CN103948754A - Drug for treating renal-deficiency humid-heat type chronic nephritis - Google Patents

Abstract

The invention discloses a medicine for treating damp-heat chronic nephritis due to kidney deficiency, which is prepared by extracting raw materials according to the following proportions by weight: 20-40 parts of astragalus; 8-12 parts of Atractylodes macrocephala; 8-12 parts of cornus 10-30 parts of Eucommia; 10-20 parts of Alisma; 10-30 parts of Shiwei; 10-30 parts of Hedyotis diffusa; It has good therapeutic function and is safe to take, and is suitable for the prevention and treatment of chronic nephritis. The invention also discloses its preparation method and application.

Description

A kind of medicine for treating chronic nephritis with damp-heat due to kidney deficiency and preparation method thereof

technical field

The invention belongs to the technical field of traditional Chinese medicine compositions, and more specifically relates to a traditional Chinese medicine formula medicine for treating chronic nephritis and its preparation method and application.

Background technique

Chronic nephritis is a common kidney disease with a high prevalence rate and a high mortality rate, and its incidence rate is increasing year by year. Chronic nephritis is caused by many reasons, and the pathological changes are different. Its main manifestations are edema, proteinuria, hematuria, high blood pressure and decreased renal function. In the ancient books of traditional Chinese medicine, there is no such disease name as "chronic glomerulonephritis". , but according to its clinical manifestations and onset characteristics, the disease belongs to the categories of "edema", "lumbar pain", "kidney wind", "diarrhea", "off grid", "kidney fatigue", "constipation", "constipation" in traditional Chinese medicine Addition", "hematuria" and other categories. The location of the disease is mainly in the spleen and kidney. Traditional Chinese medicine believes that the spleen controls the movement and transformation, clears the turbid secretion, completes the absorption and transportation of the essence of water and grain, and participates in the water metabolism of the body; Kidney governs storage, storing Yuan Yin and Yuan Yang; Kidney governs water and completes water metabolism. If pathological damage occurs, the spleen and kidney will be deficient, the spleen will lose its transport and transformation, and the essence cannot be absorbed and transported normally; the kidney will lose its seal, and the essence will leak into the urine, resulting in proteinuria; Blood enters the urine from the outside of the pulse, resulting in hematuria; the spleen fails to transform, and the water dampness stops inside, overflowing the skin; the kidney element is insufficient, the bladder loses gasification, the opening and closing is difficult, and the water dampness stops inside, resulting in edema. Clinical observation shows that the pathogenic factors of this disease are mainly damp-heat, blood stasis, turbid toxin, and water-dampness. Therefore, the pathogenesis of this disease is deficiency in origin and excess in superficiality, and deficiency and excess are mixed. Kidney disease is not only quite common, but also the medical expenses are very high. The annual dialysis fee for each patient with uremia is 50,000-60,000 RMB. It is the first cause of uremia dialysis in my country, so it is of great significance to actively treat chronic nephritis.

At present, western medicine and western medicine mainly use hormones, immunosuppressants, ACEI/ARB drugs, etc. for the treatment of nephritis. The market is badly in need of a traditional Chinese medicine preparation for treating kidney deficiency and damp-heat type chronic nephritis with a wide range of applications and little side effects.

Contents of the invention

The object of the present invention is to provide a Chinese medicinal preparation for treating chronic nephritis of kidney deficiency-damp-heat type with remarkable curative effect and little side effects and a preparation method thereof.

The present invention is achieved through the following technical solutions:

A medicine for treating chronic nephritis of kidney deficiency and damp-heat type, which is prepared by extracting raw materials in the following proportions by weight: 20-40 parts of astragalus; 8-12 parts of Atractylodes macrocephala; 8-12 parts of cornus; 10- 30 parts; Alisma 10-20 parts; Shiwei 10-30 parts; Hedyotis diffusa 10-30 parts; Panax notoginseng 3-7 parts.

The above-mentioned drug for treating chronic nephritis of kidney deficiency and damp-heat type is prepared by extracting raw materials in the following proportions by weight: 30 parts of astragalus; 10 parts of Atractylodes macrocephala; 10 parts of cornus; 20 parts of Eucommia; 20 parts; Hedyotis diffusa 20 parts; Panax notoginseng 5 parts.

The preparation method of the above-mentioned medicine for treating chronic nephritis of damp-heat type due to kidney deficiency comprises the following steps:

a) Take Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes Rhizome and Hedyotis diffusa according to the above parts by weight, extract volatile oil by steam distillation, and clathrate with β-cyclodextrin;

b) Take Astragalus, Eucommia, Alisma, Shiwei, and Notoginseng five flavors according to the above parts by weight, extract through ethanol reflux, filter, recover ethanol, and concentrate to obtain an alcohol extraction concentrate;

c) combining the medicinal residues after the steam distillation method in step a, the medicinal residues after alcohol extraction in step b, and Cornus officinalis, decocting in water, filtering, concentrating, cooling, and adding ethanol for alcohol precipitation to obtain a concentrated solution of water extraction and alcohol precipitation; The water-extracted alcohol-precipitated concentrated solution is combined with the alcohol-extracted concentrated solution in step b, and spray-dried to make a dry extract powder;

d) Take the volatile oil clathrate of step a and the dry extract powder of step c, perform wet granulation, and dry to obtain the drug.

The above-mentioned medicine for treating chronic nephritis of kidney deficiency and damp-heat type, the steam distillation method in step a of the preparation method, uses 6 times the weight of the medicinal material to soak in water for 1 hour, distills for 8 hours to extract volatile oil, adds 6 times the weight of β-cyclodextrin to the volatile oil, and uses Saturated aqueous solution method 40°C electric stirring, the stirring speed is 120r/min, after stirring for 1.5 hours, the clathrate is carried out, and the clathrate is vacuum-dried at 40°C.

The above-mentioned drug for treating kidney deficiency and damp-heat chronic nephritis is extracted by ethanol reflux in step b of the preparation method. The amount of alcohol added is 12 times the total weight of the decoction pieces, and the volume concentration of ethanol is 70%. The reflux extraction is divided into three times, each time for 2 hours, and the extracts are combined , filtered, and the filtrate recovered ethanol under reduced pressure, and concentrated until it had no alcohol smell.

The above-mentioned drug for treating chronic nephritis of kidney deficiency and damp-heat type, in step c of the preparation method, is decocted with water, and the amount of water added is 12 times the total weight of the decoction pieces, decocted twice, each time for 2 hours, combined with water decoction, filtered , when the filtrate is concentrated to 50°C, the relative density is 1.05-1.08, cool, add ethanol until the alcohol content does not exceed 50%, stir well, let stand for 48 hours, filter, recover ethanol from the filtrate, combine with the above-mentioned alcohol extraction concentrate and concentrate to 50% The relative density at ℃ is 1.10-1.15, adding appropriate amount of dextrin, spraying and drying into dry extract powder under the condition of inlet air temperature of 165-175℃ and outlet air temperature of 85-90℃.

The above-mentioned drug for treating chronic nephritis of kidney deficiency and damp-heat type, in the preparation method step d, take the volatile oil clathrate in step a and the dry extract powder in step c, add appropriate amount of sugar powder and dextrin, mix evenly, carry out wet granulation, dry, The ready-to-obtain medicine, the dosage form is tablet or granule or hard capsule.

Application of the above-mentioned medicine for treating chronic nephritis of kidney deficiency-damp-heat type in the preparation of medicine for treating kidney-deficiency damp-heat type chronic nephritis.

In the present invention, Atractylodes macrocephala is the dry rhizome of Atractylodes macrocephala Koidz. of Compositae plant, Astragalus is the dry root of leguminous plant Astragalus membranaceus (Fisch.) Bge.Var.mongholicus (Bge.) Hsiao, and Cornus officinalis is the plant Cornus officinalis of Cornaceae Comus officinalis Sieb.et Zucc. Dry mature pulp, Eucommia is the dry bark of Eucommia ulmoides Oliv., and Alisma is the dry tuber of Alisma orientalis (Sam.) Juzep. Araliaceae plant Panax notoginseng (Burk.) F.H.Chen's dry root, Shi Wei is the water keel family plant Shiwei Pyrrosialingua (Thunb.) Farwell's dry leaves, diffusa is the Rubiaceae plant Hedyotis diffusaWilld . of dried whole herb.

Compared with the existing traditional Chinese medicine for treating chronic nephritis, the present invention has the following advantages:

(1) Astragalus membranaceus and Cornus officinalis in the prescription are monarch drugs. Raw Astragalus membranaceus is sweet in taste, slightly warm in nature, beneficial to qi and yang, invigorating the spleen and diuresis, and benefiting the health and strengthening the exterior. Modern pharmacological studies have confirmed that it has a strong immune regulation effect, can promote the differentiation and maturation of T lymphocytes, induce cells to produce interferon, and at the same time can relieve vasospasm, improve microcirculation, enhance tissue hypoxia resistance, reduce proteinuria, Diuretic and other effects. Cornus cornus is sweet, slightly bitter, enters the liver and kidney meridians, has the functions of nourishing the liver and kidney, astringent essence, and has the function of protecting and increasing the preservation of kidney units. Chronic nephritis, diabetic nephropathy, chronic pyelonephritis and other common chronic kidney diseases are characterized by deficiency of spleen and kidney qi and deficiency of both qi and yin. Astragalus membranaceus and Cornus officinalis in the prescription can invigorate qi and invigorate the spleen, and the other can nourish yin and invigorate the kidney, which together achieve the functions of nourishing qi and nourishing yin and nourishing the spleen and kidney, both of which are monarch drugs. In the prescription, Atractylodes macrocephala and Eucommia are ministerial medicines. Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes Rhizome is sweet, bitter, and warm in nature. It has the functions of invigorating qi, invigorating the spleen, drying dampness and diuresis. Because of edema, it is still a common symptom of chronic nephritis, and the disease of edema "is rooted in the kidney" and "is controlled in the spleen". Atractylodes macrocephala It can not only help Astragalus to invigorate Qi, but also remove dampness and transport the spleen to prevent Astragalus from obstructing Qi. Eucommia ulmoides is sweet in taste, slightly pungent, warm in nature, enters the liver and kidney meridians, has the function of nourishing the liver and kidney, and strengthening the muscles and bones. Tonifying kidney qi is a ministerial drug. In the prescription, Alisma, Shiwei, Hedyotis diffusa, and Panax notoginseng are adjuvant drugs. Alisma is sweet and bland in taste, cold in nature, can diuretic and diuretic, expel heat and relieve stranguria. It is mainly used for the treatment of adverse bladder gasification, water-damp retention, difficulty in urination, and fullness of edema. Pharmacological studies have proved that Alisma alisma can lower blood pressure, blood sugar, blood fat, increase urine output, excretion of urea and sodium chloride. Shi Wei is sweet and bitter in taste, slightly cold in nature, can diuresis for treating stranguria, clears away heat and stops bleeding. It is often used for hot stranguria, bloody stranguria, hematuria and other diseases with damp heat in the lower Jiao. Pharmacological studies have shown that Shiwei has anti-inflammatory and anti-pathogenic microorganisms and diuretic effects. It can be used clinically to treat acute and chronic nephritis and pyelonephritis. In addition to eliminating edema, it can also reduce proteinuria and improve kidney function. Hedyotis diffusa is sweet and light in taste, cool in nature, can clear away heat and detoxify, promote blood circulation and detumescence. Modern pharmacological studies have found that it has a strong anti-pathogenic microorganism effect, can stimulate the proliferation of the reticuloendothelial system, increase the phagocytic ability of phagocytes, and enhance the defense ability of the body. In patients with chronic nephritis, damp-heat in the lung and kidney meridians is the common cause of relapse or aggravation of the disease. Although Hedyotis diffusa has the function of clearing away heat and detoxifying, its nature and taste are sweet and mild without the risk of bitter cold to hurt the stomach. Since dampness and heat are common in chronic kidney disease syndromes, and dampness and heat are an important factor for the recurrence or aggravation of various chronic kidney diseases, clearing dampness and heat is a common treatment principle for chronic kidney disease. In the prescription, Alisma, Shiwei, and Hedyotis diffusa are used together to clear heat and detoxify, dampness and swelling. Radix Notoginseng is sweet and slightly bitter in taste, warm in nature, and its raw product function is to dissipate blood stasis and stop bleeding, reduce swelling and relieve pain, and stop bleeding without leaving blood stasis. For a variety of bleeding syndromes and bruises, swelling and pain. Modern pharmacological studies have found that it can increase thrombin in the blood and constrict local blood vessels, thereby achieving hemostasis. The total saponins of Panax notoginseng can inhibit platelet aggregation, and have anticoagulant activity in vitro. It can be seen that the two-way regulating effect of notoginseng on promoting coagulation and anticoagulation is consistent with traditional Chinese medicine's view that its effect of promoting blood circulation and stopping bleeding is consistent. Hematuria is common clinically in chronic kidney disease, and the most common manifestation of renal pathological changes is the proliferation of various renal intrinsic cells, often accompanied by glomerulosclerosis and renal interstitial fibrosis. The effect of blood stasis is suitable for a variety of chronic kidney diseases. The invention has better advantages in improving clinical symptoms of patients, protecting renal function and safety. Traditional Chinese medicine believes that the main clinical manifestations and test indicators of chronic nephritis are often related to kidney deficiency and damp heat, which belongs to the syndrome of deficiency of the origin and excess of the standard. The deficiency of spleen and kidney qi is the root, and the dampness, heat and stasis of toxin are the standard. Clearing away damp-heat, mainly treating chronic glomerulonephritis spleen-kidney qi-yin deficiency, damp-heat accumulation syndrome.

(2) Most of the traditional Chinese medicines are decoctions at present, with large prescriptions, many medicinal flavors, and inconvenient use, but the present invention has a better protective function for chronic nephritis;

(3) The preparation process is simple, the extraction efficiency is high, and the cost is saved;

(4) The medicine of the present invention can be made into dosage forms such as granules, capsules, tablets or pills, which is convenient to take;

(5) The results of clinical and pharmacological tests of the present invention show that the Chinese medicine preparation can significantly improve clinical syndromes, reduce urinary protein, lower blood creatinine and blood urea nitrogen levels, inhibit inflammatory reactions and release of inflammatory mediators, and promote the development of glomerular pathological damage. Therefore, it has a good therapeutic effect on chronic nephritis, and it has been developed into a health care product with auxiliary protective function for chronic nephritis or a new drug for treating chronic nephritis. The market capacity is large, and the economic and social benefits are obvious.

Detailed ways

Example 1:

Take Astragalus 200g; Atractylodes macrocephala 120g; Cornus 80g Eucommia 300g; Alisma 100g;

a. Take Atractylodes macrocephala and Hedyotis diffusa to extract the volatile oil by steam distillation, soak in water with 6 times the weight of the medicinal material for 1 hour, distill for 8 hours to extract the volatile oil, add 6 times the weight of β-cyclodextrin to the volatile oil, and use a saturated aqueous solution Method 40 ℃ electric stirring, stirring speed of 120r/min, stirring for 1.5 hours after clathrate, clathrate 40 ℃ vacuum drying.

b. Take Astragalus, Eucommia, Alisma, Shiwei, Sanqiwuwei, and extract through ethanol reflux. The amount of alcohol added is 12 times the total weight of the decoction pieces, and the volume concentration of ethanol is 70%. Reflux extraction in three times, each time 2 hours, combined the extracts, filtered, and the filtrate recovered ethanol under reduced pressure, and concentrated until it had no alcohol smell. Filtrate, recover ethanol, concentrate to obtain alcohol extraction concentrate;

c. Merge the medicinal residues after the steam distillation in step a, the medicinal residues after the alcohol extraction in step b, and Cornus officinalis, decoct with water, add water 12 times the total weight of the decoction pieces, and decoct twice, each time 2 hours, combined decoction, filtered, the relative density of the filtrate was 1.05 when the filtrate was concentrated to 50°C, cooled, added ethanol until the alcohol content was 50%, stirred well, left for 48 hours, filtered, the filtrate recovered ethanol, and the above step b When the alcohol extraction concentrates are combined and concentrated to 50°C, the relative density is 1.15, and an appropriate amount of dextrin is added, and spray-dried under the conditions of an inlet air temperature of 165°C and an outlet air temperature of 90°C to obtain a dry extract powder.

d. Take the volatile oil clathrate in step a and the dry extract powder in step c, add appropriate amount of dextrin and mix to carry out wet granulation, dry, compress into tablets, and prepare into tablets.

Example 2:

Take Astragalus 400g; Atractylodes macrocephala 80g; Cornus 120g Eucommia 100g; Alisma 200g; Shiwei 100g; Hedyotis diffusa 300g;

a. Take Atractylodes macrocephala and Hedyotis diffusa to extract the volatile oil by steam distillation, soak in water with 6 times the weight of the medicinal material for 1 hour, distill for 8 hours to extract the volatile oil, add 6 times the weight of β-cyclodextrin to the volatile oil, and use a saturated aqueous solution Method 40 ℃ electric stirring, stirring speed of 120r/min, stirring for 1.5 hours after clathrate, clathrate 40 ℃ vacuum drying.

b. Take Astragalus, Eucommia, Alisma, Shiwei, Sanqiwuwei, and extract through ethanol reflux. The amount of alcohol added is 12 times the total weight of the decoction pieces, and the volume concentration of ethanol is 70%. Reflux extraction in three times, each time 2 hours, combined the extracts, filtered, and the filtrate recovered ethanol under reduced pressure, and concentrated until it had no alcohol smell. Filtrate, recover ethanol, concentrate to obtain alcohol extraction concentrate;

c. Merge the medicinal residues after the steam distillation in step a, the medicinal residues after the alcohol extraction in step b, and Cornus officinalis, decoct with water, add water 12 times the total weight of the decoction pieces, and decoct twice, each time 2 hours, combined the water decoction, filtered, the relative density of the filtrate was 1.08 when concentrated to 50°C, cooled, added ethanol until the alcohol content was 50%, stirred well, left for 48 hours, filtered, the filtrate recovered ethanol, and the above step b When the alcohol extraction concentrates are combined and concentrated to 50°C, the relative density is 1.10, an appropriate amount of dextrin is added, and the air temperature at the inlet is 175°C, and the air temperature at the outlet is 85°C, and it is spray-dried into a dry extract powder.

d. Take the volatile oil clathrate in step a and the dry extract powder in step c, add an appropriate amount of dextrin, mix well, carry out wet granulation, and dry to prepare granules.

Example 3:

Take Astragalus 300g; Atractylodes macrocephala 100g; Cornus 100g, Eucommia 200g; Alisma 150g; Shiwei 200g; Hedyotis diffusa 200g;

a. Take Atractylodes macrocephala and Hedyotis diffusa to extract the volatile oil by steam distillation, soak in water with 6 times the weight of the medicinal material for 1 hour, distill for 8 hours to extract the volatile oil, add 6 times the weight of β-cyclodextrin to the volatile oil, and use a saturated aqueous solution Method 40 ℃ electric stirring, stirring speed of 120r/min, stirring for 1.5 hours after clathrate, clathrate 40 ℃ vacuum drying.

b. Take Astragalus, Eucommia, Alisma, Shiwei, Sanqiwuwei, and extract through ethanol reflux. The amount of alcohol added is 12 times the total weight of the decoction pieces, and the volume concentration of ethanol is 70%. Reflux extraction in three times, each time 2 hours, combined the extracts, filtered, and the filtrate recovered ethanol under reduced pressure, and concentrated until it had no alcohol smell. Filtrate, recover ethanol, concentrate to obtain alcohol extraction concentrate;

c. Merge the medicinal residues after the steam distillation in step a, the medicinal residues after the alcohol extraction in step b, and Cornus officinalis, decoct with water, add water 12 times the total weight of the decoction pieces, and decoct twice, each time 2 hours, combined the water decoction, filtered, the relative density of the filtrate was 1.06 when concentrated to 50°C, cooled, added ethanol until the alcohol content was 50%, stirred well, left for 48 hours, filtered, the filtrate recovered ethanol, and the above step b When the alcohol extraction concentrates are combined and concentrated to 50°C, the relative density is 1.13, an appropriate amount of dextrin is added, and the air temperature at the inlet is 170°C, and the outlet air temperature is 87°C, and it is spray-dried into a dry extract powder.

d. Take the inclusion compound of volatile oil in step a and the dry extract powder in step c, add appropriate amount of dextrin and mix evenly, carry out wet granulation, dry, fill capsules, and prepare hard capsules.

Embodiment 4: Pharmacological experiment of the present invention (Yishen Qingli granule):

1 Materials and methods

1.1. Sample: prepare the present invention according to above-mentioned embodiment 2, obtain dry extract powder.

1.2. Experimental animals: Wistar rats, body weight (150-200) g, half male and half female, provided by the Experimental Animal Center of Nanjing University of Traditional Chinese Medicine, experimental animal production license number: SCXK (Su) 2008-0010, animals are randomized according to body weight Divide into 5 groups, 10 in each group.

1.3. Main instruments and reagents: automatic biochemical analyzer; biochemical kits were purchased from Nanjing Jiancheng Institute of Bioengineering; 30% ethanol was prepared with distilled water; liver tissue pathology sections were made by the Department of Pathology, Jiangsu Provincial Hospital of Traditional Chinese Medicine.

2. The effect of Yishen Qingli granules of the present invention on rat Heymann's glomerulonephritis

2.1 Preparation of Renal Cortex Freund's Complete Adjuvant Suspension

Take 150-200g Wistar rats, kill them by devertebral dissection, quickly extract the kidneys and insert them into catheters, wash them repeatedly with normal saline until they turn white, cut the kidneys, take 3g of renal cortex to make a homogenate, mix it with Freund's complete adjuvant to 40ml, add Physiological saline 40ml, thoroughly emulsified, set aside.

2.2 Heymann glomerulonephritis model

Select 150-200g Wistar rats, feed and observe them for 1w, put them into metabolic cages to collect 24h urine, and use a urine analyzer to detect urine protein. Rats with negative urine protein test were used for modeling. Among them, 8 rats were used as normal control group. The remaining rats were intraperitoneally injected with renal cortex Freund's complete adjuvant suspension 2ml/rat. The same method and the same dose were injected once every 2w for a total of 5 injections. 1w after the last injection, the rats were placed in rat metabolic cages, and the urine was collected for 24 hours, and the urine protein content was measured by the biuret method. A significant increase in the amount of urinary protein was defined as a successful indicator of the model. Select 50 model rats and divide them into 5 groups at random, namely model group, positive drug group, and test drug (high, medium, low) group, with 10 rats in each group. Rats in each group were given intragastric administration at the 9th week of the experiment, administered once a day for 28 days in a row. Shenqingli granule extract, the administration volume is 1mL/100g.

2.3 Observation indicators

2.3.124h urine output and urine protein detection: after the modeling (before administration), the rats were placed in the rat metabolic cage to collect 24h urine output on the 14th and 28th day of administration, and the urine protein was determined by the biuret method content, see Table 1 for the results.

2.3.2 Detection of BUN, GRE, serum total protein, and serum albumin: Blood was collected from the orbit after modeling (before administration), on the 14th day of administration, and at the end of administration (28 days after administration) for blood biochemistry The results are shown in Table 2 and Table 3.

2.4 Experimental results

2.4.1 General situation After the formal immunization, the immunized animals have different degrees of lethargy, loose and messy hair, weak grasping, loose stools, and ascites. After administration, the animals in the model group were listless, and the loose and dull hair gradually increased. The Shenyan Kangfu tablet group and the Yishen Qingli granule group improved compared with each other, but the spirit was still listless, and the hair was loose and dull.

2.4.2 Dynamic changes of urinary protein Urine was collected in metabolic cages of rats, and the amount of urinary protein was measured for 24 hours.

Table 124-hour urine protein quantification (mg/24 hours, n=10)

group Before administration Dosing for 14 days 28 days blank group 1.6076±0.8448 2.993±1.46 3.1±0.55 model group 4.244±1.283 ^** 7.219±2.65 ^* 6.94±2.40 ^** Nephritis rehabilitation tablet group 4.061±1.357 ^** 4.66± ^0.45 * 3.78±1.22 ^△ high dose group 4.909±1.952 ^** 5.32±0.99 ^* 3.50±1.46 ^△ Middle dose group 4.721±1.597 ^** 5.04±1.11 ^* 4.78±1.29 ^* low dose group 5.059±1.985 ^** 5.42±1.11 ^* 4.38±0.909 ^*

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01; compared with the model group ^{, △} P<0.05, ^△△ P<0.01.

The results showed that the 24-hour urine protein of the rats increased significantly after modeling ( ^** P<0.01), and there was no significant difference among the modeling groups. After 14 days of treatment, the effects of each administration group were not significant (compared with the blank group, ^* P<0.05). After 28 days of treatment, the Shenyan Kangfu Tablets group and the high-dose Yishen Qingli Granules group had significant effects (compared with the model group ^△ P<0.05), while the effects of the Yishen Qingli Granules medium and low dose groups were not significant (compared with the blank group, ^* P<0.05).

2.4.3 Dynamic changes in serum albumin Blood was collected from the orbit of rats to determine serum albumin.

Table 2 Serum albumin (g/L, n=10)

group Before administration Dosing for 14 days 28 days blank group 33.3±2.23 32.68±1.41 32.08±2.29 model group 28.98±2.065 ^** 26.98±2.55 ^** 26.08±2.05 ^** Nephritis rehabilitation tablet group 28.13±2.89 ^** 24.94±2.94 ^** 27.78±0.85 ^** high dose group 28.38±3.20 ^** 26.58±2.14 ^** 29.98±1.19 ^△ Middle dose group 28.61±2.67 ^** 25.92±3.12 ^** 31.38±0.97 ^△△ low dose group 28.48±1.86 ^** 24.82±3.59 ^** 27.72±2.61 ^*

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01; compared with the model group ^{, △} P<0.05, ^△△ P<0.01.

The results showed that the serum albumin of the rats increased significantly after modeling (compared with the blank group, ^** P<0.01), and there was no significant difference among the modeling groups. After 14 days of treatment, the effects of each administration group were not significant (compared with the blank group, ^** P<0.01). After 28 days of treatment, the high and medium dose groups of Yishen Qingli Granules had a significant effect (compared with the model group ^△ P<0.05, ^△△ P<0.01). Shenyan Kangfu Tablets group and Yishen Qingli Granules low-dose group had no significant effect (compared with blank group, ^* P<0.05, ^** P<0.01).

2.4.4 Dynamic changes in serum creatinine Blood was collected from the orbits of rats to determine serum creatinine.

Table 3 Serum creatinine (umol/L, n=10)

group Before administration Dosing for 14 days 28 days blank group 20.33±1.86 22.8311.47 23.40±1.81 model group 30.40±4.92 ^** 30.14±3.97 ^** 27.66±1.36 ^* Nephritis rehabilitation tablet group 31.80±5.71 ^** 32.42±4.72 ^** 27.611.67 ^* high dose group 29.40±5.54 ^** 28.25±2.06 ^** 23.67±2.32 ^△△ Middle dose group 32.2±1.92 ^** 28.7515.67 ^* 24.71±2.42 ^△ low dose group 31.2±8.34 ^* 31.25±4.36 ^** 28.2±1.48

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01; compared with the model group ^{, △} P<0.05, ^△△ P<0.01.

The results showed that the serum creatinine of the rats increased significantly after modeling (compared with the blank group, ^* P<0.05, ^** P<0.01), and there was no significant difference among the modeling groups. After 14 days of treatment, the effects of each administration group were not significant (compared with the blank group, ^** P<0.01). After 28 days of treatment, the high and medium dose groups of Yishen Qingli Granules had a significant effect (compared with the model group ^△ P<0.05, ^△△ P<0.01). The Shenyan Kangfu tablet group and the low-dose Yishen Qingli granule group had no significant effect (compared with the blank group, ^* P<0.05).

From the above results, it can be seen that Yishen Qingli Granules significantly reduced 24-hour urine protein and creatinine, and increased serum albumin. Shenyan Kangfu Tablets significantly reduced 24-hour urine protein and decreased the tendency of serum creatinine rise, but the effect was not significant. For albumin There was no significant improvement in the reduction. Compared with the two drugs, Yishen Qingli Granules are better than Shenyan Kangfu Tablets in reducing serum muscle levels and increasing serum albumin.

Therefore, Yishen Qingli granules have a good therapeutic effect on Heymann nephritis, can improve the symptoms of Heymann nephritis rats, reduce the 24-hour urine protein quantity and serum creatinine level and increase serum albumin.

3. The effect of Yishen Qingli Granules on C-BSA glomerulonephritis in rats

3.1 Experimental method

3.1.1 Preparation of cationized bovine serum albumin (C-BSA)

(1) Take 67mL of anhydrous ethylenediamine (EDA) solution, add 500mL of double distilled water, add 350ml of 6mol/L hydrochloric acid to adjust the pH value to 4.75, cool to 25°C, stir, add natural bovine serum albumin (N -BSA) 5g solution dissolved in 25ml double distilled water, after stirring, add 1.8g carbonized diimide hydrochloride (EDCHCL) to maintain 25°C, keep stirring for 2h, keep the pH value at 4.75, and finally add 4mol/L (PH value 4.75 ) 30 ml of acetic acid buffer solution to terminate the reaction, and obtain a C-BSA solution (PI=9.2) with an increased isoelectric point.

(2) Place the C-BSA solution in a semi-permeable bag, dialyze with double distilled water at 4°C for 72 hours (change the water once every 3-5 hours), and freeze-dry to obtain dry BSA powder. Store at -30°C for later use.

(3) Take the above powder, that is, cationized bovine serum albumin, dissolve it with 0.01mol/LPH7.4PBS before use, and filter to sterilize.

3.1.2 Preparation of incomplete Freund's adjuvant

10g of lanolin, 40mL of liquid paraffin, sterilized at 108°C for 15 minutes; irradiated with a mortar and ultraviolet lamp for 2 hours, added lanolin into the mortar on an ultra-clean workbench, added liquid paraffin while grinding according to the mass-volume ratio of lanolin at 1:3, and ground To uniformity, that is, incomplete Freund's adjuvant. Store in the refrigerator (4°C) for later use.

3.1.3 C-BSA nephritis model

Take 72 rats, after adaptive feeding for 3 days, collect 24h urine, measure 24h urine protein by biuret colorimetry, and randomly divide them into normal group, positive drug control group, model group according to 24h urine protein amount and body weight, There are six groups of Yishen Qingli Granules with high, medium and low doses. For pre-immunization, 1 mg of C-BSA was dissolved in 0.5 mL of PBS, fully emulsified with the same amount of IFA, and then subcutaneously injected into rats at multiple points; for formal immunization, starting from the 8th day, C-BSA 16 mg/kg was injected into the tail vein every other day. A total of 29d. 24-hour urine was collected and 24-hour urine protein was tested. Rats with significantly increased urinary protein in each group began to be administered by intragastric administration at the 12th week of the experiment, once a day for 28 days, the blank group and the model group were given normal saline, and the positive drug group was given Shenyan Kangfu tablet suspension liquid, and the test drug group was given Yishen Qingli granule extract. The administration volume was 1mL/100g.

3.1.4 IL-6 determination method and establishment of standard curve

3.1.4.1 IL-6 determination method

1) Take out the kit from the refrigerator 20 minutes in advance to equilibrate the room temperature. Take out the strips required for the test from the sealed bag, put the unused strips and desiccant back into the pot bag and seal them at 4°C, add the specimens and standards of different concentrations (0pg/ml well plus reagent diluent) into the corresponding wells . Seal the reaction well with sealing tape and incubate for 40 min in a 37°C incubator.

Note: IL-6 standard was diluted to 2000, 1000, 500, 250, 125, 62.5, 31.25 (pg/ml) in sequence.

2) Wash the plate 6 times, add 50ul each of distilled water and primary antibody working solution (except the blank). Seal the reaction wells with sealing tape and incubate in a 37°C incubator for 20 min.

3) Wash the plate 6 times, and add enzyme-labeled antibody working solution (100ul/well). Seal the reaction well with adhesive tape and incubate at 37°C for 10 min.

4) Wash the plate 6 times (dilute the concentrated washing solution with double distilled water 1:20), add 100ul/well of the substrate working solution, and incubate for 15min in the dark at 37°C in a dark place in an incubator.

5) Add stop solution 100ul/well, measure A value immediately after mixing.

3.1.4.2 Establishment of IL-6 standard curve

Taking the concentration of the standard product as the abscissa and the A value as the ordinate, draw the IL-6 standard curve, and calculate the IL-6 level in the supernatant of each group according to the standard curve.

3.2 Observation indicators

3.2.124h urine output and urine protein detection: respectively after molding (before administration), the rats were put into the rat metabolic cage to collect 24h urine output on the 14th and 28th day of administration, and the urine protein was measured by biuret method content.

3.2.2 Detection of BUN, GRE, serum total protein, and serum albumin: blood was collected from the orbit after modeling (before administration), on the 14th day of administration and at the end of administration (28 days after administration) for blood biochemistry detection.

3.2.3 Detection of serum IL-6: blood was collected from the orbit after film formation (before administration) and at the end of administration (28 days after administration), and was completed by double-antibody sandwich ELISA using BIO-RAD680 microplate reader IL-6 detection.

3.3 Experimental results

3.3.1 General situation After the formal immunization, the immunized rats showed varying degrees of lethargy, loose and messy hair, weak grasping, loose stools, and ascites. After administration, the rats in the model group were listless, with loose and dull hair, weak grasping, and loose stools, etc., gradually aggravated. The Shenyan Kangfu tablet group and the Yishen Qingliji group improved compared with each other, but the rats still showed listlessness and hair loss. Loose and dull, unable to grasp, loose and thin stool.

3.3.2 Dynamic changes of urinary protein Urine was collected in metabolic cages of rats, and the amount of urinary protein was measured for 24 hours. The results are shown in Table 4.

Table 4 24-hour urine protein quantification (mg/kg·24 hours, n=12)

group Before administration Dosing for 14 days 28 days blank group 32.4±5.24 16.58±6.54 12.2±3.35

model group 66.8±29.95 ^* 33.61±9.45 ^** 32.45±10.67 ^** Nephritis rehabilitation tablet group 101.7±32.9 ^** 21.62±7.93 ^△ 15.22±6.20 ^△ high dose group 57.2±19.73 ^* 18.13±8.95 ^△ 16.76±3.65 ^△△ Middle dose group 74.9±25.77 ^** 22.18±5.10 ^△ 14.78±5.31 ^△ low dose group 51.56±12.08 ^* 24.81±4.88 ^* 15.97±3.66 ^△△

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01; compared with the model group ^{, △} P<0.05, ^△△ P<0.01.

The results showed that the 24-hour urine protein of the rats after modeling was significantly higher than that of the normal group ( ^* P<0.05, ^** P<0.01), and there was no significant difference among the modeling groups. After 14 days of treatment, the effects of Yishen Qingli Granules in high and medium dose groups and Shenyan Kangfu Tablets group were significant (compared with the model group, ^△ P<0.05), while the effect of the low-dose group was not significant (compared with the blank group, ^* P<0.05). At 28 days, the effects of each group were significant (compared with the model group, ^△△ P<0.05, ^△△ P<0.01), and the effects of the low and high dose groups were extremely significant (compared with the model group, ^△△ P<0.01). This medicine can significantly reduce urine protein.

3.3.3 Dynamic changes in creatinine Blood was collected from the orbits of rats to determine serum creatinine. The results are shown in Table 5.

Table 5 Serum creatinine (umol/L, n=12)

group Before administration Dosing for 14 days 28 days blank group 16.57±1.84 24±1 23.8±0.75 model group 22.88±2.26 ^* 27±1.94 ^** 26.2±1.17 ^** Nephritis rehabilitation tablet group 24.33±3.77 ^* 24.33±1.37 ^△ 23.17±2.19 ^△ high dose group 24.77±5.55 ^* 24.33±1.22 ^△△ 23.8±0.98 ^△ Middle dose group 25.67±4.67 ^** 23.89±2.81 ^△ 24.2±0.75 ^△ low dose group 27.2±5.56 ^* 24.29±1.98 ^△ 24±1.41 ^△

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01; compared with the model group ^{, △} P<0.05, ^△△ P<0.01.

The results showed that the serum creatinine of the rats increased significantly after modeling (compared with the blank group, ^* P<0.05, ^** P<0.01), and there was no significant difference among the modeling groups. After 14 days of treatment, the effects of each administration group were significant (compared with the model group ^{, △△} P<0.05, ^△△ P<0.01), and the effect of the high-dose group was more significant (compared with the model group, ^△△ P<0.01). After 28 days of treatment, the effects of each group were significant ( ^△ P<0.05). This medicine can significantly lower blood creatinine.

3.3.4 Dynamic changes in serum albumin Blood was collected from the orbit of rats to determine serum albumin. The results are shown in Table 6.

Table 6 Serum Albumin (g/L, n=12)

group Before administration Dosing for 14 days 28 days blank group 35.93±0.81 37.3±1.08 35.83±0.75 model group 34.29±1.30 ^* 35.57±0.57 ^** 34.62±0.16 ^** Nephritis rehabilitation tablet group 33.4±1.15 ^** 35.6±1.26 ^** 35.24±0.38 ^△

high dose group 32.7±3.57 ^* 36.96±1.37 ^△ 35.8±0.94 ^△ Middle dose group 34.23±1.53 ^* 37.26±0.90 ^△△ 35.76±0.96 ^△ low dose group 33.56±1.82 ^* 35.99±0.78 ^* 32.13±3.07 ^*

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01; compared with the model group ^{, △} P<0.05, ^△△ P<0.01.

The results showed that the serum albumin of the rats increased significantly after modeling (compared with the blank group, ^* P<0.05 ^** , P<0.01), and there was no significant difference among the modeling groups. After 14 days of treatment, the high and medium dose groups of Yishen Qingli Granules had a significant effect (compared with the model group ^△ P<0.05, ^△△ P<0.01). After 28 days of treatment, the Shenyan Kangfu tablet group, Yishen Qingli granule high-dose and medium-dose groups all had significant effects (compared with the model group ^△ P<0.05). The drug can significantly increase serum albumin, and the effect is earlier than that of Nephritis Rehabilitation Tablets.

3.3.5 Dynamic changes in cholesterol Blood was collected from the orbits of rats to measure serum cholesterol. The results are shown in Table 7.

Table 7 serum cholesterol (mmol/L, n=12)

group Before administration Dosing for 14 days 28 days blank group 1.78±0.13 1.47±0.11 1.35±0.07 model group 2.36±0.42 ^* 1.78±0.19 ^** 1.61±0.19 ^* Nephritis rehabilitation tablet group 2.7±0.42 ^* 1.82±0.29 ^* 1.53±0.12 ^* high dose group 2.16±0.41 ^* 1.84±0.32 ^* 1.36±0.13 ^△ Middle dose group 2.13±0.31 ^* 1.55±0.09 ^△ 1.40±0.11 ^△ low dose group 2.26±0.38 ^* 1.72±0.20 ^* 1.30±0.18 ^△

Note: Compared with the blank group, ^* P<0.05, ^** P<0.01; compared with the model group ^{, △} P<0.05, ^△△ P<0.01.

After modeling, blood cholesterol in each group was significantly increased (compared with the normal group, ^* P<0.05), and there was no significant difference among the modeling groups. After 14 days of treatment, the middle dose group of Yishen Qingli Granules had a significant effect (compared with the model group, ^△ P<0.05). At 28 days, the effect of each dosage group of Yishen Qingli granule was significant (compared with the model group, ^△ P<0.05). It is suggested that the drug can improve the rising trend of cholesterol, and the effect is earlier and better than that of Nephritis Rehabilitation Tablets.

3.3.6 Comparison of serum IL-6 levels in rats before and after treatment Take blood from the orbits of rats to measure serum IL-6. The results are shown in Table 8.

The comparison (pg/mL, )

group no Before administration 28 days blank group 9 14.09±3.65 19.43±4.05 model group 8 105.97±44.17 ^** 257.00±81.49 ^** Nephritis rehabilitation tablet group 8 305.52±195.99 ^** 140.28±102.34 ^△# high dose group 9 262.26±112.10 ^** 54.56±34.18 ^△## low dose group 8 199.03±56.04 ^** 59.95±40.63 ^△#

Note: Compared with blank group, ^* P<0.05, ^** P<0.01; compared with model group, ^△ P<0.05, ^△△ P<0.01; compared with before treatment, ^# P<0.05, ^## P<0.01 .

After modeling, the serum IL-6 levels of rats in each group were significantly increased (compared with the normal group, P<0.01), and there was no significant difference among the modeling groups. On day 28, the levels of serum IL-6 in rats in the Shenyan Kangfu tablet group, Yishen Qingli granule high-dose and low-dose groups were all lower than those in the model group, and there was a significant difference between them (P<0.05).

Therefore, the low-dose and high-dose groups of Yishen Qingli Granules can significantly reduce 24-h urine protein ( ^△△ P<0.01). The effect of high dose group can significantly reduce creatinine ( ^△△ P<0.01) and serum IL-6 level. The Shenyan Kangfu Tablet group and the high-dose and middle-dose groups all significantly decreased serum albumin ( ^△ P<0.05), which was earlier than that of the Shenyan Kangfu Tablet. Each dose group significantly improved the trend of increasing cholesterol ( ^△ P<0.05), and its effect was earlier and better than that of Renyan Kangfu Tablet.

This study confirmed that Yishen Qingli granule has a good therapeutic effect on C-BSA nephritis, can improve its symptoms, reduce 24h urine protein quantification and serum creatinine level, increase serum albumin and improve the trend of cholesterol elevation, among which The effect was particularly evident in the high-dose group.

Claims (8)

1. a medicine that is used for the treatment of the damp-heat type chronic nephritis of suffering from a deficiency of the kidney, is characterized in that, the crude drug of pressing row weight portion proportioning extracts and is prepared from: Radix Astragali 20-40 part; Rhizoma Atractylodis Macrocephalae 8-12 part; Fructus Corni 8-12 part; Cortex Eucommiae 10-30 part; Rhizoma Alismatis 10-20 part; Folium Pyrrosiae 10-30 part; Herba Hedyotidis Diffusae 10-30 part; Radix Notoginseng 3-7 part.

2. be used for the treatment of as claimed in claim 1 the damp-heat type chronic nephritis medicine of suffering from a deficiency of the kidney, it is characterized in that, the crude drug of pressing row weight portion proportioning extracts and is prepared from: 30 parts of the Radixs Astragali; 10 parts of the Rhizoma Atractylodis Macrocephalaes; 10 parts of Fructus Corni; 20 parts of the Cortexs Eucommiae; 15 parts of Rhizoma Alismatis; 20 parts of Folium Pyrrosiae; 20 parts of Herba Hedyotidis Diffusaes; 5 parts of Radix Notoginseng.

3. be used for the treatment of as claimed in claim 1 the damp-heat type chronic nephritis medicine of suffering from a deficiency of the kidney, it is characterized in that preparation method comprises the following steps:

A) by above-mentioned parts by weight, get the Rhizoma Atractylodis Macrocephalae, Herba Hedyotidis Diffusae two tastes are through extraction by steam distillation volatile oil, with beta-cyclodextrin inclusion compound;

B) by above-mentioned parts by weight, get the Radix Astragali, the Cortex Eucommiae, Rhizoma Alismatis, Folium Pyrrosiae, the Radix Notoginseng five tastes, through alcohol reflux, filter, and reclaim ethanol, concentrate to obtain alcohol extraction concentrated solution;

C) medicinal residues after the medicinal residues after step a steam distillation, b alcohol extraction and Fructus Corni are merged, through decocting, boil, filter, concentrated, cooling, add ethanol precipitate with ethanol and obtain water extract-alcohol precipitation concentrated solution; The alcohol extraction concentrated solution of water extract-alcohol precipitation concentrated solution and step b is merged, and spraying is dry, makes dry extract;

D) get the volatile oil clathrate compound of step a and the dry extract of c carries out wet granulation, dry, obtain medicine.

4. the damp-heat type chronic nephritis medicine of suffering from a deficiency of the kidney that is used for the treatment of as claimed in claim 3, it is characterized in that steam distillation in preparation method step a, use 6 times of water gagings of medical material weight to soak 1 hour, distill and within 8 hours, extract volatile oil, volatile oil adds 6 times of weight beta-schardinger dextrin-s, adopts 40 ℃ of electric stirrings of saturated water solution method, and mixing speed is 120r/min, stir and carry out enclose after 1.5 hours, 40 ℃ of vacuum dryings of clathrate.

5. the damp-heat type chronic nephritis medicine of suffering from a deficiency of the kidney that is used for the treatment of as claimed in claim 3, it is characterized in that alcohol reflux in preparation method step b, institute's alcohol adding amount is 12 times of decoction pieces gross weight, ethanol volumetric concentration is 70%, minute three reflux, extract,, each 2 hours, merge extractive liquid,, filter, decompression filtrate recycling ethanol, is concentrated into without alcohol taste.

6. the damp-heat type chronic nephritis medicine of suffering from a deficiency of the kidney that is used for the treatment of as claimed in claim 3, it is characterized in that in preparation method step c, through decocting, boil, amount of water is 12 times of decoction pieces gross weight, divide secondary to decoct, each 2 hours, merge decocting liquid, filter, when filtrate is concentrated into 50 ℃, relative density is 1.05～1.08, cooling, add ethanol to alcohol content and be no more than 50%, stir evenly, place 48 hours, filter, filtrate recycling ethanol, while being concentrated into 50 ℃ with above-mentioned alcohol extraction concentrated solution merging, relative density is 1.10～1.15, add dextrin appropriate, in import pathogenic wind-warm, it is 165～175 ℃, under 85～90 ℃ of conditions of outlet pathogenic wind-warm, be spray dried to dry extract.

7. the damp-heat type chronic nephritis medicine of suffering from a deficiency of the kidney that is used for the treatment of as claimed in claim 3, it is characterized in that in preparation method steps d, get and in step a volatile oil clathrate compound and step c, get that dry extract adds appropriate Icing Sugar, dextrin mixes and carries out wet granulation, dry, obtain medicine, dosage form is tablet or granule or hard capsule.

8. be used for the treatment of as claimed in claim 1 the application that the damp-heat type chronic nephritis medicine of suffering from a deficiency of the kidney is suffered from a deficiency of the kidney in damp-heat type chronic nephritis medicine in preparation treatment.