Traditional Chinese medicine composition for treating kidney micro abdominal mass and damp toxin exuberance type membranous nephropathy as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention provides a traditional Chinese medicine composition for treating kidney micro abdominal mass and damp toxin exuberance type membranous nephropathy as well as a preparation method and application of the traditional Chinese medicine composition. The invention provides a traditional Chinese medicine composition for treating kidney micro abdominal mass and damp toxin accumulation type membranous nephropathy. The traditional Chinese medicine composition comprises the following raw materials: rhizoma dioscoreae nipponicae, euonymus alatus and oldenlandia diffusa in a mass ratio of 1: 0.6: 0.6. The preparation method adopts a water decoction extraction process and specifically comprises the following steps of soaking and water decoction, during water decoction, the water adding amount is 10-12 times of the amount of all the raw materials, the decoction time is 1.5-2 h, and the decoction frequency is 2-3 times. The traditional Chinese medicine composition for treating kidney microabdominal mass and damp toxin accumulation type membranous nephropathy provided by the invention has the advantages that the kidney microabdominal mass and damp toxin accumulation type membranous nephropathy can be effectively treated; the traditional Chinese medicine composition prepared by adopting the preparation method provided by the invention has higher safety, higher paste yield and higher dioscin content.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine, and in particular to a traditional Chinese medicine composition for treating renal micro-mass and dampness-toxin accumulation type membranous nephropathy, as well as a preparation method and application thereof. Background Art

[0002] At present, Western medicine has limited knowledge about membranous nephropathy. The KDIGO guidelines recommend that clinicians use immunosuppressants combined with glucocorticoids to alleviate the progression of kidney damage. Due to the particularity of immunosuppressants and glucocorticoids, while relieving symptoms, they can cause a series of adverse reactions such as liver and kidney damage. Therefore, the use of traditional Chinese medicine to treat membranous nephropathy is gradually being promoted.

[0003] At present, Chinese medicine generally believes that the pathogenesis of membranous nephropathy is that the kidney stores essence and controls qi. Kidney qi is weak and cannot be fixed, and the ability of qi transformation and transpiration decreases. Therefore, the fine substances are lost with urine, and eventually a large amount of proteinuria is formed. It is often accompanied by clinical symptoms such as edema of both lower limbs, soreness of waist and knees, tinnitus and deafness, shortness of breath and fatigue, poor appetite and bland mouth without thirst, and frequent urination at night. Therefore, most doctors currently choose to strengthen the spleen and tonify the kidney as the main method, supplemented by removing dampness and turbidity, to achieve the effect of dredging collaterals, replenishing deficiency and purging excess, effectively controlling proteinuria and delaying kidney damage. However, the Chinese medicine prescriptions obtained according to these treatment principles are not suitable for membranous nephropathy with kidney micro-complications and dampness and toxicity. Summary of the invention

[0004] The present invention aims to solve at least one of the technical problems in the prior art to a certain extent. To this end, in a first aspect of the present invention, the present invention provides a Chinese medicine composition for treating renal micro-mass and dampness-toxicity-prone membranous nephropathy, the raw materials of which include: Rhizoma Dioscoreae, Radix Psoraleae and Herba Hedyotis Diffusae in a mass ratio of 1:0.6:0.6.

[0005] The inventor has found in clinical work for many years that membranous nephropathy is often seen in the syndrome type of "kidney micro-symptoms and masses, dampness and toxins are congested". The kidney is responsible for storing essence. People with kidney deficiency are susceptible to external evils such as wind, dampness, heat, toxins, and blood stasis, or endogenous evil qi. The evil qi is solid, stays for a long time, and stays in the kidney, forming "kidney excess syndrome". Patients with membranous nephropathy often suffer from exogenous wind-heat evil, which causes heat evil to enter the body to transform toxins. The nature of the poison is fiery and congested, which can easily burn the kidney network, or exogenous damp-heat evil, water and dampness stagnate, block the qi mechanism, dampness turns into heat, consumes qi and hurts body fluid, causes damage to the veins and collaterals, forces blood to flow recklessly, and blood overflows outside the veins. If heat enters the blood, the qi mechanism stagnates, there is no water to travel, and blood stasis is blocked in the kidney network, and even blocked. The clinical manifestations are proteinuria and edema, and they are often repeated due to exogenous factors, and are difficult to heal. Combined with modern pathological morphology, it is believed that the pathological manifestations of dampness, toxicity and blood stasis are "micro-masses", namely the so-called glomerular sclerosis, mesangial cell hyperplasia, platelet aggregation, abnormal coagulation function, hyperlipidemia, etc. Once the micro-masses of the kidneys are produced, they are difficult to eliminate. "Dampness", "stasis" and "toxicity" are both the cause and the result of this disease, affecting each other throughout the whole process. The treatment method should be to activate blood circulation, eliminate symptoms, and promote dampness and detoxification.

[0006] Rhizoma Dioscoreae belongs to the Dioscoreaceae family. It is slightly cold in nature and bitter in taste. It enters the liver, kidney and lung meridians and has the effects of dispelling wind and dampness, activating blood circulation and unblocking collaterals. Rhizoma Dioscoreae “can defend and move, can replenish and unblock”, that is, under certain dosage and compatibility conditions, it can not only dispel wind and dampness, activate blood circulation and unblock collaterals, but also support the body’s resistance; it has the effects of reducing urine protein levels, eliminating edema, preventing thrombosis, and improving renal blood flow. Modern pharmacology shows that the saponin chemical components in Rhizoma Dioscoreae play a major therapeutic role, with anti-tumor, liver protection, immune regulation, anti-inflammatory and other effects; it can also improve insulin tolerance, reduce liver damage, anti-fibrosis, anti-virus, lower uric acid, inhibit atherosclerosis (AS) and other wide-ranging pharmacological activities.

[0007] The function of the herb Guijianyu is to break blood, dredge menstruation, detoxify and reduce swelling. The "Yao Xing Lun" records: "Guijianyu can break old blood, can cause miscarriage, and is the main cause of waist and abdominal pain and hundreds of evil spirits." Pharmacological studies have shown that the effective ingredients extracted from the Chinese herbal medicine Guijianyu include total flavonoids, total saponins, polysaccharides and other components, which have been confirmed in the research of diabetes and cardiovascular and cerebrovascular fields. The complex components of its water extract protect the liver through various pathways such as anti-oxidation, anti-inflammatory, and anti-fibrosis. It has been verified that the degree of renal fibrosis in rats with glomerular sclerosis was reduced after treatment with Guijianyu, the protein level of rats was reduced, and the blood lipid level was improved. The analysis of its mechanism of action is that Guijianyu can inhibit the abnormal expression of fibrosis-related proteins, reduce hyperplasia, and improve glomerular sclerosis.

[0008] The shape of Hedyotis diffusa is like a snake spitting out its tongue, with strong bias, bitter and sweet taste, and cold nature. It can clear away heat and detoxify, eliminate carbuncle and disperse knots, etc. The use of Hedyotis diffusa in large doses has a miraculous effect of attacking poison. Clinical and pharmacological studies have shown that Hedyotis diffusa contains a variety of components. It is currently found that the flavonoid components of Hedyotis diffusa not only have a high proportion and are easy to purify, but also have strong drug activity. It plays an important role in the prevention and treatment of diseases such as inflammation and tumors. The mechanism involves multiple signal pathways such as NF-κB, MAPK, STAT3, TGF-β / Smad, etc. The total flavonoids of Hedyotis diffusa can improve the proportion of T lymphocyte subsets, regulate the expression of inflammatory cytokines, and enhance the body's immune function; the polysaccharide and flavonoid components of Hedyotis diffusa can improve the immune system function indicators of mice, and a comparison shows that the polysaccharide components have a more obvious effect on regulating the immune system than flavonoids.

[0009] In summary, combined with basic research such as modern pharmacology, it can be considered that in the Chinese medicine composition provided by the present application, Rhizoma Dioscoreae is the main medicine, which has the effects of dispelling wind and dampness, activating blood circulation and unblocking collaterals, and has the functions of attacking, supplementing and promoting circulation; Herba Hedyotis Diffusae is the auxiliary medicine, and its miraculous effect of attacking poison can not only remove the poison stuck in dampness, but also clear the latent heat in the blood; Herba Epimedii has a strong blood-breaking power, which can eliminate all kinds of evil and break up tumors. When used together with the first two, the effect is more severe, which can remove dampness and toxins, clear blood stasis and heat, and unblock the kidney collaterals, thereby improving the body's immunity, improving renal microcirculation, inhibiting inflammatory response, controlling protein leakage, repairing the kidneys and other multiple effects.

[0010] The Chinese medicine composition provided by the present invention can promote blood circulation, eliminate symptoms, promote dampness and detoxify, and is particularly suitable for kidney micro-symptoms and masses, and dampness and toxin congestion syndrome. It can effectively eliminate micro-symptoms and masses, and dampness and toxin congestion, and improve coagulation function, hyperlipidemia, platelet aggregation, etc.

[0011] In some embodiments, the Chinese medicine composition comprises, by weight, 300-500 parts of Herba Hedyotis diffusae, 300-500 parts of Herba Hedyotis diffusae and 550-650 parts of Herba Dioscoreae.

[0012] In some embodiments, the dosage form of the Chinese medicine composition is selected from at least one of decoction, powder, granule, and capsule, preferably capsule.

[0013] Based on this, the present invention also provides a preparation method of the Chinese medicine composition, which adopts a water decoction extraction process, specifically comprising the following steps: soaking and water decoction, wherein the soaking time is 10-30 minutes, when the water is decocted, the amount of water added is 10-12 times the amount of all raw materials, the decoction time is 1.5-2 hours, and the decoction times are 2-3 times. The Chinese medicine composition prepared by the preparation method has higher safety, higher cream yield and higher dioscin content.

[0014] In some embodiments, the method further comprises concentration, drying and pulverization, wherein the concentration is performed at 60-70° C. to a relative density of 1.23-1.28, the drying temperature is 60-65° C., and after the pulverization, a fine powder of 80-100 mesh is obtained.

[0015] In some embodiments, the fine powder is further filled into capsules, and the content of the traditional Chinese medicine composition in each capsule is 0.4-0.6g.

[0016] In some embodiments, the fine powder is further filled into capsules, and the content of the traditional Chinese medicine composition in each capsule is 0.5 g.

[0017] In a third aspect, the invention provides the use of the Chinese medicine composition and / or the Chinese medicine composition obtained by the preparation method in drugs for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type.

[0018] In some embodiments, the Chinese medicine composition is administered orally.

[0019] In some embodiments, the dosage of the Chinese medicine composition is 0.075-0.125 g / kg / d.

[0020] The Chinese medicine composition for treating membranous nephropathy of the type with renal micro-complications and dampness and toxin accumulation provided by the present invention can reduce renal tissue pathological morphologies such as glomerular podocyte enlargement, basement membrane thickening, podocyte foot process fusion, etc., can effectively improve coagulation function, can effectively eliminate renal micro-complications and dampness and toxin accumulation, can reduce the 24hUTP quantification and mALB level of membranous nephropathy rats; reduce the levels of Cr, BUN, TC, TG, and LDL-C, and can increase the levels of ALB and TP, indicating that the Chinese medicine composition provided by the present invention can effectively treat membranous nephropathy of the type with renal micro-complications and dampness and toxin accumulation; the Chinese medicine composition prepared by the method provided by the present invention has higher safety, higher cream yield, and higher dioscin content. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0022] Figure 1 It is one of the bar graphs of the ALB results of the rats in each group involved in the Chinese medicine composition for treating renal micro-mass and damp-toxin stagnation type membranous nephropathy provided by the present invention.

[0023] Figure 2 It is one of the bar graphs of TP results of each group of rats involved in the Chinese medicine composition for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type provided by the present invention.

[0024] Figure 3 It is one of the histograms of AST results of each group of rats involved in the traditional Chinese medicine composition for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type provided by the present invention.

[0025] Figure 4 It is one of the bar graphs of ALT results of each group of rats involved in the traditional Chinese medicine composition for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type provided by the present invention.

[0026] Figure 5 It is one of the bar graphs of TC results of rats in each group involved in the Chinese medicine composition for treating renal micro-mass and damp-toxin stagnation type membranous nephropathy provided by the present invention.

[0027] Figure 6This is one of the bar graphs of TG results of each group of rats involved in the traditional Chinese medicine composition for treating renal micro-mass and damp-toxin accumulation type membranous nephropathy provided by the present invention.

[0028] Figure 7 It is one of the LDL-C result bar graphs of each group of rats involved in the traditional Chinese medicine composition for treating renal micro-mass and damp-toxin stagnation type membranous nephropathy provided by the present invention.

[0029] Figure 8 It is one of the Cr result bar graphs of each group of rats involved in the traditional Chinese medicine composition for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type provided by the present invention.

[0030] Fig. 9 It is one of the bar graphs of the BUN results of the rats in each group involved in the Chinese medicine composition for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type provided by the present invention.

[0031] Fig.10 This is one of the result graphs (HE staining, ×200) showing the effect of the Chinese medicine composition for treating membranous nephropathy of renal micro-mass and dampness-toxin accumulation type provided by the present invention on the renal tissue pathology of rats with membranous nephropathy.

[0032] Fig.11 This is one of the results of the effect of the Chinese medicine composition for treating membranous nephropathy of renal micro-mass and dampness and toxin accumulation type provided by the present invention on the renal tissue pathology of rats with membranous nephropathy (Masson staining, ×200).

[0033] Fig.12 This is one of the results of the effect of the Chinese medicine composition for treating membranous nephropathy of renal micro-mass and dampness and toxin accumulation type provided by the present invention on the renal tissue pathology of rats with membranous nephropathy (PASM staining, ×200).

[0034] Fig.13 This is one of the scanning electron microscope images (×10000) of the effect of the Chinese medicine composition for treating membranous nephropathy of renal micro-mass and dampness and toxin accumulation type provided by the present invention on the ultrastructure of renal tissue of rats with membranous nephropathy.

[0035] Fig.14 This is one of the transmission electron microscopic images (×3000) of the effect of the Chinese medicine composition for treating membranous nephropathy of renal micro-mass and dampness and toxin accumulation type provided by the present invention on the ultrastructure of renal tissue of rats with membranous nephropathy. DETAILED DESCRIPTION

[0036] Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making any creative work shall fall within the scope of protection of the present invention.

[0037] In the first aspect of the present invention, a Chinese medicine composition for treating renal micro-mass and damp-toxic congestion type membranous nephropathy is proposed, wherein the raw materials include: Rhizoma Dioscoreae, Radix Glehniae and Herba Hedyotis Diffusae in a mass ratio of 1:0.6:0.6; in some embodiments, the raw materials include: 360 parts of Radix Glehniae, 360 parts of Herba Hedyotis Diffusae and 600 parts of Rhizoma Dioscoreae by weight. In some embodiments, 1 part is equal to 1g, such as 360g of Radix Glehniae, 360g of Herba Hedyotis Diffusae and 600g of Rhizoma Dioscoreae.

[0038] In the formula, Rhizoma Cynoglossi is the main ingredient, which has the effects of dispelling wind and dampness, activating blood circulation and unblocking meridians, and has the functions of attacking, supplementing, and promoting circulation. Herba Hedyotis diffusae is the auxiliary ingredient, which has the miraculous effect of attacking toxins, which can not only remove the toxins stuck in dampness, but also clear the latent heat in the blood. Herba Epimedii has a strong blood-breaking power, and is said to be able to eliminate all kinds of evils and break up tumors. When used together with the first two, the effect is more powerful, which can remove dampness and toxins, clear away blood stasis and heat, and unblock the kidney meridians, thereby enhancing the body's immunity, improving renal microcirculation, inhibiting inflammatory response, controlling protein leakage, and repairing the kidneys.

[0039] Chinese patent application CN103041099A discloses a drug for treating chronic kidney disease with blood heat and damp-heat toxicity, which is made of total flavonoids of Psoralea corylifolia, polysaccharides of Hedyotis diffusa, total saponins of Rhizoma Dioscoreae and pharmaceutical excipients. Although there is no clear explanation of the prescription, it can be seen from the fact that the syndromes treated are blood heat and damp-heat toxicity, and the main drug is Hedyotis diffusa with the effect of clearing away heat, promoting dampness and detoxifying. In the present invention, the syndromes treated are mainly membranous nephropathy with kidney micro-mass and damp-toxicity congestion, and the Chinese medicine composition uses Rhizoma Dioscoreae with the effect of promoting blood circulation as the main drug, which is completely different from the drugs in the above-mentioned prior art.

[0040] In some embodiments, the dosage form of the Chinese medicine composition is selected from at least one of decoction, powder, and capsule, preferably capsule, which has the advantages of masking the odor of the medicine, less irritation to the stomach and intestines, and is easy to take, carry, and store.

[0041] The present invention also provides a method for preparing the Chinese medicine composition, which adopts a water decoction extraction process, wherein the amount of water added during the water decoction is 10-12 times the amount of all raw materials, the decoction time is 1.5-2h, and the decoction times are 2-3 times. The use of this water decoction process is conducive to obtaining a higher paste yield, thereby ensuring the efficacy and saving costs.

[0042] In some embodiments, the preparation method of the traditional Chinese medicine composition also includes concentration, drying and crushing, wherein the concentration is concentrated to a clear paste with a relative density of 1.23-1.28 (60°C thermal measurement) at 60-70°C (such as 60°C, 65°C, 70°C or other temperatures, or a range composed of these temperatures), the drying temperature is 60-65°C (such as 60°C, 62°C, 65°C or other temperatures, or a range composed of these temperatures), and after the crushing, a fine powder of 80-100 mesh (such as 80 mesh, 90 mesh, 100 mesh or other mesh sizes, or a range composed of these mesh sizes) is obtained.

[0043] In some embodiments, the preparation method of the Chinese medicine composition further includes further filling the fine powder into capsules, and the content of the Chinese medicine composition per capsule is 0.4-0.6g, such as 0.4g / capsule, 0.5g / capsule, 0.6g / capsule or other contents, or a range of these contents.

[0044] In a third aspect, the invention provides the use of the Chinese medicine composition and / or the Chinese medicine composition obtained by the preparation method in drugs for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type.

[0045] In some embodiments, the Chinese medicine composition is administered orally. For example, it is made into an oral dosage form such as a decoction or a powder, and flavor and odor regulating ingredients can be appropriately added according to needs.

[0046] In some embodiments, the dosage of the Chinese medicine composition is 0.075-0.125 g / kg / d, such as 0.075 g / kg / d, 0.08 g / kg / d, 0.09 g / kg / d, 0.1 g / kg / d, 0.11 g / kg / d, 0.12 g / kg / d, 0.125 g / kg / d or other dosages, or a range consisting of these dosages.

[0047] In the embodiment, the instrument information is shown in Table 1 below.

[0048] Table 1 Experimental instruments In the examples, the experimental drugs and actual information are shown in Table 2 below.

[0049] Table 2 Experimental reagents Chinese herbal medicines Rhizoma Cynoglossi (20191101), Psoralea corylifolia (20190901), and Hedyotis diffusa (20190801) were purchased from the pharmacy of Shanxi Provincial Hospital of Traditional Chinese Medicine and identified as authentic by Professor Liu Guangzhen of Shanxi Provincial Hospital of Traditional Chinese Medicine.

[0050] Tripterygium wilfordii glycosides tablets were purchased from Yuanda Pharmaceutical Huangshi Feiyun Pharmaceutical Co., Ltd., item number: Z42021212.

[0051] Preparation of cationic bovine serum albumin: According to the modified Border method, 67 mL of anhydrous ethylenediamine solution (EDA) was added to 500 mL of double distilled water and mixed; 6 mol / L hydrochloric acid was prepared into 300 mL, and the pH value was adjusted to 4.75 by a pH tester, cooled to 25 °C, and stirred with a glass rod; 5 g BSA was weighed on a balance and fully dissolved in 25 mL of double distilled water; then carbodiimide (EDC) was added and stirred thoroughly, and after stirring thoroughly with a magnetic stirrer for 2 h, the pH value was adjusted to 4.75; then 30 ml of acetate buffer was slowly added to terminate the reaction, and the pH value of the solution was finally adjusted to 4.75 to complete the preparation of C-BSA; finally, the prepared C-BSA solution was slowly poured into a dialysis bag, and dialyzed continuously with double distilled water for 72 h, and after vacuum freeze drying, it was dried into C-BSA dry powder and placed in a -20 °C refrigerator for use.

[0052] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0053] The present invention is described below with reference to specific embodiments. It should be noted that these embodiments are merely illustrative and do not limit the present invention in any way.

[0054] Example 1 The present embodiment provides a traditional Chinese medicine composition for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type, which is composed of three traditional Chinese medicines: Rhizoma Dioscoreae, Radix Psoraleae and Herba Hedyotis Diffusae in a ratio of 1:0.6:0.6, specifically: 600g of Rhizoma Dioscoreae, 360g of Radix Psoraleae and 360g of Herba Hedyotis Diffusae.

[0055] This embodiment also provides a method for preparing the Chinese medicine composition, comprising the following steps: (1) Extraction: Soak the three herbs in the prescription for 15 minutes, add 12 times the amount of water, and boil twice, each time for 2 hours, to obtain the water extract; (2) Concentration: The water extracts are combined and filtered, and heated and concentrated to a clear paste with a relative density of 1.23 to 1.28 (thermal measurement at 60°C), and the concentration temperature is controlled at about 65°C; (3) Drying: evenly spread the clear paste in a stainless steel plate and dry it at 60-65°C to obtain a dry paste; (4) Grinding: Grinding the dry paste into 80 mesh fine powder; (5) Filling: Add the fine powder into a fully automatic capsule filling machine for filling, with a specification of 0.5 g / capsule.

[0056] Test Example 1 Animal Experiment 1. Experimental subjects: 66 specific pathogen-free (SPF) male SD (Sprague Dawley) rats, about 6 weeks old, weighing 160-180 g (Beijing Huafukang Biotechnology Co., Ltd., license number: SCXK Jing 2017-0005), temperature 20-25 ° C, relative humidity 35%-60%, fed with ordinary feed (Beijing Huafukang Biotechnology Co., Ltd.), free access to food and water, and the experiment was conducted after 1 week of routine adaptive feeding in the same standard environment.

[0057] 2. Test methods: 2.1 Establishment of rat membranous nephropathy model Ten rats were randomly selected as the blank group, and all other rats were modeled.

[0058] First, pre-immunization: cationic bovine serum albumin was injected into the tail vein to induce a rat model of membranous nephropathy. 1 mg of cationic bovine serum albumin dry powder was thoroughly mixed with 0.5 mL of phosphate buffer, and then a 1:1 volume of Freund's incomplete adjuvant was added and stirred thoroughly to emulsify. 0.5 mL of the emulsion was injected subcutaneously into the left and right armpits, left and right groin, and back of the rat, once every 2 days, for a total of 3 times.

[0059] Secondly, the formal immunization was carried out. The C-BSA dry powder was completely dissolved in phosphate buffered saline and injected into the tail vein. There was no stagnant resistance to the needle during the injection, indicating that the injection was successful. Once every two days for a total of four weeks, the urine protein determination kit was used to conduct a qualitative test of rat urine protein. The standard for successful model preparation was that the rat 24hUTP ≥ 20mg.

[0060] 2.2 Grouping and intervention Grouping: After removing the dead rats and the rats with unsuccessful modeling, the prepared rats were randomly divided into a model group of 9 rats, a Tripterygium wilfordii polyglycosides tablets positive group of 9 rats, and a high-dose, low-dose, and medium-dose test sample group of 9 rats each.

[0061] Grouping and drug administration: After the test model is successfully established, oral administration begins (the dosage is converted into an equivalent dosage based on the body surface area of ​​humans and rats, and combined with the results of previous experiments and pre-experiments as follows) (1) Blank group: The same volume of normal saline was administered as in the treatment group, once a day, for 8 weeks; (2) Model group: The same volume of normal saline was administered as in the treatment group, once a day, for 8 weeks; (3) High-dose group: 1.5 g / kg / d solution, once a day, for 8 weeks; (4) Medium-dose group: 0.75 g / kg / d solution, once a day, for 8 weeks; (5) Low-dose group: 0.375 g / kg / d solution, once a day, for 8 weeks; (6) Tripterygium wilfordii glycosides control group: 7.5 mg / kg / d solution was administered once a day for 8 weeks.

[0062] In the above, the test sample is the Chinese medicine composition provided in Example 1.

[0063] 2.3 Result detection 2.3.1 General condition of rats with membranous nephropathy Observe and record the rats' daily mental state, hair, reaction sensitivity, food and water intake, feces and bowels, body weight and other general conditions.

[0064] 2.3.2 Detection of urine protein in rats with membranous nephropathy Urine was collected from rats at the end of the 0th, 4th, and 8th week. Urine was collected in 15 ml centrifuge tubes and the urine volume was recorded. 24hUTP was measured using a urine protein detection kit and mALB was detected using ELISA. The operation was performed strictly according to the instructions of the kit.

[0065] Enzyme-linked immunosorbent double antibody sandwich method (ELISA) detection of mALB: dilute the standard with gradient concentration values, take 5 2mL EP tubes, number them from 1 to 5, EP tube ① contains 150μL of the original concentration of the standard, add 150μL of the standard diluent, and mix thoroughly with a pipette. EP tube ② contains 150μL of the standard ①, add 150μL of the standard diluent, and mix thoroughly with a pipette. EP tube ③ contains 150μL of the standard ②, add 150μL of the standard diluent, and mix thoroughly with a pipette. EP tube ④ contains 150μL of the standard ③, add 150μL of the standard diluent, and mix thoroughly with a pipette. EP tube ⑤ contains 150μL of the standard ④, add 150μL of the standard, add 150μL of the standard diluent, and mix thoroughly with a pipette. Dilute to the required gradient concentration value; add samples, take the enzyme-labeled coated plate, tear off the coating, add 50μL of the standard sample, and add 2 parallel wells for each concentration; add 40μL of the standard diluent to the samples to be tested, and then add 10μL of mALB sample; shake gently to mix, seal the plate with a sealing film, and incubate in a 37℃ oven for 30min; dilute the concentrated washing solution 30 times with distilled water for standby use; steadily and slowly lift the sealing film, discard and shake off the liquid, then fill the ELISA plate with a syringe to fill the washing solution, let it stand for 30s, then shake off the washing solution, repeat 5 times, and pat the liquid dry completely; add 50μL of enzyme-labeled reagent to each well except the control well; seal the plate with a sealing template and incubate in a 37℃ oven for 30min; gently and slowly tear off the sealing film of the incubated ELISA plate, discard and shake off the liquid, fill the ELISA plate with a syringe to fill the washing solution, let it stand for 30s Then shake off the washing solution vigorously and repeat 5 times; add 50μL of color developer A and 50μL of color developer B to each well in order of A and B, shake gently back and forth to mix completely, and put in a 37℃ oven for color development under light-proof conditions for 10 minutes; add stop solution to each well and stop the experiment; use the horizontal axis as the standard concentration and the vertical axis as the OD value to calculate the actual concentration of the final sample.

[0066] 2.3.3 Detection of rat blood biochemical indicators All rats were fasted for 12 hours, anesthetized and killed, and blood was collected from the abdominal aorta and allowed to stand for 30 minutes. The blood was centrifuged and separated into 2 ml EP tubes to obtain the supernatant, which was frozen in a -80°C refrigerator for testing. The biochemical indicators of creatinine (Cr), triglycerides (TG), total protein (TP), urea nitrogen (BUN), total cholesterol (TC), albumin (ALB), alanine aminotransferase (ALT), low-density lipoprotein cholesterol (LDL-C) and aspartate aminotransferase (AST) were tested using an automatic biochemical analyzer.

[0067] 2.3.4 Observation of pathological changes of rat kidney under light microscopy After blood collection, the kidneys were dissected, and partial kidney tissues of 3 rats were taken from each group, rinsed with physiological saline, and fixed in 4% paraformaldehyde fixative. The kidney tissues were removed from the fixative, and different concentrations of alcohol were added to completely dehydrate them. Then, the tissues were transparentized with xylene and kept in anhydrous state. Then, the tissues were embedded with wax to make paraffin sections (thickness of about 4 μm), and finally, paraffin sections were embedded with paraffin to make paraffin sections, which were stained with hematoxylin-eosin (HE), Masson (Masson) and iodized silver hexaamine (PASM). The sections were fully immersed in xylene for 2 h until transparent and dry, and then sealed with neutral resin. The pathological morphological changes of rat kidney tissues were observed under an optical microscope and photographed.

[0068] 2.3.5 Transmission electron microscopy to examine the ultrastructure of renal tissue Before sampling, prepare a specimen bottle filled with electron microscope fixative in advance, cut a small tissue block of 1mm×1mm×1mm from the renal cortex area of ​​the kidney tissue, clean the blood on the surface of the sample with physiological saline, absorb it with filter paper, and immediately put it in the specimen bottle (try to avoid mechanical damage to the tissue such as pulling, squeezing and contusion), and store it in a 4℃ refrigerator after 1-2 hours; then fix it for 2 hours, wash it 3 times, and fix it for 20 minutes each time; the kidney tissue is dehydrated in accordance with different concentrations of gradient alcohol and dehydrated thoroughly; then embed; stain with alcohol in the dark for 10 minutes; wash it with 70% alcohol and purified water 3 times respectively; stain it for 10 minutes, and finally wash it with purified water 3 times, and the filter paper slightly absorbs water. Dry it overnight at 37℃; finally observe it under a microscope and take pictures.

[0069] 2.3.6 Scanning electron microscopy to examine the ultrastructure of renal tissue The fixed samples were rinsed with phosphate buffer for 3 times, 15 min each time; then fixed with 1% osmic acid·0.1M phosphate buffer PB for 1-2 h at room temperature in the dark; rinsed with 0.1M phosphate buffer PB for 3 times, 15 min each time; the kidney tissue was dehydrated with different concentrations of alcohol for 15 min each time and isoamyl acetate for 15 min; the samples were placed in a critical point dryer for drying; and observed and photographed under a scanning electron microscope after conduction.

[0070] 2.4 Statistical analysis The obtained data were processed using SPSS 26.0 software, and the results were expressed as mean ± standard deviation ( ±s), all data were in accordance with normal distribution, one-way analysis of variance was used for homogeneous variance, and LSD-t test was used for multiple comparisons; rank sum test was used for comparison between groups with unequal variance, and P < 0.05 indicated statistically significant differences. GraphPad Prism9.5 software was used for graphical analysis of the obtained data.

[0071] 3 Test results: 3.1 Effect of the Chinese medicine combination on the general condition of rats with membranous nephropathy (1) The rats in the blank group were energetic, responsive, had shiny fur, moved freely, had normal diet and water intake, and their bowel movements were normal. Their body weight gradually increased.

[0072] (2) The rats in the model group were listless and slow to react. Their hair was rough and dark, and occasionally fell off. They preferred quietness and disliked movement. Their food intake was significantly reduced compared with that of the rats in the normal group. Their water intake and urine output increased. Their stools were soft and dark brown, and their body weight gradually decreased.

[0073] (3) The mental state of rats in the high, medium and low dose groups of the test substance improved compared with the modeling period. They gradually became more energetic, their reactions became more sensitive, their fur became darker in gloss, their food and water intake was normal, their urine output increased, their stools were formed and occasionally black, and their body weight gradually increased.

[0074] (4) The mental state of the rats in the control group of Tripterygium wilfordii tablets was better than that in the modeling period, their reactions became more agile, their fur became darker, their food and water intake was normal, their urine output increased, their stools were formed and black stools were less common, and their body weight gradually increased.

[0075] 3.2 Effects of Chinese herbal medicine combination on 24hUTP quantification and mALB in rats with membranous nephropathy The results of 24hUTP quantification and mALB testing are shown in Table 3.

[0076] Table 3 Test results of 24hUTP and mALB levels in rats of each group ( ) As can be seen from Table 3, after 4 weeks of drug intervention, compared with the model group, the 24hUTP of rats in the test group and the Tripterygium wilfordii polyglycosides control group began to decrease to varying degrees (P<0.05), and the decrease in rats in the test medium dose group was the most significant. After 8 weeks of drug treatment, the 24hUTP of the rat model group increased significantly compared with the normal group (P<0.01); compared with the model group, the 24h urine protein quantification of rats in the test group and the Tripterygium wilfordii polyglycosides control group decreased significantly (P<0.01).

[0077] It can be seen from Table 3 that compared with the blank group, the mALB of rats in the model group was significantly increased (P<0.01), and the difference was significant and statistically significant; after 8 weeks of drug treatment, compared with the model group, the mALB of rats in each test group was significantly decreased (P<0.01), and the mALB of rats in the Tripterygium Wilfordii Polyglycosides Tablets control group was significantly decreased (P<0.01).

[0078] Proteinuria is one of the main clinical symptoms of membranous nephropathy. It is not only a key factor in aggravating kidney disease, but also toxic to kidney disease. Therefore, reducing proteinuria is the core of treating membranous nephropathy, and it is also an important method for alleviating chronic kidney disease and the primary indicator for evaluating the therapeutic effect. The results of this experiment showed that compared with the blank group, the proteinuria level in the model group increased significantly (P<0.01); after 8 weeks of drug intervention, the 24h urine protein quantification and mALB levels of rats decreased to varying degrees, among which the 24h urine protein quantification and mALB levels were the most significant decreases in the proteinuria levels of the test sample medium dose group and the tripterygium wilfordii polyglycosides tablet control group. It fully illustrates the efficacy of the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) in reducing proteinuria in membranous nephropathy.

[0079] 3.3 Effect of Chinese herbal medicine combination on liver function in serum of rats with membranous nephropathy The test results of ALB, TP, ALT and AST are shown in Table 4. Figure 1-Figure 4 , compared with the blank group * P <0.05; ** P <0.01; compared with the model group: # P <0.05, ## P <0.01.

[0080] Table 4 Test results of ALB, TP, AST, and ALT in rats with membranous nephropathy (n=9, ) From Table 4 and Figure 1-4It can be seen that compared with the blank group, the expressions of ALT and AST in the model group were significantly increased (P<0.01), and the expressions of ALB and TP were significantly decreased (P<0.01); after 8 weeks of drug intervention, compared with the model group, the levels of TP and ALB in the rats in the high-dose group of the test sample capsule were significantly increased (P<0.01), and the level of AST was significantly decreased (P<0.05); the level of ALB in the rats in the low-dose group of the test sample was significantly increased (P<0.01), and the level of ALT was significantly decreased (P<0.05), and the level of ALB in the medium-dose group of the test sample was significantly increased (P<0.01); the levels of TP and ALB in the rats in the control group of Tripterygium wilfordii polyglycosides were significantly increased (P<0.01).

[0081] Hypoproteinemia is one of the important clinical symptoms of membranous nephropathy. During the onset of membranous nephropathy, the function of the glomerular filtration membrane is damaged, resulting in the destruction of the integrity of the filtration barrier, a large amount of protein continuously leaking from the urine, causing the loss of protein in the plasma, and a decrease in plasma colloid osmotic pressure, which can eventually lead to edema. The experimental results show that after 8 weeks of drug intervention, the ALB and TP levels of the Chinese medicine group and the control group increased to varying degrees compared with the model group (P<0.01). This shows that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) can significantly increase the serum ALB and TP levels in rats with membranous nephropathy, maintain plasma colloid osmotic pressure, improve the clinical symptoms of edema, and improve the quality of life.

[0082] ALT and AST in serum are one of the diagnostic indicators of acute viral hepatitis and are also important indicators for monitoring the disease and prognosis. Among them, ALT in serum can more sensitively reflect liver cell damage. The differences in AST and ALT levels between the test group and the control group and the model group are not obvious, indicating that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) has the advantages of high safety and no damage to liver function.

[0083] 3.4 Effect of Chinese herbal medicine combination on renal function and blood lipids in serum of rats with membranous nephropathy The test results of Cr, BUN, TC, TG, and LDL-C are shown in Table 5 and Figure 5-Figure 9 , compared with the blank group * P < 0.05; * *P<0.01; compared with the model group: # P<0.05, ## P<0.01.

[0084] Table 5 Test results of BUN, TC, Cr, TG, and LDL-C in rats with membranous nephropathy (n=9, ) From Table 5 and Figure 5-9 It can be seen that compared with the blank group, the expression of Cr, BUN, TC, TG, and LDL-C in the model group was significantly increased (P < 0.01); after 8 weeks of drug intervention, compared with the model group, the BUN in the medium-dose group of the test sample decreased significantly (P < 0.01), and Cr and TC decreased significantly (P < 0.05); the BUN, TC, and LDL-C in the high-dose group of the test sample decreased significantly (P < 0.01), and TC decreased significantly (P < 0.05); the Cr, LDL-C, and BUN in the low-dose group of the test sample decreased significantly (P < 0.05); the BUN, TC, and TG in the control group of Tripterygium wilfordii polyglycosides tablets decreased significantly (P < 0.01).

[0085] Hyperlipidemia is also one of the important clinical symptoms of membranous nephropathy. In the course of membranous nephropathy, due to the loss of proteinuria and protein, the liver compensatory effect occurs, stimulating the liver to synthesize a large amount of triglycerides, total cholesterol and lipoproteins to be transported to the blood, ultimately leading to hyperlipidemia. Dyslipidemia can accelerate glomerular mesangial proliferation, inflammatory cell infiltration and mesangial extracellular matrix deposition, etc., and ultimately increase the burden of renal function. The experimental results show that after 8 weeks of drug intervention, the TG, TC, and LDL-C levels of the test group and the control group decreased to varying degrees compared with the model group. It can be seen that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) can improve coagulation function and prevent the occurrence of complications of membranous nephropathy by regulating abnormal hyperlipidemia, indicating that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) can effectively eliminate masses and eliminate dampness and toxins, thereby achieving effective treatment of membranous nephropathy with renal micro-masses and dampness and toxins.

[0086] The Cr level in serum is one of the important detection indicators of glomerular filtration rate, which is mainly excreted from the body through glomerular filtration and is not reabsorbed by the renal tubules. To some extent, it can explain the damage of glomeruli in the kidney. The increase of BUN level in serum can cause organic lesions such as pyelonephritis, interstitial nephritis, primary glomerulonephritis, and cause prerenal oliguria such as massive dehydration, hepatorenal syndrome, decreased renal blood flow and lack of perfusion, thereby accelerating the development of membranous nephropathy, and also leading to various complications such as decreased insulin, metabolic dysfunction, hyperuricemia and hypertension. The experimental results show that the BUN and Cr levels of the test sample group and the control group decreased to varying degrees compared with the model group, indicating that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) can reduce the BUN and Cr levels to protect glomerular damage, improve renal function, reduce the exudation of proteinuria, and thus delay the progression of membranous nephropathy.

[0087] 3.5 Effect of Chinese herbal medicine combination on renal pathological morphology in rats with membranous nephropathy 3.5.1 HE staining to observe the pathological morphology of rats The results are as follows Fig.10 As shown, from Fig.10 It can be seen that the number of glomeruli in the blank group rats was abundant, the structure was clear, the matrix did not proliferate, and there was no obvious pathological morphology and structural changes. The renal tubular structure of the model group rats was unclear, the basement membrane was significantly thickened, there was punctate infiltration of inflammatory cells, focal atrophy was seen in the tubular cortex, and fibroblast proliferation occurred in the glomerular interstitium. At the same time, the model group rats had mesangial cell proliferation, that is, "micro-symptom" syndrome; after 8 weeks of drug intervention, compared with the model group, the renal tubular structure of the medium and low dose groups of the test product was clear, the epithelial cell morphology was regular, and the number of cells in the glomerulus, mesangial cells, and matrix did not show obvious proliferation; individual atrophy was seen in the renal tubular cortex of the high dose group of the test product, the basement membrane was slightly thickened, and the number of glomerular cells and matrix did not show obvious proliferation; the renal tubular of the control group of Tripterygium wilfordii polyglycosides occasionally showed focal atrophy, unclear structure, thickened basement membrane, and focal infiltration of inflammatory cells in the interstitium.

[0088] These results indicate that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided by Example 1, administered at a dosage of 3.75 g / kg / d and 7.5 g / kg / d, can effectively eliminate masses and dampness and toxin accumulation, thereby making the renal tubular structure clear, the epithelial cell morphology regular, and no obvious proliferation of the number of cells, mesangial cells, and matrix in the glomerulus, thereby achieving effective treatment of renal micro-mass and dampness and toxin accumulation type membranous nephropathy.

[0089] 3.5.2 Masson staining to observe the pathological morphology of rats The results are as follows Fig.11 As shown, from Fig.11 It can be seen that the number, size and morphology of glomeruli in the blank group rats were normal, no obvious fibrosis was observed in the glomeruli, and no inflammatory cell infiltration was observed. In the model group rats, a large number of glomeruli were atrophied and reduced, and the basement membrane was thickened. A large number of collagen fiber proliferation and inflammatory cell aggregation were observed in the glomerular interstitium, renal tubules were atrophied, and the epithelial cell cytoplasm was lightly stained. At the same time, mesangial cell proliferation occurred in the model group rats, that is, "micro-symptom" syndrome; after 8 weeks of drug intervention, compared with the model group, no obvious abnormalities in the number, size, fibrosis, interstitial proliferation, mesangial cell proliferation, and inflammatory cell infiltration were observed in the glomeruli of the test sample medium and low dose groups, and no obvious abnormalities in the renal tubules were observed; in the test sample high dose group and the tripterygium polyglycosides tablet control group, the renal tubule structure was unclear, some were atrophied, the epithelial cell cytoplasm was loose, and small focal infiltration of collagen fibers was observed around.

[0090] These results indicate that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided by Example 1, administered at a dosage of 3.75 g / kg / d and 7.5 g / kg / d, can effectively eliminate masses and dampness and toxin accumulation, thereby eliminating obvious abnormalities in the number, fibrosis, interstitial hyperplasia, and inflammatory cell infiltration in the glomeruli, thereby achieving effective treatment of renal micro-masses and dampness and toxin accumulation type membranous nephropathy.

[0091] 3.5.3 PASM staining to observe the pathological morphology of rats The results are as follows Fig.12 As shown, from Fig.12 It can be seen that the blank group rats did not have thickening of the basement membrane. The basement membrane in the glomeruli of the model group rats was thickened and uneven, indicating that there was inflammation in the glomeruli; after 8 weeks of drug intervention, the thickness of the basement membrane of the glomeruli in the test group and the control group of Tripterygium wilfordii polyglycosides was reduced compared with the model group.

[0092] These results indicate that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided by Example 1) has an anti-inflammatory effect and can effectively eliminate glomerular inflammation, thereby reducing the thickness of the glomerular basement membrane.

[0093] Combination Figure 10-12 It can be seen that compared with the blank group, the model group showed significant pathological morphological damage. The renal tissue of the rats showed shrunken glomeruli, diffuse thickening of the capillary walls, fibrosis, collagen fiber proliferation, mesangial cell proliferation, and inflammatory cell aggregation; the basement membrane was thickened, and the podocytes were arranged irregularly. After 8 weeks of drug intervention, compared with the model group, the pathological morphological damage in the test group and the Tripterygium Wilfordii Polyglycosides Tablets control group was alleviated to varying degrees, among which the medium-dose group of the test sample had the most obvious improvement, with fewer abnormalities in the number and size of glomeruli, alleviated fibrosis in the glomeruli, reduced inflammatory cell infiltration, and reduced basement membrane thickening. These results indicate that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided by Example 1) can effectively eliminate glomerular inflammation, can effectively reduce the proliferation of a large number of collagen fibers (mesangial cell proliferation) visible in the interstitium, can effectively reduce pathological morphological damage, thereby achieving effective treatment of membranous nephropathy of renal micro-mass and dampness and toxin accumulation type, especially administration at a dose of 7.5 g / kg / d can achieve better effects of eliminating masses and dampness and toxin accumulation, so that the number and size of glomeruli in the pathological morphological results are less abnormal, the fibrous proliferation in the glomerulus is alleviated, the infiltration of inflammatory cells is reduced, and the thickening of the basement membrane is alleviated.

[0094] 3.6 Effect of Chinese herbal medicine combination on renal ultrastructure in rats with induced membranous nephropathy 3.6.1 Scanning electron microscopy results The scanning electron microscopy results are shown in Table 6. Fig.13 .

[0095] Table 6 Scanning electron microscopic results of the effect of the Chinese herbal medicine composition on the ultrastructure of rats with membranous nephropathy 3.6.2 Transmission electron microscopy results Transmission electron microscopy shows, see Table 7, Figure 14.

[0096] Table 7 Transmission electron microscopy results of the Chinese herbal medicine combination on the ultrastructure of rats with membranous nephropathy Combined with Table 6, Table 7 and Fig.13 , Fig.14 It can be seen that compared with the blank group, the model group showed pathological morphological damage of the model, severe glomerular damage, uneven basement membrane thickness, dense material deposition, and a large number of fusions of podocyte foot processes. After 8 weeks of drug treatment, the glomerular structure of the test sample medium-dose group was slightly damaged, the podocyte morphology was slightly swollen, and the foot process damage was relatively mild. The glomerular structure of the test sample high- and low-dose groups was moderately damaged, the podocytes were slightly wrinkled and collapsed, the intercellular space was widened, and the foot processes were partially swollen and fused, but the comparison model group was alleviated. The results show that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1, administered at a dosage of 7.5g / kg / d, can effectively eliminate masses and eliminate dampness and toxins, thereby alleviating mild damage to the glomerular structure, alleviating slight swelling of podocytes, and alleviating foot process damage, and achieving effective treatment of membranous nephropathy of renal micro-mass and dampness and toxins.

[0097] In addition, the effect of the Chinese herbal composition in Example 1 on coagulation function was also verified. The results showed that after treatment with the Chinese herbal composition, the coagulation function was significantly improved, indicating that the Chinese herbal composition can effectively reduce platelet aggregation and play a role in eliminating lumps and removing congestion.

[0098] This shows that, from the perspective of Western medicine, the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) can significantly reduce the 24hUTP quantification and mALB levels of rats with membranous nephropathy; reduce the levels of Cr, BUN, TC, TG, and LDL-C, and increase the levels of ALB and TP. This shows that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) can alleviate the abnormal manifestations of rats with membranous nephropathy, can significantly reduce the exudation of proteinuria, and has a good therapeutic effect on membranous nephropathy. At the same time, it protects liver function and kidney function.

[0099] From the perspective of traditional Chinese medicine, the above results (especially the pathological morphology under the light microscope and the ultrastructure under the electron microscope) show that the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided in Example 1) can effectively eliminate masses and congestion, and has a clear therapeutic effect on membranous nephropathy with micro-masses in the kidney and excessive dampness and toxins.

[0100] Test Example 2 Paste yield Test subject: the Chinese medicine composition obtained in Example 1.

[0101] Test method: Take the Chinese medicine composition obtained in Example 1, accurately weigh a certain amount of dry powder, and determine its paste yield. The paste yield is the percentage by weight of the paste yield to the dry powder yield.

[0102] Test results: as shown in Table 8 below.

[0103] Table 8 Effect of different extraction time on the yield of paste

[0104] It can be seen from Table 8 that the paste yield of Example 1 is 23.52%.

[0105] Test Example 3 Dioscin Content Test subject: the Chinese medicine composition obtained in Example 1.

[0106] Test method: High performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Rules 0512).

[0107] Instruments and chromatographic conditions: Agilent 1100 HPLC; Elite SinoChrom C 18 Chromatographic column (4.6 mm×250 mm, 5μm); column temperature: room temperature; mobile phase: acetonitrile-water (55:45), flow rate: 1.0mL / min, detection wavelength: 203nm; injection volume: 10μl.

[0108] Preparation of reference solution: Take an appropriate amount of dioscin reference substance, weigh accurately, and add methanol to make a solution containing 0.3 mg per 1 ml.

[0109] Preparation of the test solution: Take the dry powder of the Chinese medicine composition obtained in Example 1, grind it into powder, take about 3g, accurately weigh it, put it in a stoppered conical bottle, accurately add 25mL of 65% ethanol, weigh it, ultrasonically treat it (150W, 40kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 65% ethanol, filter it, and take the filtrate. Weigh and convert it into the content of diosgenin extracted per gram of dry powder as the basis for calculating the extraction efficiency.

[0110] Test results: as shown in Table 9 below.

[0111] Table 9 Effect of different extraction times on dioscin content

[0112] As can be seen from Table 9, the dioscin content of Example 1 is 1.298 mg / g.

[0113] In summary, by adopting the preparation method provided by the present invention (especially the preparation method provided by Example 1), a higher paste yield and dioscin content can be obtained; the Chinese medicine composition provided by the present invention (especially the Chinese medicine composition provided by Example 1) can effectively treat membranous nephropathy, especially membranous nephropathy of the type with renal micro-mass and dampness and toxicity accumulation.

[0114] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A Chinese medicine composition for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type, the raw materials of which include: The mass ratio of Rhizoma Dioscoreae, Pteris cretica and Hedyotis diffusa is 1:0.6:0.

6.

2. The Chinese medicine composition according to claim 1, characterized in that: Calculated by weight, the raw materials include: 360 parts of Psoralea corylifolia, 360 parts of Hedyotis diffusa and 600 parts of Rhizoma Dioscoreae.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that: The dosage form of the medicine is selected from at least one of decoction, powder, granule and capsule.

4. The method for preparing the Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The water decoction extraction process is adopted, wherein during the water decoction, the amount of water added is 10-12 times the amount of all raw materials, the decoction time is 1.5-2h, and the decoction times are 2-3 times.

5. The preparation method according to claim 4, characterized in that: The method also includes concentration, drying and pulverization. The concentration is to concentrate at 60-70° C. to a relative density of 1.23-1.

28. The temperature during drying is 60-65° C. After pulverization, fine powder of 80-100 meshes is obtained.

6. The preparation method according to claim 5, characterized in that: The method further comprises filling the fine powder into capsules, wherein the content of the traditional Chinese medicine composition in each capsule is 0.4-0.6 g.

7. The preparation method according to claim 6, characterized in that: The method also includes further filling the fine powder into capsules, wherein the content of the traditional Chinese medicine composition is 0.5 g per capsule.

8. Use of the Chinese medicine composition according to any one of claims 1 to 3 and / or the Chinese medicine composition obtained by the preparation method according to any one of claims 4 to 7 in medicines for treating renal micro-mass and membranous nephropathy of dampness and toxicity accumulation type.

9. The use according to claim 8, characterized in that: The Chinese medicine composition is administered orally.

10. The use according to claim 9, characterized in that: The dosage of the Chinese medicine composition is 0.075-0.125 g / kg / d.

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