Biological Reagent Co., Ltd. (batch number 200304). Normal saline: Shandong Hualu Pharmaceutical Co., Ltd. (batch number H20030128).

Test Instruments

BS110S electronic balance: Beijing Sartorius Balance Co., Ltd. DT200 electronic balance: Shanghai Hengping Scientific Instrument Co., Ltd. JM31000 electronic balance: Yuyao Jiming Weighing Calibration Equipment Co., Ltd. Outer diameter micrometer: Shanghai Measuring Tool & Cutting Tool Factory. Electronic thermometer: Fujian Kang Technology Industrial Co., Ltd. Stopwatch: Shanghai Jinxing Instrument Factory, QB/T1534-199. FM biochemical incubator: Shanghai Fuma Experimental Equipment Co., Ltd. YJ-875 ultra-clean workbench: Wujiang Hailiu Purification Equipment Co., Ltd. DM2500 Microscopic Imaging System: Product of Leica Company, Germany.

experimental animals

Wistar and SD male rats, normal grade; mice, Kunming species, normal grade, half male and half female. Provided by the Experimental Animal Center of Xinjiang Medical University, certificate of qualification: Xinjiang Yidongzi No. 16-003.

Method and Results

The effect of the effective part of gallacea on the xylene-induced swelling of the mouse ear shell 50 Kunming mice, weighing 20±2g, were divided into male and female. They were randomly divided into 5 groups with 10 rats in each group, which were blank control group, positive control group, small, medium and large dose groups of the effective parts of gallaceae. One hour after the medicine, 50 μl of xylene was applied on both sides of the right ear of the mice in each group. After one hour, the mice were sacrificed by dislocation of the neck. Mean t-test between groups, and the verification inhibition rate was calculated according to "(average swelling degree of blank control group-average swelling degree of drug-administered group)/average swelling degree of blank control group×100%".

Results: Gastrointestinal administration of different doses of the effective parts of gallaceae significantly inhibited the swelling of the ear shells of mice caused by xylene (results are shown in Table 1).

Table 1 Effects of effective parts of Galla on xylene-induced swelling of mouse ear shells

Note: Compared with blank control group, ***P<0.01, **P<0.05.

Effects of effective fractions of gallaceae on the increase in the permeability of the peritoneal capillaries in mice induced by acetic acid:

Take 50 Kunming mice, weighing 20±2g, half male and half male, and randomly divide them into 5 groups, 10 mice in each group, which are the blank control group, positive control group, small, medium and high dose groups of gallate extract. Once a day, continuous intragastric administration for 7 days. 1 hour after the last administration, inject 0.1ml/10g of 0.5% Evans blue normal saline solution into the tail vein of each mouse, and immediately ip 0.6% glacial acetic acid 0.2ml per mouse, kill the mice after 20 minutes, and wash the peritoneal cavity with 5ml normal saline , collect the washing solution, add physiological saline to the washing solution to 10ml, centrifuge at 3000rpm for 15min, take the supernatant and measure its OD value at a wavelength of 590nm by an ultraviolet spectrophotometer. The OD value represents the concentration of Evans blue in the flushing fluid, which indirectly reflects the degree of peritonitis exudation. Compared with the blank control group, the mean t test between groups was carried out to compare whether there was a significant difference. And the inhibition rate was calculated by "(average OD value of blank control group-average OD value of drug-administered group)/average OD value of blank control group×100%".

As a result, the OD value of the peritoneal washing liquid in each administration group of the effective part of gallaceae was significantly lower than that of the control group, indicating that the preparation significantly inhibited the increase in the permeability of the mouse peritoneal capillary caused by acetic acid (results are shown in Table 2).

Table 2 The effect of the effective part group of galla on the increase of the permeability of the mouse peritoneal capillary (X ± SD)

Note: Compared with blank control group, ***P<0.01.

Effects of the effective part of gallacea on the swelling induced by carrageenan in the paws of mice:

Take 50 Kunming mice, weighing 20±2g, half male and half male. They were randomly divided into 5 groups, 10 rats in each group, which were the blank control group, the positive control group, and the small, medium, and high-dose groups of gallate extract. Once a day, continuous intragastric administration for 7 days. One hour after the last administration, inject 0.06ml of 2% carrageenan under the skin of the central paw of the mouse, and measure the thickness of both paws with a micrometer at 1 h, 6 h, and 24 h after inflammation, so that the thickness of the paws before and after inflammation is the degree of swelling , for each group mean t-test.

The results showed that different doses of the effective part of Gallacea were administered intragastrically, and had an inhibitory effect on carrageenan-induced paw swelling in mice, and the effect could last for about 24 hours, especially in the high-dose group (see Table 3 for results).

Table 3 Effect of the effective part of gallacea on the swelling induced by carrageenan in the foot of mice (X±SD)

Note: Compared with blank control group, *P>0.05, **P<0.05, ***P<0.01.

The effect of the active part of gallat on the migration of white blood cells in the peritoneal cavity of rats:

Select 60 SD rats, weighing 200±20g, half male and half male. They were randomly divided into 6 groups, 10 rats in each group, which were the blank control group, model group, positive control group, small, medium and large dose groups of the effective parts of gallia. The rats in the blank control group and the model group were given normal saline by intragastric administration, the positive control group was intragastrically administered compound acetylsalicylic acid tablets, and the small, medium and large dose groups of the test substance were given corresponding doses of the effective part of gallate by intragastric administration, once a day , for 7 consecutive days. One hour after the last administration, rats in other groups except the control group were intraperitoneally injected with 2% carrageenan 0.4ml/rat. After 5 hours, the animals were sacrificed, and the peritoneal cavity was washed with 3ml of normal saline, and 1ml of washing liquid was collected and placed in a small In the test tube, the total number of white blood cells in each field of view was counted according to the white blood cell counting method, and the mean number of white blood cells was used as an index to conduct a t-test between groups.

The results showed that the ig administration of different doses of the effective part of gallacea had a significant inhibitory effect on the migration of leukocytes in the peritoneal cavity of rats induced by carrageenan (results are shown in Table 4).

Table 4 Effects of effective fractions of gallaceae on migration of rat peritoneal leukocytes

Note: ^△△△ P<0.01 compared with the blank control group; ***P<0.01 compared with the model group.

Effect of gallic seed extract on cotton ball granuloma in rats:

Take 40 Wistar rats, half male and half male, weighing 200±20g, and randomly divide them into 5 groups, 8 rats in each group. That is, the normal group, the 5-ASA group, and the three different doses of the effective parts of the gallic administration group. Under light ether anesthesia, after the anesthesia is successful, the lateral flanks of each rat are disinfected with iodine wine, and after deiodination with 75% alcohol cotton balls, each cut a small bite, and the subcutaneous tissue is expanded with vascular forceps. Sterilized cotton balls were implanted under the skin from the incision, and the skin was sutured. From the day after the operation, ig once a day for 7 consecutive days. On the 8th day, the animals were sacrificed, and the granulation tissue of the cotton balls was peeled off and taken out, dried in an oven at 60°C for 12 hours and weighed. Weight, minus the weight of the original cotton balls, is the net weight of granuloma, and converted into the mg number of granuloma in 100g body weight with the weight of rats, the average amount of granuloma in each group was used as the t test between groups, and "(blank control group) Net dry weight of granuloma - net dry weight of granuloma in the administration group)/net wet weight of granuloma in the blank control group × 100%" to calculate the inhibition rate.

The results showed that different doses of the effective parts of gallaceae were given by intragastric administration, and the formation of cotton ball granuloma in rats was obviously inhibited (results are shown in Table 5).

Table 5 Effects of effective fractions of gallaceae on cotton ball granuloma in rats

Note: Compared with blank control group, ***P<0.01.

Effects of effective fractions of gallacea on writhing response induced by acetic acid in mice:

Take 50 Kunming mice, weighing 20±2g, half male and half male. They were randomly divided into 5 groups, 10 rats in each group, which were blank control group, positive control group, small, medium and large dose groups of the effective parts of gallia. Once a day, continuous intragastric administration for 7 days. One hour after the last administration, each mouse was given 0.6% acetic acid 0.2ml/mouse ip, and the writhing response latency and the number of writhing times caused by pain within 15 minutes were observed for each mouse after ip acetic acid. Compared with the control group, the mean t test between groups was carried out to compare whether there was a significant difference. The analgesic percentage was calculated as "(average number of writhing times in the blank control group - average number of writhing times in the medication group)/average number of writhing times in the blank control group × 100%".

As a result, the effective part of Gallatin can significantly reduce the number of writhing reactions in mice caused by acetic acid, indicating that the preparation has a significant analgesic effect (results are shown in Table 6).

Table 6 Effects of the effective fractions of gallon on the writhing response of mice caused by acetic acid

Note: ***P<0.01 compared with blank control group.

Effects of effective fractions of Galla on nine common intestinal bacteria:

Working concentration of index bacteria: After 9 strains of index bacteria were recovered and passaged twice, under sterile normal saline, the working concentration was made up to about 1×10 ⁶ /ml by turbidimetric method. At the same time, they were diluted and inoculated on plates to calculate the number of viable bacteria. After the drug is filtered and sterilized, it is diluted with MH broth, and divided into small test tubes, each tube is 1.8ml, and the drug dilutions are 1:1, 1:2, 1:4, 1:8, 1:16 respectively. , 1:32, 1:64. Take 0.2ml of the above-mentioned bacterial working bacteria solution, add it to a test tube containing 1.8ml of drug broth, the final concentration of the indicator bacteria is about 1×10 ⁶ /ml, incubate at 37°C for 24 hours, inoculate plates to observe whether there is corresponding bacterial growth, and calculate Minimum bactericidal concentration (MBC), the test was repeated three times. Control group: medium aseptic control, drug aseptic control, and normal saline aseptic control.

The working concentration of indicator bacteria: 10 ⁷ cfu/ml; the working concentration of Salmonella typhi is 2.7×10 ⁷ cfu/ml, the final concentration of indicator bacteria is 2.7×10 ⁶ cfu/ml; the working concentration of Salmonella paratyphi B is 6.2×10 ⁷ cfu/ml, the concentration of indicator bacteria is 6.2×10 ⁶ cfu/ml; the working concentration of Salmonella typhimurium is 2.0×10 ⁷ cfu/ml, the concentration of indicator bacteria is 2.0×10 ⁶ cfu/ml; the working concentration of enterotoxigenic Escherichia coli is 0.6×10 ⁷ cfu/ml, index bacteria concentration 0.6×10 ⁶ cfu/ml; enteropathogenic E. coli working concentration 4.8×10 ⁷ cfu/ml, indicator bacteria concentration 4.8×10 ⁶ cfu/ml; enteroinvasive E. coli working concentration 0.7×10 ⁷ cfu/ml, indicator bacteria concentration 0.7×10 ⁶ cfu/ml; working concentration of Shigella dysenteriae 7.0×10 ⁷ cfu/ml, indicator bacteria concentration 7.0×10 ⁶ cfu/ml; Shigella flexneri The working ^{concentration} of Shigella sonnei is 1.8×10 ⁷ cfu/ml, and the index bacteria concentration is 1.8× ^{10 6 cfu} . /ml.

The results showed that the minimum bactericidal concentrations (MBC) of the nine index bacteria were 0.0650% for Salmonella typhi, 0.0650% for Salmonella paratyphi b, 0.0650% for Shigella dysenteriae, and 0.0650% for Shigella flexneri. Shigella sonnei 0.0650%, Salmonella typhimurium 0.150%, enterotoxigenic Escherichia coli 0.150%, enteroinvasive Escherichia coli 0.150%, enteropathogenic Escherichia coli 0.50%. Control: Aseptic growth in culture medium, aseptic growth in normal saline, and aseptic growth after disinfection of the three drugs (Table 7).

Table 7 Effects of effective fractions of gallia on nine common intestinal bacteria

discuss

Gallatum is a commonly used medicinal material in Uyghur medicine, and it is recorded in the Uyghur medicinal material standard-volume 1. It has the effects of astringent, dampness, hemostasis, anti-inflammatory and antiseptic. It is used for large intestine weakness, dysentery, habitual enteritis, sagging gums, periodontitis, bad breath, pharyngitis, etc. The Uygur Medical Hospital of Xinjiang Uygur Autonomous Region clinically used gallate extract to treat abdominal pain, diarrhea and sepsis caused by ulcerative colitis, and achieved remarkable curative effect.

In vivo experiments have shown that the administration of gallate extract to mice (2.5, 5.0, 10.0ml/kg) and rats (1.75, 3.5, 7.0ml/kg) can significantly inhibit the ear swelling of mice caused by xylene; Antagonize the increase of mouse abdominal capillary permeability caused by acetic acid; reduce the migration of white blood cells in the peritoneal cavity caused by carrageenan; significantly inhibit the swelling of mouse toes caused by carrageenan; affect rat cotton ball granuloma The formation has a significant inhibitory effect; it can significantly reduce the number of writhing reactions in mice induced by acetic acid. In vitro antibacterial studies have shown that the extracts of gallaceae are effective against Salmonella typhi, Salmonella paratyphi beta, Salmonella typhimurium, enterotoxigenic Escherichia coli, enteropathogenic Escherichia coli, enteroinvasive Escherichia coli, Shigella dysenteriae Genus, Shigella flexneri, Shigella sonnei and other 9 common indicator bacteria have inhibitory effect, and their minimum bactericidal concentration (MBC) is 0.0650% for Salmonella typhi and 0.0650% for Salmonella paratyphi B. %, Shigella dysenteriae 0.0650%, Shigella flexneri 0.0650%, Shigella sonnei 0.0650%, Salmonella typhimurium 0.150%, enterotoxigenic Escherichia coli 0.150%, intestinal invasion Enteropathogenic Escherichia coli 0.150% and enteropathogenic Escherichia coli 0.50%.

The above studies have shown that the extract of gall seeds has obvious anti-inflammatory and analgesic effects, and has antibacterial effects on common pathogenic bacteria related to ulcerative colitis.

Example 7

Clinical observation of 48 cases of anti-ulcerative colitis treated with the effective part of gallaceae:

Ulcerative colitis is a chronic nonspecific ulcerative inflammation. Lesions mainly occur in the large intestine mucosa and submucosa, mainly ulcers and erosions, with recurrent diarrhea, abdominal pain, and pus, blood and mucus as the main clinical manifestations. At present, the incidence of the disease is increasing, and its treatment is difficult. It is listed as one of the modern intractable diseases by the World Health Organization. The curative effect of western medicine is not satisfactory, and the adverse reaction is serious. Gallatin is dry and cold in nature, bitter and astringent in taste, and has the effects of strengthening astringency, astringent, dampness, hemostasis, and anti-inflammatory. It is a commonly used medicinal material in Uighur medicine, mainly used to treat ulcerative colitis. 48 cases of ulcerative colitis were treated with the effective parts of Gallatum, and the results are reported as follows:

data method

normal information

All 48 outpatients with mild and moderate ulcerative colitis were randomly divided into treatment group and control group. 24 cases in the treatment group, 16 males and 8 females; aged 20-65 years, average (42.4±12.5) years; disease duration 1-22 years, average (7.3±3.6) years; mild 11 cases, moderate 23 cases . 24 cases in the control group, 15 males and 9 females; aged 20-62 years, average (41.7±11.9) years: duration of disease 1-20 years, average (6.8±2.9) years: mild 12 cases, moderate 12 cases . After statistical analysis, there was no significant difference between the two groups in terms of gender, age, course of disease and disease severity (P>0.05), which were comparable.

Inclusion criteria

All cases had different degrees of diarrhea, abdominal pain and pus, blood and mucus in the stool, which conformed to the diagnostic criteria and curative effect evaluation criteria established by the 2000 Chengdu National Symposium on Inflammatory Bowel Disease. All cases are mild to moderate patients who meet this standard, where diarrhea > 6 times/d, severe mucus and bloody stool, body temperature above 37.5°C, pulse > 90 beats/min, hemoglobin < 100g/L, erythrocyte sedimentation rate > 30mm/ h severe patients were excluded.

treatment method

Treatment group: use 1.0 g of the effective part of gall, 4 times a day.

Control group: oral administration of sulfasalazine 1.0 g, 4 times a day; after the symptoms were relieved, it was changed to 1.0 g, 2 times a day. Both groups were treated for 8 weeks.

Observation indicators and methods

The improvement of symptoms and signs such as diarrhea, abdominal pain and pus and blood in the stool were observed before and after treatment; stool routine, blood routine, mucosal changes under colonoscopy, biopsy pathological examination; adverse reactions were observed.

Efficacy standard

The 1993 National Academic Symposium on Chronic Non-infectious Intestinal Diseases adopted the standards: namely, ①Short-term cure: clinical symptoms disappeared, and the mucosa was normal upon reexamination by colonoscopy. Withdrawal or only maintenance dose of drugs, observed for 6 months without recurrence. ②Effective: The clinical symptoms basically disappeared, and the mild inflammatory reaction of the mucosa and the formation of some pseudopolyps were reexamined by colonoscopy. ③ Ineffective: After treatment, the clinical symptoms, endoscopic and pathological examinations did not improve.

Statistical method

The ^χ2 test was performed with SPSS12.0 statistical software.

Observation results

The efficacy comparison is shown in Table 1.

Table 1 Comparison of clinical efficacy between the two groups (n=24)

group Recently cured efficient invalid Total effective rate (%) therapy group 16 (66.7%) 6 (25.0%) 2 (8.3%) 91.7 ^* control group 12 (50.0%) 7 (29.2%) 5 (20.8%) 79.2

^* : p<0.01, compared with the control group.

discuss

Ulcerative colitis is a colonic lesion syndrome characterized by chronic nonspecific inflammatory changes. The disease is easy to recur, protracted and difficult to heal, and has many complications. There are many theories about the etiology of ulcerative colitis, such as immune factors, genetic factors, mental factors, infection factors, and lack of protective substances. Recent studies have shown that the activity of superoxide dismutase (SOD) in the intestinal mucosa of patients with ulcerative colitis is low. Gallatin is dry and cold in nature, bitter and astringent in taste, and has the functions of strengthening astringency, astringent, dampness, hemostasis, and anti-inflammatory. It is a commonly used medicinal material in Uighur medicine. The theory of Uyghur medicine believes that it can directly act on the intestinal wall to reduce the absorption and pinching force of the intestinal wall caused by colitis, astringe the intestines, reduce inflammation and diarrhea, open up intestinal resistance, and promote ulcer healing. Clinically used in the treatment of ulcerative colitis, the curative effect is remarkable.

Observation results of clinical treatment show that the effective part of Gallatin has obvious therapeutic effect on ulcerative colitis, the total effective rate is 91.7%, the short-term cure rate is 66.7%, the effective rate is 25%, and the invalid rate is 8.3%, which is obviously better than the control group. Pharmacological studies have shown that the effective part of gallaceae has a significant therapeutic effect on rat ulcerative colitis; it can significantly increase the content of SOD in ulcer tissue (P<0.01), reduce the content of MPO (P<0.01), and has obvious anti-inflammatory, Analgesic and bacteriostatic against common pathogenic bacteria associated with ulcerative colitis. The effective part of gallaceae is a good medicine for treating ulcerative colitis.

Claims (1)

1. A use of the effective part of gallaceae in the preparation of a medicine for anti-ulcerative colitis, characterized in that the extraction method of the effective part of gallacea is carried out according to the following steps: a. Take the dried gallic medicinal material and crush it, pass through a No. 2 sieve, weigh 100g of medicinal material powder, add water to extract 3 times, add 5-15 times the amount of water each time, and extract for 20-120min; b. Concentrate the extract under reduced pressure to 500-1500ml, extract the concentrate with ether until the ether layer is clarified, and discard the ether layer; c. Extract the part of the water layer with ethyl acetate, recover the ethyl acetate, and dry it in vacuum at a temperature of 45-90°C to obtain 30-60% of the effective parts of the galls; d. Separating the chemical components of the active part of gallic acid obtained in step c by using a conventional gel chromatography column to obtain monomeric compounds, including gallic acid, meta-digallic acid, methyl gallate, 1,2, 3,6-tetra-O-galloyl-β-D-glucose, 1,2,3,4,6-penta-O-galloyl-β-D-glucose.