Application of traditional Chinese medicine composition in preparation of medicine for preventing and treating nephrotic syndrome

Abstract

The invention relates to the technical field of traditional Chinese medicines, in particular to application of a traditional Chinese medicine composition in preparation of a medicine for preventing and treating nephrotic syndrome. The traditional Chinese medicine composition comprises sedum sarmentosum, seaweed, oyster, hawthorn, herba siegesbeckiae, schisandra chinensis and polygonum cuspidatum. It is found that the composition can remarkably reduce the content of urine protein, regulate the levels of ALB, Cr and BUN in serum and improve the renal function, and has a good treatment effect on the nephrotic syndrome. The traditional Chinese medicine composition is an existing Chinese patent medicine, medication is safe, and the preparation method is simple, rapid, efficient and easy to operate. The prepared mixture, tablet, capsule, pellet and the like are convenient to carry and take.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to application of a traditional Chinese medicine composition in preparing medicines for preventing and treating nephrotic syndrome. Background Art

[0002] Nephrotic syndrome (NS) is a group of clinical syndromes characterized by massive proteinuria, hypoproteinemia, severe edema, and hyperlipidemia. It can be caused by a variety of causes and is divided into three categories: primary, secondary, and hereditary. Primary nephrotic syndrome is a primary glomerular disease and consists of a variety of pathological types.

[0003] Nephrotic syndrome is often caused by colds and fatigue. It can occur at any age. Its symptoms are obvious, so it is easy for patients to detect. Generally, there will be a large amount of proteinuria (24-hour urine protein quantity ≥ 3.5 grams), severe edema (in addition to eyelid swelling and lower limb edema, there will also be pleural effusion, ascites, and pericardial effusion), hyperlipidemia, and hypoproteinemia (<30 grams / liter), which are clinically known as the "three highs and one low" symptoms.

[0004] Nephrotic syndrome takes a long time to develop and is difficult to cure, which brings great physical and psychological burden to patients. The current medical level has no obvious special treatment for nephrotic syndrome. The existing treatment drugs have a long course of treatment, a low cure rate, and large side effects, which increase the pain of patients. At present, the incidence of nephrotic syndrome is increasing year by year, and early prevention and treatment to delay its development are increasingly valued by people. Traditional Chinese medicine has been passed down in my country for thousands of years. Chinese herbal compound preparations pay attention to syndrome differentiation and treatment in the treatment of nephrotic syndrome, and have the effect of treating both the symptoms and the root causes. Further development and application of Chinese herbal prescriptions are expected to achieve excellent results in the prevention and treatment of nephrotic syndrome. Summary of the invention

[0005] In view of this, the technical problem to be solved by the present invention is to provide an application of a Chinese medicine composition in the preparation of a medicine for preventing and treating nephrotic syndrome.

[0006] The invention provides application of a traditional Chinese medicine composition in preparing medicine for preventing and treating nephrotic syndrome.

[0007] In the present invention, the Chinese medicine composition comprises the following raw materials in parts by weight: 1-1000 parts of Creeping Herb, 1-500 parts of Seaweed, 1-1000 parts of Oyster, 1-500 parts of Hawthorn, 1-1000 parts of Siegesbeckia, 1-500 parts of Schisandrae Chinensis, and 1-800 parts of Polygonum Cuspidatum.

[0008] In the present invention, the prevention and treatment include: improving renal function and / or reducing urine protein level.

[0009] Renal function refers to the function of the kidney to excrete metabolic waste in the body and maintain the stability of electrolytes such as sodium, potassium, and calcium and the acid-base balance of the body. Renal function tests usually include the measurement of indicators such as blood creatinine (Cr), blood urea nitrogen (BUN), blood and urine β2-microglobulin, urine albumin (ALB), urine immunoglobulin G, and urine secretory immunoglobulin A. The changes in these indicators can reflect the state of renal function. The results of the embodiments of the present invention show that the administration of the Chinese medicine composition can effectively increase the ALB content in rat serum and reduce the Cr and BUN content, and has a good improvement effect on renal function. That is, the improvement of renal function described in the present invention includes increasing the ALB level and reducing the Cr and / or BUN levels.

[0010] Urine protein refers to protein in urine, and the presence of urine protein usually means that kidney function is impaired. The main characteristic of patients with nephrotic syndrome is that there is a large amount of protein in the urine. The results of the examples of the present invention show that the administration of the Chinese medicine composition can significantly reduce the content of urine protein.

[0011] Nephrotic syndrome can be divided into primary nephrotic syndrome and secondary nephrotic syndrome. The nephrotic syndrome described in the present invention is nephrotic syndrome caused by drugs. The results in the examples show that the Chinese medicine composition has a good therapeutic effect on nephrotic syndrome caused by drugs, and the drugs are doxorubicin and / or puromycin aminonucleoside.

[0012] In the embodiment of the present invention, the mass proportions of the components in the traditional Chinese medicine composition are: 923 parts of Creeping Herb, 308 parts of Seaweed, 923 parts of Oyster, 308 parts of Crataegus Pinnatifida, 923 parts of Siegesbeckia scabra, 308 parts of Schisandra chinensis, and 618 parts of Polygonum cuspidatum.

[0013] In a specific embodiment, the preparation method of the Chinese medicine composition comprises: adding water to reflux extraction of Siegesbeckia sibiricum, Herba Lysimachiae, Seaweed, Oyster, Hawthorn, Polygonum cuspidatum, and Schisandra chinensis, and concentrating and drying the filtrate. In this embodiment, in the reflux extraction, the mass ratio of water to the Chinese medicine composition is (4-12):1, and the reflux extraction is performed 1-3 times, each time for 1-2 hours, and the density of the extract is 1.25-1.30.

[0014] Preferably, the extraction method comprises: taking Siegesbeckia sibiricum, Herba Lysimachiae, seaweed, oyster, hawthorn, Polygonum cuspidatum and Schisandra chinensis, adding 8 times the amount of water, refluxing and extracting twice, each time for 1.5 hours, filtering, decompressing and concentrating to an extract with a relative density of 1.25 to 1.30 (60±5°C), and then concentrating and drying. The drying adopts belt vacuum drying, and then further comprises the steps of crushing, sieving, adding sucralose and an appropriate amount of cyclodextrin, mixing, and granulating.

[0015] Furthermore, the present invention also provides a medicine for preventing and treating nephrotic syndrome, which includes pharmaceutically acceptable excipients and the following raw materials in parts by weight: 1-1000 g of Creeping Peony, 1-500 g of seaweed, 1-1000 g of oyster, 1-500 g of hawthorn, 1-1000 g of Siegesbeckia, 1-500 g of Schisandra chinensis, and 1-800 g of Polygonum cuspidatum.

[0016] The dosage form of the drug of the present invention includes but is not limited to decoction, granules, capsules, tablets, oral liquids, pills, tinctures, syrups, suppositories, gels, sprays and / or injections.

[0017] In some embodiments provided by the present invention, the capsule is a hard capsule or a soft capsule.

[0018] In some embodiments provided herein, the tablet is an oral tablet or an oral tablet.

[0019] Oral tablets refer to tablets for oral administration. Most of the drugs in such tablets are absorbed through the gastrointestinal tract to exert their effects, and some of the drugs in tablets exert their effects locally in the gastrointestinal tract. In some embodiments provided by the present invention, the oral tablets are ordinary compressed tablets, dispersible tablets, effervescent tablets, chewable tablets, coated tablets or sustained-release tablets.

[0020] The pharmaceutically acceptable excipients include one or a mixture of two or more of fruit powder, edible flavors, sweeteners, acidulants, fillers, lubricants, preservatives, suspending agents, edible pigments, diluents, emulsifiers, disintegrants or plasticizers.

[0021] The medicine described in the present invention also includes other medicines for treating nephrotic syndrome, and the other medicines for treating nephrotic syndrome include: at least one of benazepril, prednisone, dexamethasone, furosemide, hydrochlorothiazide, cyclophosphamide, cyclosporine A, and tacrolimus.

[0022] In the present invention, in the medicine, the Chinese medicine composition and other therapeutic agents may exist independently of each other or may exist in combination, and the present invention does not limit this.

[0023] Furthermore, the present invention also provides a method for preventing and treating nephrotic syndrome, which comprises administering the drug of the present invention.

[0024] The administration of the drug includes, but is not limited to, oral administration, spray inhalation, rectal administration, nasal administration, buccal administration, vaginal administration, topical administration. Or non-enteral administration, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion; or administration via an explanted reservoir. Preferably, the administration is oral, intraperitoneal or intravenous.

[0025] In the present invention, when the medicine includes a Chinese medicine composition and other therapeutic agents, two or more medicines can be administered simultaneously or sequentially, and the present invention is not limited to this.

[0026] For example, the Chinese medicine composition in the medicine is administered no longer than 6 hours, preferably no longer than 5 hours, more preferably no longer than 4 hours, even more preferably no longer than 3 hours, more preferably no longer than 2 hours and most preferably no longer than 1 hour before the administration of other therapeutic agents and / or no longer than 6 hours, preferably no longer than 5 hours, more preferably no longer than 4 hours, even more preferably no longer than 3 hours, more preferably no longer than 2 hours and most preferably no longer than 1 hour after the administration of other therapeutic agents.

[0027] In the present invention, when the medicine includes a Chinese medicine composition and other therapeutic agents, the administration frequency of two or more medicines may be the same or different, and the present invention is not limited to this.

[0028] For example, the administration frequency of two or more of the drugs is independently selected from 4 times / day, 3 times / day, 2 times / day or 1 time / day, wherein the administration frequency of the Chinese medicine composition is 4 times / day, 3 times / day, 2 times / day or 1 time / day.

[0029] In the present invention, the medicine includes a Chinese medicine composition, and the dosage is not less than the effective dose. If the medicine includes a Chinese medicine composition and other therapeutic agents, the dosage of the Chinese medicine composition is lower than the effective dose, or not lower than the effective dose; the dosage of other therapeutic agents is lower than the effective dose, or not lower than the effective dose.

[0030] In the present invention, the effective dose includes a "therapeutically effective amount" or a "prophylactically effective amount", which refers to an amount that is sufficient to treat or prevent a patient's disease but low enough to avoid serious side effects (at a reasonable benefit / risk ratio) within the scope of reasonable medical judgment. The preventive or therapeutic effective amount of a drug will vary depending on the specific drug selected (e.g., considering the efficacy, effectiveness and half-life of the drug), the selected route of administration, the disease to be prevented or treated, the severity of the disease to be prevented or treated, the age, size, weight and physical illness of the patient to be prevented or treated, the medical history of the patient to be prevented or treated, the duration of prevention or treatment, the nature of concurrent therapy, the desired preventive or therapeutic effect, and other factors, but can still be routinely determined by those skilled in the art.

[0031] In the present invention, the control includes prevention and / or treatment, which is a preventive and therapeutic measure for nephrotic syndrome. This includes a method for preventing the occurrence of nephrotic syndrome, the purpose of which is to prevent, slow down (reduce) undesirable physiological changes or lesions in the treatment object, and the beneficial or desired clinical results include but are not limited to avoiding the occurrence of nephrotic syndrome, the alleviation of nephrotic syndrome symptoms, the weakening of nephrotic syndrome, the stabilization of nephrotic syndrome state (i.e., no deterioration), the delay or slowing of nephrotic syndrome progress, the improvement or alleviation of nephrotic syndrome state, and relief (whether partial relief or complete relief), whether detectable or undetectable. The object to be treated includes an object already suffering from a disease or disease and an object susceptible to a disease or disease or an object intended to prevent a disease or disease.

[0032] In the present invention, the object of prevention and treatment is a human or a mammal, and the mammal includes bovines, equines, ovines, porcines, canines, felines, rodents, and primates.

[0033] The Chinese medicine composition of the present invention comprises Herba Lycopodii, seaweed, oyster, hawthorn, herba Siegesbeckiae, schisandrae chinensis and polygonum cuspidatum. The present invention finds that the composition can significantly reduce urine protein content, regulate serum ALB, Cr, BUN levels, improve renal function, and has a good therapeutic effect on nephrotic syndrome. Moreover, the Chinese medicine combination is an existing Chinese patent medicine, safe for use, and the preparation method is simple, fast, efficient, and easy to operate. The prepared mixture, tablets, capsules, pellets, etc. are easy to carry and convenient to take. DETAILED DESCRIPTION

[0034] The present invention provides the application of Chinese medicine composition in the preparation of medicine for preventing and treating nephrotic syndrome, and those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described by preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application of this article without departing from the content, spirit and scope of the present invention to realize and apply the technology of the present invention.

[0035] The present invention proposes an application of a Chinese medicine composition in the preparation of a drug for treating nephrotic syndrome, wherein the Chinese medicine composition comprises: 1 to 1000 parts of Herba Creepersii, 1 to 500 parts of Seaweed, 1 to 1000 parts of Oyster, 1 to 500 parts of Crataegus, 1 to 1000 parts of Siegesbeckia, 1 to 500 parts of Schisandrae Chinensis, and 1 to 800 parts of Polygonum cuspidatum. . The use may be any method that is beneficial to improving the corresponding symptoms of the patient, including treatment or prevention. The composition of the present invention may be directly ground into powder, or may be an extract prepared by conventional means in the art. The Chinese medicine used in the composition of the present invention may also be used in the form of direct grinding into powder, extract, or other processed forms.

[0036] Furthermore, the Chinese medicine composition includes: 1-1000 parts of Creeping Jasmine, 1-500 parts of seaweed, 1-1000 parts of oyster, 1-500 parts of hawthorn, 1-1000 parts of Siegesbeckia, 1-500 parts of Schisandra chinensis, and 1-800 parts of Polygonum cuspidatum.

[0037] Furthermore, the Chinese medicine composition comprises: 500-1000 parts of Herba Creepers, 200-400 parts of Seaweed, 500-1000 parts of Oyster, 200-400 parts of Hawthorn, 500-1000 parts of Siegesbeckia, 200-400 parts of Schisandrae Chinensis, 300-700 parts of Polygonum Cuspidatum

[0038] Preferably, the Chinese medicine composition comprises: 700-950 parts of Creeping Jasmine, 200-350 parts of seaweed, 700-950 parts of oysters, 200-350 parts of hawthorn, 700-950 parts of Siegesbeckia, 200-350 parts of Schisandra chinensis, and 500-650 parts of Polygonum cuspidatum.

[0039] Most preferably, the Chinese medicine composition comprises: 923 parts of Siegesbeckia serrata, 923 parts of Creeping Herb, 308 parts of Sargassum, 923 parts of Oyster, 308 parts of Crataegus pinnatifida, 618 parts of Polygonum cuspidatum, and 308 parts of Schisandra chinensis.

[0040] Specifically, the drug for treating nephrotic syndrome is selected from oral dosage forms, injection dosage forms or external administration preparations.

[0041] Specifically, the drug for treating nephrotic syndrome is selected from decoctions, tablets, capsules, granules, pills, injections, decoctions, suspensions, dispersions, syrups, suppositories, gels, aerosols, patches, and oral liquids.

[0042] Among them, the composition of the present invention described above can be directly ground into powder by traditional methods, or it can be obtained by conventional means to obtain extracts so as to be made into different dosage forms, obtain better therapeutic effects and be more conducive to the preparation of modern pharmaceutical dosage forms.

[0043] The present invention also provides a method for preparing any of the aforementioned Chinese medicine compositions, the preparation method comprising:

[0044] Add water to reflux extraction of Siegesbeckia scabra, Creeping Herb, Sargassum, Calcined Oyster, Crataegus pinnatifida, Polygonum cuspidatum and Schisandra chinensis, adding 8 times the amount of water each time, extracting for 1.5 hours, filtering, and concentrating the filtrate under reduced pressure at no higher than 80°C to an extract with a relative density of 1.25-1.30 (60±5°C), belt vacuum drying, crushing and sieving, adding sucralose and dextrin to mix well, and obtaining granules.

[0045] The compound combination medicine for treating nephrotic syndrome of the present invention can significantly reduce the urine protein content of rats with nephrotic syndrome, increase the ALB content in rat serum and reduce the Cr and BUN content, and improve the renal function of rats with nephrotic syndrome. In addition, the preparation method of the compound combination medicine for treating nephrotic syndrome is simple, fast, efficient and easy to operate. The prepared mixture, tablets, capsules, micropills, etc. are easy to carry and convenient to take.

[0046] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0047] The following experiments, if no specific conditions are specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The APIs or excipients, reagents or instruments used, if the manufacturer is not specified, are all conventional products that can be purchased commercially. Unless otherwise specified, all percentages, ratios, proportions or parts are by weight.

[0048] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention. The present invention is further described below in conjunction with the embodiments:

[0049] Example 1 Preparation of Chinese medicine composition

[0050] Seven herbs in total, 923 g of Siegesbeckia scabra, 923 g of Creeping Herb, 308 g of Sargassum, 923 g of Oyster, 308 g of Crataegus pinnatifida, 618 g of Polygonum cuspidatum and 308 g of Schisandra chinensis, were extracted twice with water under reflux, with 8 times the amount of water added each time. The extraction lasted for 1.5 hours and the extract was filtered. The filtrate was concentrated under reduced pressure at a temperature not higher than 80°C to an extract with a relative density of 1.25-1.30 (60±5°C). The extract was dried under belt vacuum, crushed and sieved to obtain a composite intermediate. 2 g of sucralose and an appropriate amount of dextrin were added and mixed to obtain 1000 g of granules.

[0051] Experimental Example 2 Therapeutic effect of Chinese medicine composition on adriamycin-induced nephrotic syndrome in rats

[0052] 1. Experimental Materials

[0053] 1.1 Animals

[0054] Wistar rats, male, SPF grade, 60, weighing 180-200 g, were purchased from Qinglongshan Animal Breeding Farm, Jiangning District, Nanjing.

[0055] Breeding environment: room temperature 20-26°C, relative humidity controlled at 40%-70%, animal breeding density 5 per cage.

[0056] 1.2 Drugs

[0057] The intermediate of the composition prepared in Example 1, Jiangsu Kangyuan Pharmaceutical Co., Ltd.; Benazepril tablets (No. 2007, Beijing Novartis Pharmaceutical Co., Ltd.); doxorubicin hydrochloride (Sigma, USA, batch number 25316-40-9); pentobarbital sodium (Beijing Chemical Reagent Company, batch number: 211122); physiological saline (Chenxin Pharmaceutical Co., Ltd., batch number: 2101282721); rat urine protein (UP) ELISA kit (Shanghai Ruifan Biological, item number: RF10966); rat ALBelisa kit (Nanjing Jiancheng Bioengineering Institute, item number H021-1-2), rat Cr ELISA kit (Nanjing Jiancheng Bioengineering Institute, H035-1-2), rat BUN ELISA kit (Nanjing Jiancheng Bioengineering Institute, H032-1-2).

[0058] 1.3 Instruments

[0059] Refrigerator (Haier Company, model: BCD-216SZ); electronic balance (Sartorius Scientific Instruments Company, model: BSA224S-CW); purified water instrument (Millipore Company, Germany, model: Milli-QPlus); animal weight scale (Shanghai Xiangchuan Electronic Scale Co., Ltd., model: CX-SC-DA); multi-function microplate reader (MD Company, USA, model: Flexstation 3).

[0060] 2. Dosage design

[0061] The daily dosage of the Chinese medicine composition is 8.62 g of crude drug / day. According to the body surface area conversion method, the equivalent dose for rats is 8.62 g / 60 kg × 6.2 = 0.89 g of crude drug / kg. According to the 1 / 4:1 / 2:1 dosing method, the low dose is 0.22 g of crude drug / kg, the medium dose is 0.45 g of crude drug, and the high dose is 0.89 g. Oral administration, 0.1 mL / 10 g, once a day.

[0062] The daily dosage of benazepril tablets is 10 mg / day. According to the body surface area conversion method, the equivalent dose for rats is 10 mg / 60 kg×6.3=1.1 mg / kg.

[0063] 3. Experimental Methods

[0064] Wistar rats were fed adaptively for 3 days and were free to eat feed and water. After the urine protein test paper method detected no abnormal urine protein, the remaining rats except the normal control group were injected with doxorubicin hydrochloride (6 mg / kg) once according to their body weight to replicate the rat doxorubicin nephrotic syndrome model. The urine protein content of each rat was measured to determine whether the model was successfully replicated. The rats with successful modeling were selected and randomly divided into a model control group, a benazepril tablet group, and a high-, medium-, and low-dose group of the Chinese medicine composition according to the urine protein content. After the animals were grouped, they were gavaged and administered once a day for 2 consecutive weeks. The dosage of each group: benazepril tablet group (1.1 mg / kg), low-, medium-, and high-dose Chinese medicine composition (0.22 g / kg, 0.45 g / kg, 0.89 g / kg).

[0065] During the 2-week administration period, rats in each group were placed in a clean metabolic cage once a week to collect 24-hour urine, and the 24-hour urine protein excretion was determined by the biuret method. After the experiment, the rats were anesthetized by sodium pentobarbital injection, and blood was collected by puncturing the abdominal aorta with a syringe. After standing at room temperature for 3 hours, the serum was separated and stored at -20°C for testing.

[0066] The detection indicators include: urine protein content detection; serum ALB, Cr, BUN biochemical index detection.

[0067] 4. Statistical methods

[0068] The results were statistically analyzed using t-test.

[0069] 5. Experimental results

[0070] 5.1 Effects on urine protein content in rats with adriamycin-induced nephrotic syndrome

[0071] Compared with the normal control group, the urine protein content of the model group rats in the first and second weeks of administration was significantly increased (P<0.001), indicating that the model was successfully established. Compared with the model group, the urine protein content of the Benazepril tablet group was significantly reduced (P<0.001), and the urine protein content of the medium and high dose groups of the Chinese medicine composition was significantly reduced (P<0.01), indicating that the Chinese medicine composition has a good improvement effect on adriamycin-induced renal injury in rats. The results are shown in Table 1.

[0072] Table 1 Effect of adriamycin on urine protein content in rats with nephrotic syndrome induced by adriamycin

[0073]

[0074] Note: Compared with the normal control group, ###P<0.001; compared with the model control group, *P<0.05, **P<0.01.

[0075] 5.2 Effects on serum biochemical parameters in rats with adriamycin-induced nephrotic syndrome

[0076] Compared with the normal control group, the ALB, Cr, and BUN in the serum of the model group rats were significantly increased (P<0.01); after treatment with Benazepril Tablets, compared with the model control group, the ALB in the serum of the Benazepril Tablets group animals increased significantly (P<0.01), and Cr and BUN decreased significantly (P<0.01); after treatment with the Chinese medicine composition, the medium and high dose groups could also significantly increase the ALB content in the serum of rats and reduce the Cr and BUN content (P<0.05, P<0.01). The results are shown in Table 2.

[0077] Table 2 Effects of serum biochemical parameters on adriamycin-induced nephrotic syndrome in rats ( n=8)

[0078]

[0079] Note: Compared with the normal control group, ##P<0.01; compared with the model control group, **P<0.01, *P<0.05.

[0080] 6. Experimental conclusion

[0081] The Chinese herbal composition at 0.45 and 0.89 g crude drug / kg can significantly regulate the urine protein content and serum ALB, Cr, and BUN biochemical indicators of rats with doxorubicin-induced nephrotic syndrome, indicating that the Chinese herbal composition has a certain improvement effect on the renal function of rats with doxorubicin-induced nephrotic syndrome.

[0082] Experimental Example 3 Therapeutic effect of Chinese medicine composition on puromycin aminonucleoside-induced nephrotic syndrome rat model

[0083] 1. Experimental Materials

[0084] 1.1 Animals

[0085] 60 SD rats, male, SPF grade, weighing 250-270 g, were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd.

[0086] Breeding environment: room temperature 20-26℃, relative humidity controlled at 40%-70%, and breeding density of 5 per cage.

[0087] 1.2 Drugs

[0088] The Chinese medicine composition was prepared according to the method of Example 1 and provided by Jiangsu Kangyuan Pharmaceutical Co., Ltd.; benazepril tablets (No. 2007, Beijing Novartis Pharmaceutical Co., Ltd.); puromycin aminonucleoside (PAN), purchased from Sigma (Lot No. 096K4012); chloral hydrate (Sinopharm Chemical Reagent Co., Ltd., batch number 20200320); rat urine protein (UP) ELISA kit (Shanghai Ruifan Biological, item number: RF10966); rat ALB ELISA kit (Nanjing Jiancheng Bioengineering Institute, item number H021-1-2), rat Cr ELISA kit (Nanjing Jiancheng Bioengineering Institute, H035-1-2), rat BUN ELISA kit (Nanjing Jiancheng Bioengineering Institute, H032-1-2); normal saline (Chenxin Pharmaceutical Co., Ltd., batch number 2101282721).

[0089] 1.3 Instruments

[0090] Electronic balance (Sartorius Scientific Instruments, model BS224S); low-temperature centrifuge (eppendorf, Germany, model 5804R); purified water instrument (Millipore, Germany, model: Milli-Q Plus); animal weight scale (Shanghai Xiangchuan Electronic Scale Co., Ltd., model: CX-SC-DA); ELISA reader (MeiGu Molecular, USA, model Flexstation3).

[0091] 2. Dosage design

[0092] The daily dosage of the Chinese medicine composition is 8.62 g of crude drug / day. According to the body surface area conversion method, the equivalent dose for rats is 8.62 g / 60 kg × 6.2 = 0.89 g of crude drug / kg. According to the 1 / 4:1 / 2:1 dosing method, the low dose is 0.22 g of crude drug / kg, the medium dose is 0.45 g / crude drug, and the high dose is 0.89 g / crude drug. Oral administration, 0.1 mL / 10 g, once a day.

[0093] The daily dosage of benazepril tablets is 10 mg / day. According to the body surface area conversion method, the equivalent dose for rats is 10 mg / 60 kg×6.3=1.1 mg / kg.

[0094] 3. Experimental Methods

[0095] 60 Wistar rats were fed adaptively for 3 days and were free to eat feed and water. After urine protein test paper method detected no abnormal urine protein, they were divided into 6 groups by random number table method, namely normal control group, model control group, dexamethasone control group, high, medium and low dose groups of Chinese medicine composition, with 10 rats in each group. After successful intraperitoneal anesthesia with 10% chloral hydrate (0.33g / kg body weight), the rats were fixed in supine position, the neck was prepared, and 75% alcohol was disinfected. Except for the normal control group, rats in each group underwent jugular vein cannulation and slowly injected PAN saline solution 40mg / kg body weight. The control group underwent jugular vein cannulation and slowly injected the same volume of 0.9% saline. Rats in each group were placed in clean metabolic cages on the 5th day after jugular vein cannulation, fasted and given water, and 24h urine was collected. The 24h urine protein excretion was determined by the biuret method. Compared with the blank group, a significant increase in urine protein was used as the criterion for successful modeling. At the same time, from the 6th day of the experiment, each group began intraperitoneal injection and oral gavage administration, benazepril tablets group (1.1 mg / kg), low, medium and high doses of Chinese medicine combination (0.22 g / kg, 0.45 g / kg, 0.89 g / kg), once a day for 2 consecutive weeks.

[0096] During the 2-week administration period, rats in each group were placed in a clean metabolic cage once a week to collect 24-hour urine, and the 24-hour urine protein excretion was determined by the biuret method. After the experiment, the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate, and blood was collected by puncturing the abdominal aorta with a syringe. After standing at room temperature for 3 hours, the serum was separated and stored at -20°C for testing.

[0097] The detection indicators are: 1) weekly urine protein content detection; 2) serum ALB, Cr, BUN biochemical index detection at the experimental endpoint.

[0098] 4. Statistical methods

[0099] The results were statistically analyzed using t-test.

[0100] 5. Experimental results

[0101] 5.1 Effects on urine protein content in rats with nephrotic syndrome induced by puromycin aminonucleoside

[0102] Compared with the normal control group, the urine protein content of the model group rats in the first and second weeks of administration was significantly increased (P<0.001); compared with the model group, the urine protein content of the Benazepril tablet group was significantly reduced (P<0.001), and the urine protein content of the medium and high dose groups of the Chinese medicine composition was significantly reduced (P<0.01). The results are shown in Table 3.

[0103] Table 3 Effects on urine protein content in rats with nephrotic syndrome induced by puromycin aminonucleoside

[0104]

[0105] Note: Compared with the normal control group, ###P<0.001; compared with the model control group, *P<0.05, **P<0.01.

[0106] 5.2 Effects on serum biochemical indices of rats with nephrotic syndrome induced by puromycin aminonucleoside Compared with the normal control group, the serum ALB, Cr, and BUN of the model group rats were significantly increased (P<0.001); after treatment with benazepril tablets, the serum ALB, Cr, and BUN of the benazepril tablets group animals were significantly reduced compared with the model control group (P<0.001); the medium and high dose groups of the Chinese medicine composition can significantly increase the ALB content in the serum of rats (P<0.01, P<0.001), and significantly reduce the Cr and BUN content (P<0.01, P<0.001). The results are shown in Table 4.

[0107] Table 4 Effects on serum biochemical parameters in rats with nephrotic syndrome induced by puromycin aminonucleoside

[0108]

[0109] Note: Compared with the normal control group, ###P<0.001; compared with the model control group, ***P<0.001, **P<0.01

[0110] 6. Experimental conclusion

[0111] The Chinese medicine composition at 0.45 and 0.89 g / kg can significantly reduce the urine protein content in the puromycin aminonucleoside-induced nephrotic syndrome rat model, and significantly increase the serum ALB content of the rats and reduce the Cr and BUN content, indicating that the Chinese medicine composition has the effect of alleviating and improving kidney damage in nephrotic syndrome rats, and the Chinese medicine composition has a certain therapeutic effect on the puromycin aminonucleoside-induced nephrotic syndrome rat model.

[0112] The above are only preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of a Chinese medicine composition in the preparation of a drug for preventing and treating nephrotic syndrome; The Chinese medicine composition comprises the following raw materials in parts by weight: 1-1000 parts of Creeping Herb, 1-500 parts of seaweed, 1-1000 parts of oyster, 1-500 parts of hawthorn, 1-1000 parts of Siegesbeckia, 1-500 parts of Schisandra chinensis, and 1-800 parts of Polygonum cuspidatum.

2. The use according to claim 1, characterized in that: The prevention and treatment include: improving renal function and / or reducing urine protein level.

3. The use according to claim 2, characterized in that: The improvement of renal function includes increasing ALB level and decreasing Cr and / or BUN level.

4. The use according to claim 1, characterized in that: The nephrotic syndrome is drug-induced nephrotic syndrome.

5. The use according to claim 4, characterized in that: The drug is doxorubicin and / or puromycin aminonucleoside.

6. The use according to any one of claims 1 to 5, characterized in that: The preparation method of the traditional Chinese medicine composition comprises: adding water to extract siegesbeckia, scutellaria baicalensis, seaweed, oyster, hawthorn, knotweed and schisandra chinensis, adding 5-10 times the mass of water each time, extracting for 2-4 hours, filtering, concentrating the filtrate under reduced pressure at a temperature not higher than 80° C. to an extract with a relative density of 1.25-1.30 (60±5° C.), and then concentrating and drying.

7. The use according to claim 67, characterized in that In the reflux extraction, reflux extraction was performed twice, each time for 1.5 hours.

8. A drug for preventing and treating nephrotic syndrome, comprising pharmaceutically acceptable excipients and the following raw materials in parts by weight: 1-1000 parts of Creeping Peony vine, 1-500 parts of seaweed, 1-1000 parts of oyster, 1-500 parts of hawthorn, 1-1000 parts of Siegesbeckia scabra, 1-500 parts of Schisandra chinensis, and 1-800 parts of Polygonum cuspidatum.

9. The drug according to claim 8, characterized in that Other drugs for treating nephrotic syndrome are also included, and the other drugs for treating nephrotic syndrome include: at least one of benazepril, prednisone, dexamethasone, furosemide, hydrochlorothiazide, cyclophosphamide, cyclosporine A, and tacrolimus.

Images