CN114470101A - Traditional Chinese medicine composition and preparation method thereof, and oral preparation containing the same - Google Patents

Abstract

The invention discloses a traditional Chinese medicine composition, a preparation method thereof, and an oral preparation containing the same. The mass ratio of each raw material in the traditional Chinese medicinal composition based on raw meal is as follows: see serum 1000: cassia seed 100-2000: hawthorn 10-1000 : Lycium barbarum 5-500; the traditional Chinese medicine composition provided by the invention has high purity of active ingredients, mild action process, quick effect and little side effect.

Description

Traditional Chinese medicine composition and preparation method thereof, and oral preparation containing the same

technical field

The invention belongs to the field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition, a preparation method thereof, and an oral preparation containing the same.

Background technique

See Serum Liparis nervosa (Thunb) Lindl. It belongs to the Orchidaceae Sheep's ear allium, a terrestrial herb, also known as "rock taro", "black blue", "chumpty child", "sheep ear garlic", etc. It is widely distributed in It is a traditional folk medicinal plant in my country, such as Yunnan, Guizhou, Sichuan, Guangdong and Guangxi. Hemostasis, etc.

At present, domestic and foreign researches on serotonin mainly focus on the extraction and analysis of its chemical components and the pharmacological activities of some active components in anti-tumor, antibacterial, anti-inflammatory, anti-oxidative, and hemostatic effects; , Clinical treatment of hypolipidemic.

High blood pressure is a very common disease nowadays, most of which occur in middle-aged and elderly people. However, with the accelerated pace of life and increased pressure, many young people also suffer from high blood pressure. High blood pressure is a common heart disease in internal medicine. Vascular disease, which has many causes, is essentially both genetic and environmental.

Hyperlipidemia is generally concentrated in elderly patients. The elderly's blood lipids are higher than the normal prescribed range, and it is easy to form such diseases. Hyperlipidemia includes hypercholesterolemia, hypertriglyceridemia and complex hyperlipidemia ; High blood lipids can easily lead to high blood pressure, high blood lipids will cause the human body to form atherosclerosis, lead to myocardial dysfunction, and also lead to coronary heart disease. When the human body forms atherosclerosis due to long-term high blood lipids, it will cause coronary artery disease. The blood flow decreases, the vascular cavity narrows, and the amount of blood injected into the myocardium will also decrease, resulting in myocardial ischemia, angina pectoris, and coronary heart disease; high blood lipids can lead to liver damage, and long-term high blood lipids can lead to fatty liver , hepatic atherosclerosis is damaged, and the hepatic lobules are also damaged, resulting in structural changes and damage to liver function; hyperlipidemia is easy to form vascular sclerosis, after the formation of atherosclerosis, a large amount of lipids and proteins in the plasma It is deposited in the medium, reducing the speed of blood flow, and after oxidation, it becomes rancid and deposited on the endothelium of the arterial tube. It will adhere to the blood vessel wall for a long time, which will damage the arterial blood vessel skin and cause vascular sclerosis.

At present, the main treatment methods for hypertension and hyperlipidemia include exercise therapy, diet therapy, traditional Chinese medicine conditioning, and drug therapy. Antihypertensive drugs are effective, but most of them have certain side effects, which can damage the heart, kidneys and other organs to a certain extent.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a traditional Chinese medicine composition and an oral preparation containing the traditional Chinese medicine composition, which can effectively reduce blood pressure and blood lipids of human body, and have mild action process and few side effects.

The object of the present invention is also to provide a preparation method of a traditional Chinese medicine composition, which can effectively extract the active ingredients in the serum—total alkaloids and anthraquinones in cassia seeds, hawthorn and wolfberry, and reduce the ineffective ingredients. and irritating ingredients, so that the action process of the traditional Chinese medicine composition is milder, the side effects are reduced, and the blood pressure lowering and blood lipid lowering effects are improved.

The technical scheme adopted in the present invention is that the preparation method of the traditional Chinese medicine composition specifically comprises the following steps:

In step 1, the dried serum is washed and sieved after removing impurities, repeatedly extracted with an acidic polar solution, the combined extracts are neutralized with a dilute alkaline solution to make the system pH=6.5-7.5, and the polar solvent is recovered in vacuum to obtain an extract;

Step 2, dispersing the extract with water, adding ethanol for precipitation for 24 hours, filtering, recovering ethanol from the filtrate, extracting the alcohol extract with chloroform, and separating and purifying the extracted components to obtain the serum extract;

Step 3, extract the cassia seed extract, the hawthorn extract and the wolfberry extract respectively, and mix the serum extract with each of the extracts to obtain a traditional Chinese medicine composition.

Further, the specific process of repeatedly extracting in the step 1 is as follows:

Use an acidic polar solvent for extraction at 25°C to 90°C for 1 to 10 hours, and then use an acidic polar solvent for extraction at 25°C to 90°C for 0.5 to 1.5 hours. The pH of the acidic polar solvent is 1.0 to 3.0.

Further, in the described step 2, the separation and purification process is as follows:

The silica gel column was used for chromatography, and CHCl ₃ /MeOH was used as the elution system for gradient elution, and the active compounds were collected by fractions, which was the serum extract.

Further, the process of extracting the cassia seed extract is as follows:

The cassia seeds were dried and powdered, wetted with 15% sulfuric acid solution for 5 days, then extracted with chloroform under reflux for 3 hours, allowed to cool for a while, and filtered with suction to separate the acid water layer in the extract, and repeatedly washed with distilled water to obtain the chloroform extract;

Use pH=8.0 buffer to repeatedly extract the chloroform extract to obtain the raffinate phase, and repeatedly extract the raffinate phase with 0.2% NaOH solution;

Mix the NaOH extract, add hydrochloric acid dropwise under stirring until the pH of the system is 3, after the precipitation is completely separated out, suction filter, use distilled water to wash the precipitate to neutrality, and dry;

Use acetone to dissolve the precipitate, use silica gel column for chromatography, use petroleum ether-ethyl acetate to elute, collect yellow bands, combine fractions, concentrate, stand for crystallization, collect crystals, repeat column chromatography separation, use acetone for recrystallization , to obtain cassia seed extract.

Further, the extraction process of the hawthorn extract and the wolfberry extract is as follows:

Soak hawthorn or wolfberry in distilled water for 30min, heat to boiling and keep slightly boiling for 60min, add distilled water to hawthorn or wolfberry and decoct again for 30min after pressure filtration, combine the decoction liquid, concentrate in 50℃ water bath and slowly concentrate to obtain hawthorn extract or wolfberry extract.

For the traditional Chinese medicine composition, the quality ratio of each raw material based on raw meal is as follows: see serum 1000: cassia seed 100-2000: hawthorn 10-1000: wolfberry 5-500.

An oral preparation containing a traditional Chinese medicine composition, the oral preparation is a tablet or a capsule.

Further, it is prepared by the following mass ratios of various substances: Chinese medicine composition 10: dextrin 3: sodium carboxymethyl cellulose 1: sodium hydroxypropyl cellulose 2: magnesium stearate 0.2.

Further, the oral preparation is a capsule, which is prepared by the following mass ratios of various substances: traditional Chinese medicine composition 10: dextrin 5: sodium carboxymethyl cellulose 1: magnesium stearate 0.2.

The beneficial effects of the present invention are as follows: in the embodiment of the present invention, extracts of cassia seed, medlar, and hawthorn are used, which are processed into tablets or made into capsules, which can be used for the treatment of hypertension and hyperlipidemia. Fast effect, short course of treatment, and less toxic and side effects.

Description of drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the following briefly introduces the accompanying drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only These are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can also be obtained according to these drawings without creative efforts.

Fig. 1 is a graph showing the effect of traditional Chinese medicine composition 1 on pancreatic lipase activity.

Figure 2 is a graph showing the effect of oleic acid on HepG2 cells.

Fig. 3 is the effect diagram of the effect of traditional Chinese medicine composition on triglyceride synthesis.

Figure 4 is a graph showing the effect of traditional Chinese medicine compositions on lipid synthesis.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

The quality ratio of each raw material for the traditional Chinese medicine composition for lowering blood pressure and blood lipid in this application is as follows: see Serum 1000: Cassia 100-2000: Hawthorn 10-1000: Lycium barbarum 5-500, the quality of each raw material is based on raw materials count.

Example 1

The preparation process of the traditional Chinese medicine composition is as follows:

Step 1, take 1000 g of dry serum, wash and remove impurities, crush and pass through a 20-mesh sieve, and use an acidic polar solvent with a volume of 20 times the mass of the serum with pH=1.0-3.0 to extract 1- 10h, extract with 10 times more acidic polar solvent under the same conditions for 0.5-1.5h, combine the extracts, neutralize the system with dilute alkali solution to make the pH=6.5-7.5, and recover the polar solvent in vacuum to obtain the extract;

Use an appropriate amount of water to disperse the extract, add ethanol to the volume concentration of 80% to 90%, stir evenly, precipitate for 24 hours, filter, recover ethanol from the filtrate, use chloroform and ethyl acetate to extract the filtered alcohol extract, and then The extracted components are separated and purified to obtain a series of extracts;

The specific separation and purification process is as follows:

Chloroform part was chromatographed with silica gel column, gradient elution was carried out with CHCl ₃ /MeOH as elution system, and the active compound Liparis alkaloid (total alkaloid) was obtained by fractional collection, including extract 1, extract 2 and extract 3;

The ethyl acetate part was chromatographed on a silica gel column, and the gradient elution was carried out with CHCl ₃ /MeOH as the elution system, and the active compounds Phenolic glycosides (phenolic glycosides) were obtained by fractional collection, including extract 4, extract 5 and extract 6 ;

The active compound Liparis alkaloid and the active compound Phenolic glycosides were concentrated in vacuum to dryness, respectively, and dispersed with a small amount of purified water to obtain extract I and extract II.

Step 2, extract cassia seed extract, hawthorn extract, wolfberry extract respectively;

Step 3: Mixing and compounding the extract I, the extract of Mingzi, the hawthorn extract, and the extract of Lycium barbarum to obtain the traditional Chinese medicine composition 1; thing 2.

When the extraction time in the step 1 is short, the effective substances in the serum have not been completely dissolved, the yield of the extract is low, and the serum resources will be wasted. When the extraction time is long, the dissolution of the effective substances has already reached equilibrium. , prolonging the time will only increase the cost of manpower and material resources, and at the same time, the leaching amount of impurities in the extract will also increase, reducing the extraction effect.

At the same time, when the temperature of polar solution extraction is low, it is not conducive to the dissolution of effective substances. When the temperature is high, it may destroy the effective components in the serum and affect the quality of the serum extract.

In step 1, the volume concentration of ethanol when ethanol is used for extraction is too small to achieve the purpose of removing most of the alcohol-insoluble impurities. If the volume concentration is too high, the consumption of ethanol will be increased, resulting in waste; the precipitation time is too short, so that the impurities in the serum are not visible. Sufficient sedimentation, incomplete removal of impurities, and excessive time, resulting in waste of equipment and manpower.

The extraction process of cassia seed extract is as follows:

Dry and powder the Cassia seeds, weigh 200g and place it in a 1000mL~20000mL round-bottom flask or extraction tank, add 200mL~4000mL of sulfuric acid solution with a volume concentration of 15% to wet for 5 days, then add 500mL~10000mL of chloroform to reflux for extraction for 3 hours, and let it sit for a while. After cooling, suction filtration, discard the residue, transfer the chloroform extract into a separating funnel/separator, and after separating the acid water layer, repeatedly wash the chloroform extract 3 times with 75 mL to 1500 mL of distilled water to obtain the chloroform extract.

Place the chloroform extract in a 100mL~20000mL separatory funnel/separator, repeat the extraction with 150~3000mL pH=8.0 buffer for 5 times to obtain a raffinate phase, and place the raffinate phase in a 1000mL~20000mL separatory funnel In the / separator, repeatedly extract 4 times with 200mL-4000mL volume concentration of 0.2% NaOH.

Mix the sodium hydroxide extract and place it in a 1000mL~20000mL beaker/reactor, add hydrochloric acid dropwise with stirring to pH=3.0, after the precipitation is complete, filter with suction, wash with a small amount of distilled water to be neutral, and the precipitate is naturally dried spare.

Weigh 1 g of the precipitate and place it in an evaporating dish, add an appropriate amount of acetone to dissolve, add 25 g of silica gel for column chromatography, stir evenly, and evaporate to dryness in a fume hood for later use.

Weigh 200 g of silica gel plus petroleum ether-ethyl acetate (28:1), stir evenly, put it in a chromatography column, dry sample, elute with petroleum ether-ethyl acetate, collect yellow color bands, each 20 mL, Check each fraction with a silica gel G plate, combine the fractions, concentrate a little, stand for crystallization, collect the crystals, repeat the separation by column chromatography again, and recrystallize with analytically pure acetone to obtain the cassia extract.

The preparation process of hawthorn extract is as follows:

Take 100g of hawthorn and put it in a beaker, add an appropriate amount of distilled water to immerse the medicinal materials, soak for 30 minutes, heat to boiling, keep slightly boiling for 60 minutes, filter by pressure, add 5 times its mass of distilled water to the medicinal residues and decocted again for 30 minutes according to the law, combine and decocted for 2 times. The solution was vacuum concentrated and then placed in a 50°C water bath to slowly evaporate and concentrate to obtain the hawthorn extract, which was stored in a 4°C refrigerator for later use.

The preparation process of wolfberry extract is as follows:

Take 50g of wolfberry and put it in a beaker, add an appropriate amount of distilled water to immerse the medicinal materials, soak for 30 minutes, heat to boiling, keep a slight boil for 60 minutes, filter, add 4 times its mass of distilled water to the medicinal residues and cook again for 30 minutes according to law, and combine the 2 decoctions , vacuum concentrated and then placed in a 50°C water bath to slowly evaporate and concentrate to obtain Lycium barbarum extract, which was stored in a 4°C refrigerator for later use.

Example 2

Using the preparation process described in Example 1 to prepare a traditional Chinese medicine composition, the proportions of the raw materials are as follows: See serum 1000: Cassia 100: Hawthorn 10: Lycium barbarum 5, and the blood-seeing extract in the traditional Chinese medicine composition is the chloroform part isolated total alkaloids.

Example 3

Using the preparation process described in Example 1 to prepare a traditional Chinese medicine composition, the proportions of the raw materials are as follows: See serum 1000: Cassia seed 400: Hawthorn 200: Lycium barbarum 100, the See Blood Extract in the traditional Chinese medicine composition is the chloroform part isolated total alkaloids.

Example 4

Using the preparation process described in Example 1 to prepare a traditional Chinese medicine composition, the proportions of the raw materials are as follows: See serum 1000: Cassia seed 800: Hawthorn 400: Lycium barbarum 200, the See Blood Extract in the traditional Chinese medicine composition is the chloroform part isolated total alkaloids.

Example 5

Using the preparation process described in Example 1 to prepare a traditional Chinese medicine composition, the proportions of the raw materials are as follows: see serum 1000: cassia 1200: hawthorn 600: wolfberry 300, the see blood extract in the traditional Chinese medicine composition is the chloroform part isolated total alkaloids.

Example 6

Using the preparation process described in Example 1 to prepare a traditional Chinese medicine composition, the proportions of the raw materials are as follows: See serum 1000: Cassia seed 1600: Hawthorn 800: Lycium barbarum 400, the See Blood Extract in the traditional Chinese medicine composition is the chloroform part isolated total alkaloids.

Example 7

Using the preparation process described in Example 1 to prepare a traditional Chinese medicine composition, the proportions of the raw materials are as follows: See serum 1000: Cassia 2000: Hawthorn 1000: Lycium barbarum 500, and the see blood extract in the traditional Chinese medicine composition is the chloroform part isolated total alkaloids.

Example 8

80 experimental mice were selected, and 10 of them were divided into eight groups, wherein two groups were blank groups, three groups were experimental groups of serum extracts, and three groups were Chinese medicine composition 1 (prepared in Example 1) Experimental group, see serum extract experimental group and traditional Chinese medicine composition 1 experimental group were divided into low-dose group, middle-dose group and high-dose group;

One, adaptive feeding

The mice were fed environmentally adaptive feeding for 3-5 days in a standard laboratory environment, and then weighed, the tail tip blood was collected, and the serum was collected by centrifugation. ), low-density lipoprotein cholesterol (LDLC), and high-density lipoprotein cholesterol (HDLC).

2. Experiment

The blank group 1 was fed with basal feed and drinking water, and was gavaged with normal saline, and the amount of normal saline was 0.3 mL/10 g based on the weight of the mice.

The blank group 2 was fed with high-fat feed and drinking water. The high-fat feed was mixed with the following substances according to the following mass percentages: basal feed 75%, protein powder 15%, lard 10%, gavage with normal saline, physiological The dosage of saline is 0.3mL/10g according to the mass of mice.

See serum extract experimental groups were fed with high-fat feed and drinking water. The extract was administered by gavage, and the dosage of serum extract was 0.3mL/10g according to the mass of mice; the concentration of serum extract used in the low-dose group was 20 μg/mL, the concentration of serum extract in the middle-dose group was 50 μg/mL, and the concentration of serum extract used in the middle-dose group was 50 μg/mL. The concentration of the serum extract in the dosage group was 100 μg/mL, and the serum extract was the unseparated and purified extract prepared in Example 1.

Traditional Chinese medicine composition 1 experimental group was fed with high-fat feed and drinking water. Compound 1 was administered by gavage, and the dosage of traditional Chinese medicine composition 1 was 0.3 mL/10g according to the mass of mice; the concentration of traditional Chinese medicine composition 1 used in the low-dose group was 20 μg/mL, the concentration of traditional Chinese medicine composition 1 in the middle-dose group was 50 μg/mL, and the concentration of traditional Chinese medicine composition 1 in the low-dose group was 50 μg/mL. The concentration of traditional Chinese medicine composition 1 in the dosage group was 100 μg/mL.

Raised for 28 days according to the above method, weighed once every 3 days, fasted without water the day before the experiment, weighed 2 hours after the last administration, blood was collected from the heart, and serum was collected by centrifugation. Esters (TG) content, low density lipoprotein cholesterol (LDLC) content and high density lipoprotein cholesterol (HDLC) content.

It can be seen from blank group 1 and blank group 2 that feeding mice with high-fat diet can significantly increase their blood pressure and blood lipids. Obvious, and the mice showed symptoms such as loss of appetite, weight loss, diarrhea, and with the increase in the concentration of serum extracts, the blood pressure and blood lipid lowering effects were slightly improved, but the side effects were also more obvious; use traditional Chinese medicine composition 1 to treat After mice, the blood pressure and blood lipids of the mice were significantly reduced, and the effect was proportional to the concentration of the traditional Chinese medicine composition 1. Very few mice showed symptoms of decreased appetite, but had little effect on body weight and no diarrhea.

Example 9

1. Determination of hypolipidemic activity

The blood lipid-lowering activity of the traditional Chinese medicine composition was determined by measuring the triglyceride and cholesterol contents in HepG2 cells in vitro.

(1) Cell culture

HepG2 cells were placed in high-glucose DMEM medium containing 10% fetal bovine serum, 100 μg/mL penicillin, and 100 μg/mL streptavidin at 37 °C in a cell incubator with 5% _CO . white.

(2) Influence experiment of Chinese medicine composition 1 and Chinese medicine composition 2 prepared in Example 1 on the viability of HepG2 cells

First pre-treat HepG2 cells:

After HepG2 cells were trypsinized, 3×10 ⁵ cells/well were seeded into 96-well plates, and incubated at 37°C for 24 h. After the incubation, the cell supernatant was removed, and the cells containing different concentrations of Chinese medicine composition 1 were used. , Chinese medicine composition 2. The growth medium containing different molar concentrations of oleic acid was incubated at 37°C for 24h.

The mass concentrations of Chinese medicine composition 1 and Chinese medicine composition 2 are 0, 10 μg/mL, 50 μg/mL, 100 μg/mL, 200 μg/mL, 500 μg/mL, respectively, and the molar concentrations of oleic acid are 0.125 mmol/L and 0.25 mmol, respectively. /L, 0.5mmol/L, 1.0mmol/L, 2.0mmol/L.

Viability assay: Take the HepG2 cells treated with Chinese medicine composition 1, Chinese medicine composition 2 or oleic acid, remove the cell supernatant, and inject 100 μL of 0.05 g/mL thiazole blue solution into each well, and continue at 37°C After incubation for 30 min, HepG2 cells were washed twice with PBS, and 100 μL of dimethyl sulfoxide was added to each well to dissolve the formazan in the cells. After shaking for 20 min, the optical density of the dimethyl sulfoxide solution was measured at a wavelength of 50 nm.

Taking no traditional Chinese medicine composition and oleic acid as the control group, taking dimethyl sulfoxide as the solvent blank as the control.

Experimental results: As shown in Figure 1, in the mass concentration range of 0-500 μg/mL, the traditional Chinese medicine composition 1 and the traditional Chinese medicine composition 2 have different degrees of inhibition on the activity of pancreatic enzymes, and the traditional Chinese medicine composition 1 has the effect of inhibiting the activity of pancreatic lipase. The inhibition is strong, and with the increase of mass concentration, the inhibition effect is obviously improved.

As shown in Figure 2, after treating HepG2 cells with oleic acid-containing growth medium, the activity of HepG2 cells was significantly inhibited, and when the concentration of oleic acid was too high, it was more toxic to HepG2 cells. The molar concentration of oleic acid was 0.125-0.5mmol/L cultured cells is the best.

(3) Influence experiment of Chinese medicine composition 1 and Chinese medicine composition 2 prepared in Example 1 on triglyceride synthesis in high-fat HepG2 cell model

Dissolve the traditional Chinese medicine composition 1 or 2 in DMSO with a molar concentration of 10 ^-2 M, dilute to 10 ^-5 M, and inoculate HepG2 cells at 3×10 ⁵ cells/well into a 96-well plate. After washing HepG2 cells in DMEM medium without Chinese medicine composition, they were treated with Chinese medicine composition 1 or Chinese medicine composition containing different mass concentrations (0, 10 μg/mL, 50 μg/mL, 100 μg/mL, 200 μg/mL, 500 μg/mL) 2 in DMEM medium, incubated at 37°C for 12h, then removed the cell supernatant, and continued to culture with oleic acid-containing induction medium for 24h, and normal HepG2 cells cultured in DMEM medium for 36h were the blank group.

After the incubation, the cells were gently washed with PBS for 3 times, then the cells were lysed with RIPA lysis solution in an ice bath for 30 min, and then centrifuged at 4 °C and 10000 rpm for 15 min. , and the triglyceride content in the supernatant was determined using a triglyceride detection kit, and the result was calculated as the amount of protein.

Experimental results: As shown in Figure 3, compared with the control results of the blank group, the content of triglycerides in the high-fat HepG2 cell model induced by oleic acid was reduced after treatment with Chinese medicine composition 1 and Chinese medicine composition 2, which indicated that Both Chinese medicine composition 1 and Chinese medicine composition 2 have inhibitory effects on triglyceride synthesis in high-fat HepG2 cells, and Chinese medicine composition 1 has the strongest inhibitory activity on triglyceride synthesis in high-fat HepG2 cells.

(4) Influence experiment of traditional Chinese medicine composition 1 prepared in Example 1 on lipid synthesis in high cholesterol HepG2 cell model

The traditional Chinese medicine composition 1 is dissolved in DMSO with a molar concentration of 10 ^-2 M, and diluted to 10 ^-5 M. The DMSO is used as a solvent, which is helpful for the penetration of the traditional Chinese medicine composition 1, and is also an osmotic protective agent. , take HepG2 cells at 3×10 ⁵ cells/well and inoculate them into a 96-well plate, wait for them to adhere, remove the cell supernatant, and use cells containing different mass concentrations (0, 10 μg/mL, 50 μg/mL, 100 μg/mL, 200 μg/mL, 500 μg/mL) traditional Chinese medicine composition 1 in DMEM culture was incubated at 37°C for 12 hours, the cell supernatant was removed, and then cultured with cholesterol-containing induction medium for 24 hours, with DMEM medium for 12 hours, and the cell supernatant was removed. The cells were further cultured for 24 h with the medium containing 15 mg/mL cholesterol and 0.2 mg/mL simvastatin as the simvastatin positive control group.

After the incubation, the cells were gently washed with PBS for 3 times, and the cells were lysed with RIPA lysis solution in an ice bath for 30 min, centrifuged at 4°C and 10,000 rpm for 15 min, and the supernatant was collected. The content of total cholesterol in HepG2 cells was measured with a total cholesterol detection kit, and the results were calculated as protein quality.

Experimental results: According to the above experimental results, the traditional Chinese medicine composition 1 with the strongest inhibitory effect on triglyceride in the high-fat HepG2 cell model was selected, and the effect of lipid synthesis in the high-cholesterol HepG2 cell model was carried out. Compared with the control, both 0.2mg/mL simvastatin and Chinese medicine composition 1 can significantly reduce the content of total cholesterol in the HepG2 cell model, which indicates that simvastatin and Chinese medicine composition 1 can effectively inhibit the high cholesterol HepG2 cell model. Lipid synthesis, and with the increase of the concentration of traditional Chinese medicine composition 1, its inhibitory activity on lipid synthesis in the high cholesterol HepG2 cell model also gradually increased, and the inhibitory effect was the best when its mass concentration was 500 μg/mL.

(5) Influence experiment of the traditional Chinese medicine composition prepared in Example 2-7 on the viability of HepG2 cells

After HepG2 cells were trypsinized, 3×10 ⁵ cells/well were seeded into 96-well plates, incubated at 37°C for 24 h, and the cell supernatant was removed after the incubation. The growth medium of the traditional Chinese medicine composition was incubated at 37°C for 24 hours. The traditional Chinese medicine composition prepared in each example was set up in 5 experimental groups, and the mass concentrations of the traditional Chinese medicine composition were 0, 10 μg/mL, 50 μg/mL, and 100 μg respectively. /mL, 200μg/mL, 500μg/mL;

Viability assay: Take the HepG2 cells treated with the traditional Chinese medicine composition, remove the cell supernatant, inject 100 μL of 0.05 g/mL thiazole blue solution into each well, continue to incubate at 37 °C for 30 min, and then wash the HepG2 cells with PBS Twice, 100 μL of dimethyl sulfoxide was added to each well to dissolve the formazan in the cells. After shaking for 20 min, the optical density of the dimethyl sulfoxide solution was measured at a wavelength of 50 nm to obtain the effect of traditional Chinese medicine compositions on pancreatic fat in each experimental group. The inhibition data of enzymatic activity are shown in Table 1:

Table 1 Inhibitory data of traditional Chinese medicine composition on pancreatic lipase activity

Example 10

Weigh 1 100 g of traditional Chinese medicine composition, 50 g of dextrin, 10 g of sodium carboxymethyl cellulose, and 2 g of magnesium stearate, mix the substances, pass through an 80-mesh sieve, use a dry granulator to granulate, and sieve with a 60-mesh sieve. After granulation, the capsules are filled into capsules to obtain traditional Chinese medicine composition capsules, wherein dextrin is used as excipient and filler, sodium carboxymethyl cellulose is used as thickening agent, and magnesium stearate is used as glidant.

Example 11

Weigh 1 100 g of traditional Chinese medicine composition, 30 g of dextrin, 10 g of sodium carboxymethyl cellulose, 20 g of sodium hydroxypropyl cellulose, and 2 g of magnesium stearate, mix the substances, pass through an 80-mesh sieve, and use dry granulation Machine-made granulation, granulating with a 60-mesh sieve, tableting, and coating to obtain traditional Chinese medicine composition tablets, wherein dextrin is used as excipient and filler, sodium carboxymethyl cellulose is used as thickening agent, and hydroxypropyl fiber Sodium is used as the film coating material and binder of the tablet, and magnesium stearate is used as a glidant in the tableting process. If the dosage of the above-mentioned additives is too small, the tablet molding will be unstable during the tableting process and affect the quality of the tablet. , too much will affect the content of serum extract in the tablet and reduce the efficacy of the tablet.

The 100 volunteers were divided into three groups, the first 40 volunteers took the traditional Chinese medicine composition tablets prepared in Example 11, three times a day, 2-4 tablets each time, and the 41st to 80th volunteers took The capsules of the traditional Chinese medicine composition prepared in Example 10, three times a day, 2-4 capsules each time, the 81st to the 100th volunteers took the traditional Chinese medicine composition 1 prepared in Example 1 100g, three times a day, each time 2- 4 copies, continuous medication for observation for one year, the experimental grouping and efficacy parameters are shown in Table 2, of which 88 cases were markedly effective and 12 cases were effective. All volunteers' symptoms of hypertension and hyperlipidemia were well relieved. The blood lipid data of the patients before and after are shown in Table 3. No obvious toxic and side effects were found in the application patients, and very few people with cold stomach had mild diarrhea, which could be corrected by eating jujube and other warm medicines and foods at the same time. The traditional Chinese medicine composition 1 and the prepared tablets and capsules can effectively reduce the blood pressure and blood lipids of patients, and the action process is mild and the side effects are small.

Table 2 Experimental grouping and validity parameters

group Number of cases (person) Effective (person) Effective (person) total efficiency 1 40 36 4 100% 2 40 32 8 100% 3 20 18 2 100%

Table 3 Blood pressure and blood lipid values of volunteers before and after the experiment

See serum is a cold Chinese herbal medicine with complex chemical composition. It is directly used for lowering blood pressure and blood lipid without separation and purification, which will cause irritation to the gastrointestinal tract, especially for people with weak gastrointestinal function. Symptoms such as vomiting may also lead to decreased appetite, diarrhea, etc., and the effect of lowering blood pressure and blood lipid is not significant, and the effect is slow. The anthraquinones, including chrysophanol, emodin methyl ether, cassia, orange cassia, etc., supplemented by the full-component extracts of hawthorn and wolfberry, are compounded to obtain a traditional Chinese medicine composition. Cassia has a laxative effect, and hawthorn extracts The extract is rich in fruit acids and organic acids, which can stimulate the secretion of digestive juice, improve the function of the human spleen and stomach, and relieve symptoms such as diarrhea and loss of appetite. In addition to the effects of nourishing yin, invigorating the kidney, and soothing the nerves, the wolfberry extract also contains wolfberry polysaccharide, which has a two-way The effect of regulating blood sugar; To sum up, the effects of the various substances in the traditional Chinese medicine composition provided in this embodiment can complement each other, realize the effect of lowering blood lipid and blood pressure in a short time, and avoid the side effects caused by ineffective ingredients, stimulating ingredients, etc. , verified by volunteers, the blood pressure and blood lipid lowering effect of the traditional Chinese medicine composition described in this example is extremely significant, and it can restore normal blood pressure in volunteers within 2 weeks, and does not rebound within 2 months of drug withdrawal.

Each embodiment in this specification is described in a related manner, and the same and similar parts between the various embodiments may be referred to each other, and each embodiment focuses on the differences from other embodiments. In particular, for the system embodiments, since they are basically similar to the method embodiments, the description is relatively simple, and for related parts, please refer to the partial descriptions of the method embodiments.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the protection scope of the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention are included in the protection scope of the present invention.

Claims (9)

1. The preparation method of the traditional Chinese medicine composition is characterized by comprising the following steps:

step 1, cleaning dried serum, removing impurities, sieving, repeatedly extracting with an acidic polar solution, combining extracting solutions, neutralizing with a dilute alkali solution to make the pH value of the system be 6.5-7.5, and recovering a polar solvent in vacuum to obtain an extract;

step 2, dispersing the extract with water, adding ethanol for precipitation for 24 hours, filtering, recovering ethanol from filtrate, extracting the ethanol extract with chloroform, and separating and purifying the extracted components to obtain a serum extract;

and 3, respectively extracting the cassia seed extract, the hawthorn extract and the medlar extract, and mixing the serum extract and the extracts to obtain the traditional Chinese medicine composition.

2. The method for preparing a Chinese medicinal composition according to claim 1, wherein the specific process of repeatedly extracting in step 1 is as follows:

extracting for 1-10 h at 25-90 ℃ by using an acidic polar solvent, and extracting for 0.5-1.5 h at 25-90 ℃ by using an acidic polar solvent, wherein the pH value of the acidic polar solvent is 1.0-3.0.

3. The method for preparing a Chinese medicinal composition according to claim 1, wherein the separation and purification process in step 2 is as follows:

performing chromatography on silica gel column using CHCl₃Performing gradient elution with/MeOH as elution system, and collecting active compounds by parts to obtain the product, i.e. the extract of the blood serum.

4. The method for preparing a Chinese medicinal composition according to claim 1, wherein the process for extracting the cassia seed extract comprises the following steps:

drying and pulverizing semen Cassiae, wetting with 15% sulfuric acid solution for 5d, reflux-extracting with chloroform for 3h, cooling slightly, vacuum-filtering, separating acid water layer from extractive solution, and repeatedly washing with distilled water to obtain chloroform extractive solution;

repeatedly extracting the chloroform extract with a buffer solution with a pH of 8.0 to obtain a raffinate phase, and repeatedly extracting the raffinate phase with a 0.2% NaOH solution;

mixing NaOH extraction liquid, dropwise adding hydrochloric acid under the stirring condition until the pH value of the system is 3, performing suction filtration after complete precipitation, washing the precipitate to be neutral by using distilled water, and drying;

dissolving the precipitate with acetone, performing chromatography with silica gel column, eluting with petroleum ether-ethyl acetate, collecting yellow color band, mixing the fractions, concentrating, standing for crystallization, collecting crystal, repeating column chromatography for separation, and recrystallizing with acetone to obtain semen Cassiae extract.

5. The method for preparing a Chinese medicinal composition according to claim 1, wherein the extraction process of the hawthorn extract and the medlar extract is as follows:

soaking fructus crataegi or fructus Lycii in distilled water for 30min, heating to boil, slightly boiling for 60min, press filtering, decocting fructus crataegi or fructus Lycii in distilled water for 30min, mixing decoctions, vacuum concentrating, and slowly concentrating in 50 deg.C water bath to obtain fructus crataegi extract or fructus Lycii extract.

6. The traditional Chinese medicine composition prepared by the preparation method of the traditional Chinese medicine composition according to any one of claims 1 to 5, which is characterized in that the mass ratio of the raw materials based on raw materials is as follows: see serum 1000: 100-2000 parts of cassia seed: 10-1000 parts of hawthorn: 5-500 of medlar.

7. An oral preparation made from the Chinese medicinal composition of claim 6, wherein the oral preparation is a tablet or a capsule.

8. The traditional Chinese medicine composition oral preparation according to claim 7, wherein the oral preparation is a tablet and is prepared from the following substances in parts by mass: 10 of the traditional Chinese medicine composition: dextrin 3: sodium carboxymethylcellulose 1: sodium hydroxypropyl cellulose 2: and 0.2 of magnesium stearate.

9. The traditional Chinese medicine composition oral preparation according to claim 7, wherein the oral preparation is a capsule prepared from the following substances in parts by mass: 10 of the traditional Chinese medicine composition: dextrin 5: sodium carboxymethylcellulose 1: and 0.2 of magnesium stearate.

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