Application of Traditional Chinese Medicine Compositions in the Preparation of Drugs for the Prevention and Treatment of Nephrotic Syndrome

Abstract

This invention relates to the field of traditional Chinese medicine (TCM) technology, and particularly to the application of TCM compositions in the preparation of drugs for the prevention and treatment of nephrotic syndrome. The TCM composition of this invention includes *Sedum sarmentosum*, seaweed, oyster shell, hawthorn, *Siegesbeckia orientalis*, *Schisandra chinensis*, and *Polygonum cuspidatum*. This invention has found that this composition can significantly reduce urinary protein levels, regulate serum ALB, Cr, and BUN levels, improve renal function, and has a good therapeutic effect on nephrotic syndrome. Moreover, this TCM composition is an existing prepared Chinese medicine, safe to use, and its preparation method is simple, rapid, efficient, and easy to operate. The prepared mixtures, tablets, capsules, and micro-pellets are easy to carry and convenient to take.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to the application of traditional Chinese medicine compositions in the preparation of drugs for the prevention and treatment of nephrotic syndrome. Background Technology

[0002] Nephrotic syndrome (NS) is a group of clinical symptoms characterized by massive proteinuria, hypoalbuminemia, severe edema, and hyperlipidemia. It can be caused by a variety of etiologies and is divided into three main categories: primary, secondary, and hereditary. Primary nephrotic syndrome belongs to primary glomerular diseases and has multiple pathological types.

[0003] Nephrotic syndrome often begins with a cold or overexertion and can occur at any age. Its symptoms are obvious, making it easy for patients to notice. Common symptoms include massive proteinuria (24-hour urine protein ≥3.5 g), severe edema (in addition to eyelid and lower extremity edema, pleural effusion, ascites, and pericardial effusion), hyperlipidemia, and hypoproteinemia (<30 g / L), clinically known as the "three highs and one low" syndrome.

[0004] Nephrotic syndrome is a chronic condition that is difficult to cure, placing a significant physical and psychological burden on patients. Current medical technology lacks a highly effective treatment for nephrotic syndrome, and existing medications often involve long treatment courses, low cure rates, and significant side effects, further increasing patient suffering. The incidence of nephrotic syndrome is increasing annually, making early prevention and slowing its progression increasingly important. Traditional Chinese medicine (TCM), with its thousands of years of history, emphasizes syndrome differentiation and treatment in the management of nephrotic syndrome, offering both symptomatic relief and root cause treatment. Further development and application of TCM formulas 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 the application of traditional Chinese medicine composition in the preparation of drugs for the prevention and treatment of nephrotic syndrome.

[0006] This invention provides the application of a traditional Chinese medicine composition in the preparation of a drug for the prevention and treatment of nephrotic syndrome.

[0007] In this invention, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 1-1000 parts of Sedum sarmentosum, 1-500 parts of Sargassum, 1-1000 parts of Oyster shell, 1-500 parts of Crataegus pinnatifida, 1-1000 parts of Siegesbeckia orientalis, 1-500 parts of Schisandra chinensis, and 1-800 parts of Polygonum cuspidatum.

[0008] In this invention, the prevention and treatment include: improving renal function and / or reducing urinary protein levels.

[0009] Kidney function refers to the kidneys' ability to excrete metabolic waste products and maintain the stability of electrolytes such as sodium, potassium, and calcium, as well as acid-base balance. Kidney function tests typically include measurements of serum creatinine (Cr), blood urea nitrogen (BUN), serum and urinary β2-microglobulin, urinary albumin (ALB), urinary immunoglobulin G, and urinary secretory immunoglobulin A. Changes in these indicators reflect the state of kidney function. The results of the embodiments of this invention show that administration of the aforementioned traditional Chinese medicine composition can effectively increase the ALB content in rat serum and decrease the Cr and BUN levels, exhibiting a good improving effect on kidney function. That is, the improvement of kidney function described in this invention includes increasing ALB levels and decreasing Cr and / or BUN levels.

[0010] Proteinuria refers to the presence of protein in urine, and its presence usually indicates impaired kidney function. A key characteristic of nephrotic syndrome is the presence of high levels of protein in the urine. The results of this invention's embodiments demonstrate that administering the aforementioned traditional Chinese medicine composition can significantly reduce proteinuria levels.

[0011] Nephrotic syndrome can be divided into primary nephrotic syndrome and secondary nephrotic syndrome. The nephrotic syndrome described in this invention is drug-induced nephrotic syndrome. Results from the examples show that the traditional Chinese medicine composition has a good therapeutic effect on drug-induced nephrotic syndrome, wherein the drugs are doxorubicin and / or puromycin aminonucleoside.

[0012] In this embodiment of the invention, the mass fractions of each component in the traditional Chinese medicine composition are as follows: 923 parts of Sedum sarmentosum, 308 parts of Sargassum, 923 parts of Ostrea gigas, 308 parts of Crataegus pinnatifida, 923 parts of Siegesbeckia orientalis, 308 parts of Schisandra chinensis, and 618 parts of Polygonum cuspidatum.

[0013] In a specific embodiment, the preparation method of the traditional Chinese medicine composition includes: extracting Siegesbeckia orientalis, Sedum sarmentosum, Sargassum, Ostrea gigas, Crataegus pinnatifida, Polygonum cuspidatum, and Schisandra chinensis by reflux with water, and concentrating and drying the filtrate. In this embodiment, the mass ratio of water to the traditional Chinese medicine composition in the reflux extraction is (4-12):1, 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] As a preferred method, the extraction method includes: taking Siegesbeckia orientalis, Sedum sarmentosum, seaweed, oyster shell, hawthorn, Polygonum cuspidatum, and Schisandra chinensis, adding 8 times the amount of water, reflux extraction twice, 1.5 hours each time, filtering, concentrating under reduced pressure to an extract with a relative density of 1.25-1.30 (60±5℃), and then concentrating and drying. The drying is carried out using a belt vacuum dryer, and the process also includes pulverizing, sieving, adding sucralose and an appropriate amount of cyclodextrin, mixing, and granulating.

[0015] Furthermore, the present invention also provides a drug for the prevention and treatment of nephrotic syndrome, comprising pharmaceutically acceptable excipients and the following raw materials in parts by weight: 1-1000g of Sedum sarmentosum, 1-500g of Sargassum, 1-1000g of Ostrea gigas, 1-500g of Crataegus pinnatifida, 1-1000g of Siegesbeckia orientalis, 1-500g of Schisandra chinensis, and 1-800g of Polygonum cuspidatum.

[0016] The dosage forms of the drugs described in this invention include, but are not limited to, decoctions, 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 by the present invention, the tablet is an oral tablet or a dental tablet.

[0019] Oral tablets refer to tablets intended for oral administration. Most of these tablets contain drugs that are absorbed through the gastrointestinal tract to exert their effects, while some contain drugs that exert their effects locally within the gastrointestinal tract. In some embodiments provided by this 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 more of the following: fruit powder, flavoring, sweetener, acidulant, filler, lubricant, preservative, suspending agent, food coloring, diluent, emulsifier, disintegrant, or plasticizer.

[0021] The medicament described in this invention also includes other medicaments for treating nephrotic syndrome, including at least one of benazepril, prednisone, dexamethasone, furosemide, hydrochlorothiazide, cyclophosphamide, cyclosporine A, and tacrolimus.

[0022] In this invention, the traditional Chinese medicine composition and other therapeutic agents in the drug can exist independently or in combination, and this invention does not limit this.

[0023] Furthermore, the present invention also provides a method for preventing and treating nephrotic syndrome, comprising administering the drug described herein.

[0024] The administration methods of the drug include, but are not limited to, oral, spray inhalation, rectal, nasal, buccal, vaginal, and topical administration. Alternatively, it may be administered via non-enteric routes, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal, or intracranial injection or infusion; or via an external implantation device. Preferably, the administration method is oral, intraperitoneal, or intravenous administration.

[0025] In this invention, when the drug includes a traditional Chinese medicine composition and other therapeutic agents, two or more drugs may be administered simultaneously or sequentially, and this invention does not limit this.

[0026] For example, the traditional Chinese medicine composition in the drug is applied no longer than 6 hours before the application of other therapeutic agents, 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 and / or no longer than 6 hours after the application of other therapeutic agents, 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.

[0027] In this invention, when the drug includes a traditional Chinese medicine composition and other therapeutic agents, the dosing frequencies of two or more drugs may be the same or different, and this invention does not limit this.

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

[0029] In this invention, the drug comprises a traditional Chinese medicine composition, and the dosage is not less than the effective dose. If the drug comprises a traditional Chinese medicine composition and other therapeutic agents, the dosage of the traditional Chinese medicine composition is either less than or not less than the effective dose; the dosage of the other therapeutic agents is either less than or not less than the effective dose.

[0030] In this invention, the effective dose includes a "therapeutic effective dose" or a "preventive effective dose," which refers to an amount sufficient, within reasonable medical judgment, to treat or prevent a patient's disease while avoiding serious side effects with a sufficiently low risk-benefit ratio. The preventive or therapeutic effective dose of a drug will vary depending on factors such as the specific drug chosen (e.g., considering the drug's potency, effectiveness, and half-life), the chosen route of administration, the disease being prevented or treated, the severity of the disease being prevented or treated, the patient's age, size, weight, and physical condition, the patient's medical history, the duration of prevention or treatment, the nature of concurrent therapies, and the desired preventive or therapeutic effect, but can still be routinely determined by those skilled in the art.

[0031] In this invention, the prevention and / or treatment encompasses preventive and / or therapeutic measures for nephrotic syndrome. This includes methods for preventing the occurrence of nephrotic syndrome, the purpose of which is to prevent, mitigate (reduce) undesirable physiological changes or lesions in the treated subject. Beneficial or desired clinical outcomes include, but are not limited to, avoidance of nephrotic syndrome, reduction of nephrotic syndrome symptoms, decrease in the severity of nephrotic syndrome, stabilization of the nephrotic syndrome state (i.e., no worsening), delay or slowing of the progression of nephrotic syndrome, improvement or mitigation of the nephrotic syndrome state, and remission (whether partial or complete remission), whether detectable or undetectable. Subjects requiring treatment include those already suffering from the condition or disease, those susceptible to the condition or disease, or those intending to prevent the condition or disease.

[0032] In this invention, the target of prevention and control is a human or mammal, including bovines, equines, sheep, pigs, canines, felines, rodents, and primates.

[0033] The traditional Chinese medicine composition of this invention includes Sedum sarmentosum, Sargassum, Oyster shell, Crataegus pinnatifida, Siegesbeckia orientalis, Schisandra chinensis, and Polygonum cuspidatum. This invention has found that this composition can significantly reduce urinary protein levels, regulate serum ALB, Cr, and BUN levels, improve renal function, and has a good therapeutic effect on nephrotic syndrome. Moreover, this traditional Chinese medicine composition is an existing prepared Chinese medicine, safe to use, and its preparation method is simple, rapid, efficient, and easy to operate. The prepared mixture, tablets, capsules, and micro-pellets are easy to carry and convenient to take. Detailed Implementation

[0034] This invention provides the application of traditional Chinese medicine compositions in the preparation of drugs for the prevention and treatment of nephrotic syndrome. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0035] This invention proposes the application of a traditional Chinese medicine composition in the preparation of a drug for treating nephrotic syndrome. The composition comprises: 1-1000 parts of *Sedum sarmentosum*, 1-500 parts of *Sargassum fusiforme*, 1-1000 parts of oyster shell, 1-500 parts of hawthorn, 1-1000 parts of *Siegesbeckia orientalis*, 1-500 parts of *Schisandra chinensis*, and 1-800 parts of *Polygonum cuspidatum*. The application can be any method beneficial to improving the corresponding symptoms of the patient, including treatment or prevention. The composition of this invention can be directly ground into powder or can be an extract obtained by conventional methods in the art. The traditional Chinese medicines used in the composition of this invention can also be used by directly grinding into powder, extracting, or other processed forms.

[0036] Furthermore, the traditional Chinese medicine composition includes: 1-1000 parts of Sedum sarmentosum, 1-500 parts of Sargassum, 1-1000 parts of Oyster shell, 1-500 parts of Crataegus pinnatifida, 1-1000 parts of Siegesbeckia orientalis, 1-500 parts of Schisandra chinensis, and 1-800 parts of Polygonum cuspidatum.

[0037] Further, the traditional Chinese medicine composition comprises: 500-1000 parts of Sedum sarmentosum, 200-400 parts of Sargassum, 500-1000 parts of Ostrea gigas, 200-400 parts of Crataegus pinnatifida, 500-1000 parts of Siegesbeckia orientalis, 200-400 parts of Schisandra chinensis, and 300-700 parts of Polygonum cuspidatum.

[0038] Preferably, the traditional Chinese medicine composition comprises: 700-950 parts of Sedum sarmentosum, 200-350 parts of Sargassum, 700-950 parts of Ostrea gigas, 200-350 parts of Crataegus pinnatifida, 700-950 parts of Siegesbeckia orientalis, 200-350 parts of Schisandra chinensis, and 500-650 parts of Polygonum cuspidatum.

[0039] Most preferably, the traditional Chinese medicine composition includes: 923 parts of Siegesbeckia orientalis, 923 parts of Sedum sarmentosum, 308 parts of Sargassum, 923 parts of Ostrea gigas, 308 parts of Crataegus pinnatifida, 618 parts of Polygonum cuspidatum, and 308 parts of Schisandra chinensis.

[0040] Specifically, the medication for treating nephrotic syndrome is selected from oral dosage forms, injectable dosage forms, or topical dosage forms.

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

[0042] The composition of the present invention described above can be made by directly grinding the ingredients into powder using traditional methods, or by obtaining extracts through conventional means, so as to prepare different dosage forms, achieve better therapeutic effects, and be more conducive to the preparation of modern pharmaceutical dosage forms.

[0043] The present invention also proposes a method for preparing any of the aforementioned traditional Chinese medicine compositions, wherein the preparation method is as follows:

[0044] Siegesbeckia orientalis, Sedum sarmentosum, seaweed, calcined oyster shell, hawthorn, Polygonum cuspidatum, and Schisandra chinensis were extracted by reflux with water, adding 8 times the amount of water each time, and extracting for 1.5 hours. The extract was filtered, and the filtrate was concentrated under reduced pressure at no higher than 80℃ to obtain an extract with a relative density of 1.25-1.30 (60±5℃). The extract was then dried under vacuum using a belt dryer, pulverized, sieved, and mixed with sucralose and dextrin to obtain granules.

[0045] The compound combination drug for treating nephrotic syndrome of this invention can significantly reduce urinary protein levels in rats with nephrotic syndrome, increase serum ALB levels, and decrease Cr and BUN levels, thereby improving renal function in rats with nephrotic syndrome. Furthermore, the preparation method of the compound combination drug for treating nephrotic syndrome is simple, rapid, efficient, and easy to operate. The prepared mixtures, tablets, capsules, and micro-pellets are easy to carry and convenient to take.

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

[0047] Unless otherwise specified, all experiments in the following experiments were conducted under standard conditions or conditions recommended by the manufacturer. Active pharmaceutical ingredients (APIs) or excipients, as well as reagents or instruments whose manufacturers are not specified, are all commercially available products. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight.

[0048] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. The invention is further illustrated below with reference to embodiments:

[0049] Example 1: Preparation of the Traditional Chinese Medicine Composition

[0050] Seven medicinal herbs, including 923g of Siegesbeckia orientalis, 923g of Sedum sarmentosum, 308g of Sargassum, 923g of Ostrea gigas, 308g of Crataegus pinnatifida, 618g of Polygonum cuspidatum, and 308g of Schisandra chinensis, were extracted twice by reflux with water, adding 8 times the amount of water each time, for 1.5 hours each time. The extract was filtered, and the filtrate was concentrated under reduced pressure at no higher than 80℃ to obtain an extract with a relative density of 1.25-1.30 (60±5℃). The extract was then dried under vacuum using a belt dryer, pulverized, and sieved to obtain an intermediate of the composition. 2g of sucralose and an appropriate amount of dextrin were added, mixed well, and 1000g of granules were obtained.

[0051] Experimental Example 2: The therapeutic effect of a traditional Chinese medicine composition on doxorubicin-induced nephrotic syndrome in rats

[0052] 1. Experimental Materials

[0053] 1.1 Animals

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

[0055] Breeding environment: room temperature 20-26℃, relative humidity controlled at 40%-70%, animal stocking density 5 animals / cage.

[0056] 1.2 Drugs

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

[0058] 1.3 Instruments

[0059] Refrigerator (Haier, model: BCD-216SZ); Electronic balance (Sartorius Scientific Instruments, model: BSA224S-CW); Purified water instrument (Milli-QPlus, Germany); Animal weight scale (Shanghai Xiangchuan Electronic Weighing Instrument Co., Ltd., model: CX-SC-DA); Multifunctional microplate reader (MD, USA, model: Flexstation 3).

[0060] 2. Dosage design

[0061] The daily dosage of the traditional Chinese medicine composition is 8.62g of crude drug / day. Based on the body surface area calculation method, the equivalent dose for rats is 8.62g / 60kg × 6.2 = 0.89g of crude drug / kg. Following a 1 / 4:1 / 2:1 dosing regimen, the low dose is 0.22g of crude drug / kg, the medium dose is 0.45g of crude drug, and the high dose is 0.89g. Administered by gavage, 0.1mL / 10g, once daily.

[0062] The daily dose of benazepril tablets is 10 mg / day. Based on the body surface area calculation 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 acclimatized for 3 days with free access to feed and water. After normal urine protein levels were detected by urine protein test strips, all rats except the normal control group were injected intravenously with doxorubicin hydrochloride (6 mg / kg) once per dose to establish a rat model of doxorubicin nephropathy syndrome. The success of model establishment was determined by measuring the urine protein content of each rat. Rats with successful model establishment were randomly divided into a model control group, a benazepril tablet group, and high, medium, and low dose groups of the traditional Chinese medicine combination. After grouping, the animals were administered the medicine by gavage once daily for 2 weeks. Dosage for each group: benazepril tablet group (1.1 mg / kg), low, medium, and high dose groups of the traditional Chinese medicine combination (0.22 g / kg, 0.45 g / kg, and 0.89 g / kg, respectively).

[0065] During the two-week drug administration cycle, rats in each group were placed in clean metabolic cages once a week to collect 24-hour urine. The 24-hour urinary protein excretion was measured using the biuret method. After the experiment, 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℃ for later analysis.

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

[0067] 4. Statistical Methods

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

[0069] 5. Experimental Results

[0070] 5.1 Effects on urinary protein levels in rats with doxorubicin-induced nephrotic syndrome

[0071] Compared with the normal control group, the urinary protein levels in the model group rats were significantly increased in the first and second weeks after drug administration (P<0.001), indicating successful model establishment. Compared with the model group, the urinary protein levels in the benazepril tablet group were significantly decreased (P<0.001), and the urinary protein levels in the medium and high dose groups of the traditional Chinese medicine composition were also significantly decreased (P<0.01), indicating that the traditional Chinese medicine composition has a good ameliorative effect on doxorubicin-induced kidney damage in rats. The results are shown in Table 1.

[0072] Table 1. Effects of doxorubicin-induced nephrotic syndrome on urinary protein levels in rats.

[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 doxorubicin-induced nephrotic syndrome

[0076] Compared with the normal control group, the serum ALB, Cr, and BUN levels in the model group rats were significantly increased (P<0.01). After treatment with benazepril tablets, compared with the model control group, the serum ALB level in the benazepril tablet group was significantly increased (P<0.01), while the Cr and BUN levels were significantly decreased (P<0.01). Treatment with the traditional Chinese medicine combination at medium and high doses also significantly increased the serum ALB level and decreased the Cr and BUN levels in rats (P<0.05, P<0.01). The results are shown in Table 2.

[0077] Table 2 Effects of doxorubicin-induced nephrotic syndrome on serum biochemical parameters 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 Conclusions

[0081] The traditional Chinese medicine composition at concentrations of 0.45 and 0.89 g crude drug / kg significantly regulated the urinary protein content and serum ALB, Cr, and BUN biochemical indicators in rats with doxorubicin-induced nephrotic syndrome, indicating that the traditional Chinese medicine composition has a certain ameliorative effect on renal function in rats with doxorubicin-induced nephrotic syndrome.

[0082] Experimental Example 3: The therapeutic effect of a traditional Chinese medicine composition on a rat model of nephrotic syndrome induced by puromycin aminonucleoside

[0083] 1. Experimental Materials

[0084] 1.1 Animals

[0085] Sixty male SD rats, SPF grade, weighing 250–270 g, were purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0086] Rearing environment: room temperature 20-26℃, relative humidity controlled at 40%-70%, and stocking density of 5 birds / cage.

[0087] 1.2 Drugs

[0088] The traditional Chinese medicine composition was prepared according to the method in Example 1 and was provided by Jiangsu Kangyuan Pharmaceutical Co., Ltd.; benazepril tablets (No. 2007, Novartis Pharmaceuticals, Beijing); 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 Biotechnology, catalog number: RF10966); rat ALB ELISA kit (Nanjing Jiancheng Bioengineering Institute, catalog 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); physiological 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 system (Milli-Q Plus, Germany); Animal weight scale (Shanghai Xiangchuan Electronic Weighing Instrument Co., Ltd., model CX-SC-DA); Microplate reader (McGen, USA, model Flexstation3).

[0091] 2. Dosage design

[0092] The daily dosage of the traditional Chinese medicine composition is 8.62g of crude drug / day. Based on the body surface area calculation method, the equivalent dose for rats is 8.62g / 60kg × 6.2 = 0.89g of crude drug / kg. Following a 1 / 4:1 / 2:1 dosing regimen, the low dose is 0.22g of crude drug / kg, the medium dose is 0.45g / kg, and the high dose is 0.89g / kg. Administered by gavage, 0.1mL / 10g, once daily.

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

[0094] 3. Experimental Methods

[0095] Sixty Wistar rats were acclimatized for 3 days with free access to food and water. After normal urine protein levels were detected by urine protein test strips, they were randomly divided into 6 groups using a random number table: normal control group, model control group, dexamethasone control group, and high, medium, and low dose groups of the traditional Chinese medicine composition, with 10 rats in each group. After successful anesthesia with intraperitoneal injection of 10% chloral hydrate (0.33 g / kg body weight), the rats were fixed in a supine position, their necks were prepared, and disinfected with 75% alcohol. Except for the normal control group, rats in all other groups underwent jugular vein cannulation and slow injection of PAN saline solution at 40 mg / kg body weight. The control group underwent jugular vein cannulation and slow injection of the same volume of 0.9% saline. On the 5th day after jugular vein cannulation, all rats were placed in clean metabolic cages, fasted, and given water. 24-hour urine was collected, and 24-hour urinary protein excretion was measured using the biuret method. A significant increase in urinary protein compared to the control group was considered a successful model establishment criterion. Meanwhile, starting from day 6 of the experiment, each group was given intraperitoneal injection and oral gavage, respectively. The benazepril tablet group (1.1 mg / kg) and the traditional Chinese medicine combination at low, medium and high doses (0.22 g / kg, 0.45 g / kg and 0.89 g / kg) were given once a day for two consecutive weeks.

[0096] During the two-week drug administration cycle, rats in each group were placed in clean metabolic cages once a week to collect 24-hour urine. The 24-hour urinary protein excretion was measured using the biuret method. After the experiment, rats were anesthetized by intraperitoneal injection of 10% chloral hydrate, and blood was collected by puncture of the abdominal aorta using a syringe. After standing at room temperature for 3 hours, the serum was separated and stored at -20℃ for later analysis.

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

[0098] 4. Statistical Methods

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

[0100] 5. Experimental Results

[0101] 5.1 Effects on urinary protein levels in rats with puromycin aminonucleoside-induced nephrotic syndrome

[0102] Compared with the normal control group, the urinary protein levels in the model group rats were significantly increased in the first and second weeks of drug administration (P<0.001); compared with the model group, the urinary protein levels in the benazepril tablet group were significantly decreased (P<0.001), and the urinary protein levels in the medium and high dose groups of the traditional Chinese medicine composition were significantly decreased (P<0.01). The results are shown in Table 3.

[0103] Table 3 Effects of puromycin aminonucleoside-induced nephrotic syndrome on urinary protein levels in rats

[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 of Puromycin Aminonucleotide-Induced Nephrotic Syndrome on Serum Biochemical Indicators in Rats: Compared with the normal control group, the serum ALB, Cr, and BUN levels in the model group rats were significantly increased (P<0.001). After treatment with benazepril tablets, compared with the model control group, the serum ALB, Cr, and BUN levels in the benazepril tablet group were significantly decreased (P<0.001). The medium and high doses of the traditional Chinese medicine combination significantly increased the serum ALB content in rats (P<0.01, P<0.001) and significantly decreased the Cr and BUN content (P<0.01, P<0.001). The results are shown in Table 4.

[0107] Table 4. Effects of serum biochemical parameters on puromycin-aminonucleoside-induced nephrotic syndrome in rats

[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 Conclusions

[0111] The traditional Chinese medicine composition at concentrations of 0.45 and 0.89 g crude drug / kg significantly reduced urinary protein levels in rats with puromycin aminonucleoside-induced nephrotic syndrome, and significantly increased serum ALB levels and decreased Cr and BUN levels, indicating that the traditional Chinese medicine composition has a relieving and improving effect on kidney damage in rats with nephrotic syndrome, and that the traditional Chinese medicine composition has a certain therapeutic effect on the rat model of puromycin aminonucleoside-induced nephrotic syndrome.

[0112] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The application of a traditional Chinese medicine composition in the preparation of a drug for treating nephrotic syndrome, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 700-950 parts of Sedum sarmentosum, 200-350 parts of Sargassum, 700-950 parts of Oyster shell, 200-350 parts of Crataegus pinnatifida, 700-950 parts of Siegesbeckia orientalis, 200-350 parts of Schisandra chinensis, and 500-650 parts of Polygonum cuspidatum.

2. The application according to claim 1, characterized in that, The nephrotic syndrome mentioned is drug-induced nephrotic syndrome.

3. The application according to claim 2, characterized in that, The drug is doxorubicin and / or puromycin aminonucleoside.

4. The application according to any one of claims 1 to 3, characterized in that, The preparation method of the traditional Chinese medicine composition includes: adding water to extract the following herbs: Siegesbeckia orientalis, Sedum sarmentosum, Sargassum, Ostrea gigas, Crataegus pinnatifida, Polygonum cuspidatum, and Schisandra chinensis, adding 5-10 times the weight of water each time, extracting for 2-4 hours, filtering, concentrating the filtrate under reduced pressure at no higher than 80℃ to an extract with a relative density of 1.25-1.30 at 60±5℃, and then concentrating and drying.

5. The application according to claim 4, characterized in that, In the reflux extraction, reflux extraction is performed twice, each time for 1.5 hours.

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