A traditional Chinese medicine composition for treating IgA nephropathy, a preparation method and application thereof

Abstract

The application relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for treating IgA nephropathy, which is prepared from the following raw medicinal materials in parts by weight: radix codonopsis 9-15 parts, radix astragali 9-15 parts, zingiberis rhizoma 6-9 parts, rhizoma pinelliae praeparatum 9-15 parts, radix scrophulariae 6-9 parts, rhizoma coptidis 3-6 parts, coix lacryma-jobi 15-45 parts, pyrrosia 9-15 parts, euonymus alatus 9-15 parts and licorice 3-9 parts. The application also includes the application of the traditional Chinese medicine composition in the preparation of medicines for treating IgA nephropathy. The application ingeniously applies the modern intractable disease to the formula, embodies the treatment principle of treating different diseases in the same way, the selected raw medicinal materials meet the compatibility principle of monarch, minister, assistant and messenger, have the effect of treating the principal and secondary symptoms at the same time, and the application effect is remarkable. The application provides a new treatment scheme for patients with IgA nephropathy, improves the survival rate of the patients, and has a good application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine, in particular to a traditional Chinese medicine composition for treating IgA nephropathy, a preparation method and application thereof. BACKGROUND

[0002] IgA nephropathy (IgAN) refers to a primary glomerular disease in which IgA immune complexes are deposited in the mesangial area of the glomerulus, accompanied by mesangial cell proliferation and increased matrix. IgAN is the most common primary glomerular nephritis worldwide. It is the main cause of chronic kidney disease, and up to 40% of patients develop end-stage renal disease about 20 years after diagnosis. IgAN has a very diverse geographic distribution, and is more common in Asian populations and young adults.

[0003] The widely accepted mechanism of IgAN is the multiple hit theory. Abnormal glycosylation of IgA1 under the influence of multiple factors such as environment or genes is the first step in the pathogenesis of IgAN, so tracing the generation of Gd-IgA1 is particularly important. The "gut-kidney axis" theory of IgAN was first proposed by Coppo, who believed that immune tolerance defects could lead to abnormal reactions of the microbiota and changes in the intestinal barrier, including increased absorption of digestive tract antigens and bacterial toxins, triggering mucosa-associated lymphoid tissue activation and subclinical intestinal inflammation. This can produce abnormal reactions to digestive tract antigens or commensal microorganisms, synthesize Gd-IgA1, and initiate the multiple hits of IgAN. Genome-wide association studies have also confirmed that most of the loci associated with IgAN risk are related to immune-mediated inflammatory bowel disease, maintenance of the intestinal barrier, and regulation of intestinal flora responses, and it can be inferred that genetic and environmental factors may interact to induce functional abnormalities in intestinal mucosal lymphoid tissue, leading to the development of IgAN. More and more studies have confirmed that dysfunction of the intestinal mucosal barrier is involved in the occurrence and development of IgAN, and is the basis and source of IgAN.

[0004] Chinese patent document CN106310146B discloses a traditional Chinese medicine composition for preventing and treating IgA nephropathy and its preparation method, which is made from 15-25 parts of Rehmannia glutinosa, 25-35 parts of Fagopyrum dibotrys, 15-25 parts of Ligustrum lucidum, 5-15 parts of Cicadae periostracum, 5-15 parts of Platycodon grandiflorus, 15-25 parts of Scrophularia ningpoensis, 5-15 parts of Oroxylum indicum, 5-15 parts of Terminalia chebula, 10-20 parts of Dioscorea nipponica, 10-20 parts of Typha orientalis, 5-15 parts of Curcuma longa, 15-25 parts of Forsythia suspensa, 10-20 parts of Euonymus alatus, 15-25 parts of Ophiopogon japonicus, and 5-15 parts of Bombyx mori. Chinese patent CN108714205A discloses a traditional Chinese medicine granule for treating IgA nephropathy of spleen and kidney yang deficiency type, its preparation method, and its application, which is made from 9 parts of Astragalus membranaceus, 9 parts of Cinnamomum cassia, 9 parts of Paeonia lactiflora, 18 parts of Zingiber officinale, and 12 parts of Ziziphus jujuba. Chinese patent document CN105920453A discloses a traditional Chinese medicine composition for treating IgA nephropathy of spleen and kidney qi deficiency type, which is made from 25 parts of Rosa laevigata root, 15 parts of Psoralea corylifolia, 20 parts of Aralia elata, 9 parts of Rhizoma Sedum, 15 parts of Rhizoma Cyathulae, 10 parts of Rhizoma Sedum, 20 parts of Rhizoma Lysimachiae, 15 parts of Herba Sedum, 10 parts of Herba Lysimachiae, 15 parts of Herba Sedum, 10 parts of Herba Prunellae, 15 parts of Herba Achyranthis Bidentatae, 8 parts of Radix Salviae Miltiorrhizae, 10 parts of Herba Ziziphi Spinosae, 8 parts of Herba Cyperi, 20 parts of Herba Lysimachiae, and 8 parts of Radix Glycyrrhizae. Chinese patent document CN105816681A discloses a traditional Chinese medicine composition for treating IgA nephropathy of Qi and Yin deficiency type, which is made from 10 parts of Acanthopanax senticosus, 20 parts of Cyperus rotundus, 15 parts of Caragana korshinskii root, 15 parts of Deer Herb, 15 parts of Smilax china, 10 parts of Clematis armandii, 20 parts of Artemisia argyi branch, 10 parts of Achyranthes bidentata, 20 parts of Napier grass, 15 parts of Smilax china, 20 parts of Codonopsis pilosula, 25 parts of Euonymus fortunei, 9 parts of Lysimachia christinae, 20 parts of Gynostemma pentaphyllum, and 8 parts of Glycyrrhiza uralensis. However, no reports have been found regarding a traditional Chinese medicine composition for treating IgA nephropathy, its preparation method, and its application. Summary of the Invention

[0005] The purpose of this invention is to provide a traditional Chinese medicine composition for treating IgA nephropathy, its preparation method, and its application.

[0006] Licorice Decoction for Clearing the Heart is a classic formula in Traditional Chinese Medicine. Modern mechanistic studies have shown that it has anti-inflammatory and analgesic effects, as well as free radical scavenging effects. By clearing free radicals, it prevents oxidative damage to the mucosa, inhibits inflammatory responses in the mucosa, protects the gastric mucosa, promotes ulcer healing, protects the liver, enhances immunity, and improves the body's resistance to hypoxia. Based on previous clinical experience, this invention combines Licorice Decoction for Clearing the Heart with Xiaobai Decoction, creating a modified version of the formula. This formula uses both warming and cooling herbs to harmonize Yin and Yang, combines bitter and pungent herbs to restore the ascending and descending functions, and combines tonification and purgation to regulate deficiency and excess, thus treating both the root cause and the symptoms. IgAN presents with a mixture of deficiency and excess, and a complex interplay of warming and cooling symptoms. Since the symptoms are similar, the treatment should also be similar. The treatment method should be based on the symptoms, the formula should be derived from the treatment method, and the formula should be formulated based on the herbs, reflecting the TCM principle of "treating different diseases with the same method."

[0007] The first aspect of the present application provides a traditional Chinese medicine composition for treating IgA nephropathy, which is prepared from raw medicinal materials in the following proportions by weight: Radix Codonopsis 9-15 parts, Radix Astragali 9-15 parts, Rhizoma Zingiberis 6-9 parts, Rhizoma Pinelliae 9-15 parts, Radix Scrophulariae 6-9 parts, Rhizoma Coptidis 3-6 parts, Radix Alpiniae 15-45 parts, Radix Pyrrosiae 9-15 parts, Radix Taxilli 9-15 parts, and Radix Glycyrrhizae Preparata 3-9 parts.

[0008] Further, the traditional Chinese medicine composition for treating IgA nephropathy is prepared from raw medicinal materials in the following proportions by weight: Radix Codonopsis 12-15 parts, Radix Astragali 12-15 parts, Rhizoma Zingiberis 8-9 parts, Rhizoma Pinelliae 12-15 parts, Radix Scrophulariae 8-9 parts, Rhizoma Coptidis 5-6 parts, Radix Alpiniae 30-45 parts, Radix Pyrrosiae 12-15 parts, Radix Taxilli 12-15 parts, and Radix Glycyrrhizae Preparata 6-9 parts.

[0009] Still further, the traditional Chinese medicine composition for treating IgA nephropathy is prepared from raw medicinal materials in the following proportions by weight: Radix Codonopsis 15 parts, Radix Astragali 15 parts, Rhizoma Zingiberis 9 parts, Rhizoma Pinelliae 15 parts, Radix Scrophulariae 9 parts, Rhizoma Coptidis 6 parts, Radix Alpiniae 30 parts, Radix Pyrrosiae 15 parts, Radix Taxilli 15 parts, and Radix Glycyrrhizae Preparata 9 parts.

[0010] The second aspect of the present application provides use of the traditional Chinese medicine composition for treating IgA nephropathy as described above in the preparation of a medicament for treating IgA nephropathy.

[0011] Further, the IgA nephropathy includes chronic glomerulonephritis.

[0012] The third aspect of the present application provides a pharmaceutical preparation taking the traditional Chinese medicine composition for treating IgA nephropathy as described above as an active ingredient, which is prepared into a clinically acceptable conventional pharmaceutical preparation according to a conventional preparation method of traditional Chinese medicine.

[0013] Further, the pharmaceutical preparation is in the form of a decoction, granules, powder, capsule, tablet, mixture, or oral liquid.

[0014] Further, the pharmaceutical preparation further comprises a pharmaceutically acceptable carrier or excipient.

[0015] Further, the active ingredient of the pharmaceutical preparation is prepared by water extraction or extraction with 40-80% (volume percentage) ethanol.

[0016] Still further, the preparation method of the pharmaceutical preparation comprises the following steps:

[0017] The raw medicinal materials are weighed and added with water for extraction for 2-3 times, with the amount of water added corresponding to 6-12 times the total weight of the medicinal materials each time, and the extraction time being 1-3 hours each time. The extraction liquids are combined, filtered, and concentrated to a clear extract with a relative density of 1.10-1.25 at 70-80°C, thereby obtaining the active ingredient of the medicament.

[0018] Further, the preparation method of the traditional Chinese medicine composition for treating IgA nephropathy comprises the following steps:

[0019] According to the weight ratio, each raw material is taken, and water is added for extraction 2-3 times, the water amount is 6-12 times of the total weight of the medicinal materials each time, and the extraction time is 1-3 hours each time, the extraction liquid is combined, filtered, and concentrated to a clear extract with a relative density of 1.05-1.20 at 70-80 DEG C, ethanol is added to make the alcohol content 40-70%, and then standing for 12-24 hours, filtering, and concentrating the filtrate to a relative density of 1.10-1.25 at 70-80 DEG C, so that the active ingredient of the medicine is obtained.

[0020] In a fourth aspect, the application provides use of the pharmaceutical preparation as described above in the preparation of a medicament for treating IgA nephropathy.

[0021] The application provides the following solutions:

[0022] The original prescription of Gancao Xiexin decoction is composed of Radix Glycyrrhizae (charred) 4 liang, Radix Scrophulariae 3 liang, Rhizoma Zingiberis Recens 3 liang, Rhizoma Pinelliae (washed) half a tael, Fructus Jujubae (crushed) 12 pieces, Rhizoma Coptidis 1 liang, and Radix Ginseng 3 liang. IgAN is a syndrome of deficiency and excess, and excessive supplementation can easily close the door and leave the enemy, so the Radix Ginseng and Fructus Jujubae are removed, the charred Radix Glycyrrhizae is reduced, the Radix Paeoniae Alba is replaced by the Radix Paeoniae Alba with the property of being sweet and warm, and the Radix Astragali is combined, both of which are the monarchs, and the function is to warm the middle and dispel cold, tonify qi and invigorate the spleen, so as to relieve the symptoms of diarrhea, edema and other symptoms caused by the deficiency of the spleen and stomach, and the failure of water and food to be digested. The Radix Astragali can also consolidate the kidney and astringe essence, reduce the loss of subtle substances, and play the role of "blocking flow". Rhizoma Pinelliae and Rhizoma Zingiberis Recens are both pungent and warm, and enter the stomach meridian, and the intention is to resolve phlegm and eliminate stagnation, descend adverse flow and stop vomiting, so that the stomach qi is properly descended. In this way, the physiological functions of the spleen and stomach can be exerted, so that water and food are properly transported, the qi movement is properly promoted, and yin and yang and dampness are properly coordinated. Spleen deficiency leads to kidney deficiency, and qi deficiency develops into yang deficiency, and the dampness is a yin evil, and tends to descend, so the body is characterized by deficiency of spleen and kidney yang and cold dampness, and due to the abnormality of the qi movement of the spleen and stomach, the middle jiao is not unobstructed, yin is excessive in the lower, and the deficiency of yang is floating upward, so there are manifestations of upper heat and lower cold, in addition to the manifestations of lower cold deficiency such as edema of lower limbs, loose stool, cold and soreness of waist and knees, there are also manifestations of heat such as sore throat and mouth sores, and if the dampness is converted into damp-heat evil for a long time, the Rhizoma Coptidis and Rhizoma Coptidis are used to treat it, both of which are bitter and cold, and the bitter and cold descending and discharging can not only eliminate the damp-heat in the middle jiao, but also descend the floating deficiency heat, and also can regulate the heat of other drugs. The Radix Alpiniae Oxophylla and Radix Pyrrosiae can clear damp-heat and benefit water-dampness, and both of them are the auxiliary drugs, so that the damp evil is excreted with urine, and the effect of resolving edema is strengthened; the Arisaema amurense Maxim. can break blood and disperse stasis, and "blood not excreted is water", so that the collaterals are unobstructed and water is excreted.

[0023] The application has the following advantages:

[0024] 1. This invention ingeniously applies classical Chinese medicine formulas to modern intractable diseases, embodying the principle of "treating different diseases with the same method." The ingredients in the formula conform to the principle of "principal, assistant, adjuvant, and guide" in their compatibility. The formula uses both warming and cooling herbs to harmonize Yin and Yang, combines bitter and pungent herbs to restore the ascending and descending functions, and employs both tonifying and purging methods to regulate deficiency and excess. It has the effect of treating both the root cause and the symptoms, resulting in significant therapeutic effects. The ingredients work synergistically, leading to excellent efficacy; furthermore, the optimal ratio of the ingredients has been determined through experimental screening, resulting in significant therapeutic benefits.

[0025] 2. The active pharmaceutical ingredients of this invention are widely available and the drug preparation method is simple.

[0026] 3. Based on the gut-kidney axis theory, this invention provides a new treatment option for patients with IgA nephropathy, improves the survival rate of such patients, and has a promising application prospect. Attached Figure Description

[0027] Figure 1 IgA immunofluorescence in rats of each group (400×).

[0028] Figure 2 HE staining of rat kidneys in each group (200×).

[0029] Figure 3 Masson staining of rat kidneys (200×) in each group.

[0030] Figure 4 PAS staining (200×) for kidney pathology in rats of each group.

[0031] Figure 5 HE staining of intestinal tissue from rats in each group (200×).

[0032] Figure 6 Expression of TLR4, MyD88, NF-κB, IL-6, TNF-α, and MCP-1 proteins in the kidney tissues of rats in each group; compared with the normal group, ** P < 0.01; * P < 0.05; compared with the model group, # P < 0.05 ## P < 0.01.

[0033] Figure 7 Expression of Claudin-1, Occludin, and ZO-1 proteins in the intestinal tissue of rats in each group; compared with the normal group, ** P < 0.01; * P < 0.05; compared with the model group, # P < 0.05 ## P < 0.01.

[0034] Figure 8The protein expression of TLR4, MyD88, NF-κB, IL-6, TNF-α and MCP-1 in the intestinal tissue of the rats in each group was detected, and the results were compared with those in the normal group, ** P<0.01; * P<0.05; compared with the model group, # P<0.05, ## P<0.01. DETAILED DESCRIPTION

[0035] The specific embodiment of the present application is described in detail below in combination with examples.

[0036] Example 1: the traditional Chinese medicine composition (I) of the present application

[0037] Dangshen 9 parts, Huangqi 9 parts, Ganjiang 6 parts, Shihai 9 parts, Huanglian 3 parts, Yiyiren 15 parts, Shiwve 9 parts, Guijian 9 parts, Zhigancao 3 parts.

[0038] Example 2: the traditional Chinese medicine composition (II) of the present application

[0039] Dangshen 10 parts, Huangqi 10 parts, Ganjiang 7 parts, Shihai 10 parts, Huanglian 4 parts, Yiyiren 20 parts, Shiwve 10 parts, Guijian 10 parts, Zhigancao 4 parts.

[0040] Example 3: the traditional Chinese medicine composition (III) of the present application

[0041] Dangshen 12 parts, Huangqi 12 parts, Ganjiang 8 parts, Shihai 12 parts, Huanglian 5 parts, Yiyiren 40 parts, Shiwve 12 parts, Guijian 12 parts, Zhigancao 6 parts.

[0042] Example 4: the traditional Chinese medicine composition (IV) of the present application

[0043] Dangshen 15 parts, Huangqi 15 parts, Ganjiang 9 parts, Shihai 15 parts, Huanglian 6 parts, Yiyiren 45 parts, Shiwve 15 parts, Guijian 15 parts, Zhigancao 8 parts.

[0044] Example 5: the traditional Chinese medicine composition (V) of the present application

[0045] Dangshen 15 parts, Huangqi 15 parts, Ganjiang 9 parts, Shihai 15 parts, Huanglian 6 parts, Yiyiren 30 parts, Shiwve 15 parts, Guijian 15 parts, Zhigancao 9 parts.

[0046] Example 6: the preparation method (I) of the traditional Chinese medicine composition of the present application

[0047] The method comprises the following steps: taking each raw medicinal material according to the weight ratio of any one of examples 1-5, extracting 2-3 times with water, the amount of water added each time being 6-12 times the total weight of the medicinal materials, and the extraction time being 1-3 hours each time, combining the extraction liquid, filtering, and concentrating the filtrate to a clear extract with a relative density of 1.10-1.25 at 70-80°C, thereby obtaining the medicinal active ingredient.

[0048] Example 7: Preparation method of the traditional Chinese medicine composition of the application (two)

[0049] The method comprises the following steps: taking each raw medicinal material according to the weight ratio of any one of examples 1-5, extracting 2-3 times with water, the amount of water added each time being 6-12 times the total weight of the medicinal materials, and the extraction time being 1-3 hours each time, combining the extraction liquid, filtering, and concentrating the filtrate to a clear extract with a relative density of 1.10-1.25 at 70-80°C, thereby obtaining the medicinal active ingredient.

[0050] Example 8: Preparation of a decoction

[0051] The raw medicinal materials in the proportions described in examples 1-5 are taken respectively, and a conventional method for preparing a traditional Chinese medicine decoction is used to decoct the raw medicinal materials into a decoction.

[0052] Example 9: Preparation of tablets / capsules

[0053] The medicinal active ingredient obtained in example 6 or 7 is taken, pharmaceutical excipients are added, and a conventional pharmaceutical method is used to compress the mixture into tablets or fill the mixture into capsules.

[0054] Example 10: Preparation of granules

[0055] The medicinal active ingredient obtained in example 6 or 7 is taken, pharmaceutical excipients are added, and a conventional pharmaceutical method is used to prepare granules.

[0056] Example 11: Preparation of a mixture

[0057] The medicinal active ingredient obtained in example 6 or 7 is taken, pharmaceutical excipients are added, and a conventional pharmaceutical method is used to prepare a mixture.

[0058] Example 12: Preparation of an oral liquid

[0059] The medicinal active ingredient obtained in example 6 or 7 is taken, dissolved, purified, and concentrated, conventional oral liquid excipients are added, and a conventional pharmaceutical method is used to prepare the oral liquid.

[0060] Example 13: Animal experiments

[0061] I. Experimental materials

[0062] 1. Animals: 48 SPF male SD rats were purchased from Shanghai Slek Experimental Animal Co., Ltd., with a body weight of 160-180 g, animal qualification certificate number: SYXK (Shanghai) 2020-0009, animal experiment ethics number: PZSHUTCM220822023. The animals were raised in the 4th floor No. 4 feeding room of the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine, with a relative temperature of 20-25℃, a relative humidity of 53-57%, a 12 / 12 hour light-dark alternating environment, and free access to food and water. The animals were adaptively fed for one week.

[0063] 2. Drug composition: The licorice heart-purging derivative formula of the present application is composed of Huangqi 15g, Dangshen 15g, Ganjiang 9g, Zhishanxia 15g, Huangqin 9g, Huanglian 6g, Yiyiren 30g, Shiwu 15g, Guijian 15g, and Zhigancao 9g. It was prepared by the Pharmacy Department of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine. According to the body weight of an adult of 70Kg and the body weight of a rat of 500g, the daily treatment dose of a rat was 12.42g / kg, and the licorice heart-purging derivative formula extract was decocted to a concentration of 3.105g / ml. According to the adult daily dosage of 100mg, the daily treatment dose of a rat was 9mg / kg, and a suspension with a concentration of 2.25mg / ml was prepared with distilled water.

[0064] II. Methods

[0065] 1. Modeling and grouping

[0066] The 48 male SD rats (180-200g) were randomly divided into two groups according to body weight: normal group (12 rats) and model group (36 rats). The IgA nephropathy rat model was established by the BSA+CCl4+LPS scheme, and the specific operation was as follows: BSA, 400mg / kg (4ml / kg), intragastric administration every other day, for 12 weeks; CCl4 was mixed with castor oil at a ratio of 1:3, 0.4ml / rat, subcutaneous injection every week, for 12 weeks; LPS was injected into the tail vein at 0.05mg (0.2ml) / rat at 6 and 8 weeks. The normal group was replaced with an equal amount of normal saline, and the operation was the same as the model group. At the end of the 12th week of the experiment, the rat urine was collected. Three rats from the normal group and six rats from the model group were randomly selected, and blood samples and kidney tissues were collected after deep anesthesia. After determining the success of the rat modeling, 30 IgA nephropathy rats were randomly divided into 10 rats in the model group, 10 rats in the traditional Chinese medicine group, and 10 rats in the western medicine group using SPSS 26.0 software, so that there was no statistically significant difference between the groups.

[0067] 2. Intervention method

[0068] Traditional Chinese medicine group: the invention of the liquorice heart derivative party extract intragastrically 2ml. Western medicine group: the losartan potassium suspension intragastrically 2ml. Normal group, model group were given the same amount of normal saline. According to the body weight adjustment drug intragastric volume. Continuous 8 weeks, daily timing of drug administration.

[0069] 3, collection and processing of specimens

[0070] (1) urine specimens: disinfection and assembly of metabolic cage in advance; rats were fasted but not water restricted on the day of urine collection; 3ml of fresh urine was collected, and 24-hour urine was collected and accurately measured.

[0071] (2) blood samples: anesthesia: intraperitoneal injection of 2.5% sodium pentobarbital (0.2ml / 100g) to anesthetize rats; blood collection: open the abdominal cavity after the rats are deeply anesthetized, and collect blood from the abdominal aorta; sacrifice: after blood collection is completed, the abdominal aorta is cut off and the rats are sacrificed.

[0072] (3) fecal tissue: collect rat feces, stick about 0.5g of feces on a cotton swab and place it in 4.5ml phosphate buffered saline (1×PBS) for shaking.

[0073] (4) kidney tissue: after blood collection, remove the left kidney of the rat, and peel off the perirenal capsule; wash the kidney with normal saline, divide the kidney into two parts along the coronal plane, and cut off a 1×1×1cm tissue at the renal portal, which is placed in 4% paraformaldehyde solution for later use; the remaining tissue is divided into 1.5ml cryogenic tubes and placed in a liquid nitrogen tank for later use.

[0074] (5) intestinal tissue: after blood collection, take about 1cm of intestinal tube at the ileocecal junction of the rat, and take about 5cm of ileum; wash the intestinal tube with normal saline, cut the ileocecal junction of the intestinal tube into 1×1cm tissue, and attach it to filter paper, which is placed in 4% paraformaldehyde solution for later use; the remaining ileum tissue is divided into 1.5ml cryogenic tubes and placed in a liquid nitrogen tank for later use.

[0075] 4, detection of indicators and methods

[0076] (1) urine specimens: collect fresh urine from rats with metabolic cages, mix well, and take 1ml in a centrifuge tube, centrifuge at 3000rpm / min for 5min at 4℃, discard the supernatant, and collect the urine sediment, mix well, and take 0.2ml evenly on a clean glass slide, cover with a glass slide, and observe under a 10×40 high power microscope, record the results and calculate the average value. Collect 24-hour urine from rats with metabolic cages, mix well, and take 10ml in a centrifuge tube, centrifuge at 3000rpm / min for 10min at 4℃, take 200μl of supernatant, and send it to the laboratory for detection of 24h UP and UACR with a fully automatic biochemical analyzer.

[0077] (2) Blood samples: blood was placed in a coagulation tube for 2 hours, then centrifuged at 3000 rpm / min for 10 min at 4°C using a table centrifuge, 200 μl of supernatant was taken and sent to the laboratory for detection of Scr, BUN, UA and Alb using a full-automatic biochemical analyzer.

[0078] (3) Fecal tissue: after centrifugation of the fecal sample at 3000 rpm for 10 min, the supernatant was taken and stored at -80°C in a refrigerator for detection. Fecal sIgA and serum LPS were detected using an enzyme-linked immunosorbent assay.

[0079] (4) Kidney tissue: the tissue was used to prepare kidney immunofluorescence pathology, and Western-blot was used to detect the protein expression of kidney TLR4, NF-κB p65, MyD88, IL-6, TNF-α and MCP-1.

[0080] (5) Intestinal tissue: the tissue was used to prepare intestinal immunofluorescence kidney pathology, and Western-blot was used to detect the protein expression of intestinal ZO-1, Claudin-1, Occludin, TLR4, NF-κB p65, MyD88, IL-6, TNF-α and MCP-1.

[0081] III. Experimental results

[0082] 1. Changes in urine indicators of rats in each group

[0083] After intervention, compared with the normal group, the urine microscopic red blood cells, urinary red blood cells, 24h UP, 24h mAlb and UACR of the model group of rats were significantly increased (P<0.01); compared with the model group, the indicators of the licorice heart-derived prescription group of the application were significantly reduced (P<0.01), and the 24h UP, 24h mAlb and UACR of the losartan potassium group were significantly reduced (P<0.05). See Table 1.

[0084] Table 1 Changes in urine microscopic red blood cells, urinary red blood cells, 24h UP, 24h mAlb and UACR of rats in each group

[0085]

[0086] Note: compared with the normal group, **P<0.01; *P<0.05; compared with the model group, #P<0.05, ##P<0.01.

[0087] 2. Changes in blood indicators of rats in each group

[0088] After intervention, compared with the normal group, the Scr and BUN of the model group of rats were significantly increased (P<0.01); compared with the model group, the Scr and BUN of the licorice heart-derived prescription group were significantly reduced (P<0.05). See Table 2.

[0089] Scr, BUN, Alb changes of rats in each group

[0090]

[0091] Note: Compared with the normal group, **P<0.01; *P<0.05; compared with the model group, #P<0.05, ##P<0.01.

[0092] After intervention, compared with the normal group, the model group rats IL-6, TNF-α, MCP-1 increased significantly (P<0.01); compared with the model group, the licorice heart-derived group of each index decreased significantly (P<0.01). See Table 3.

[0093] Table 3 IL-6, TNF-α, MCP-1 changes of rats in each group

[0094]

[0095] Note: Compared with the normal group, **P<0.01; *P<0.05; compared with the model group, #P<0.05, ##P<0.01.

[0096] 3, the changes of intestinal function index of rats in each group

[0097] After intervention, compared with the normal group, the model group rats LPS, fecal sIgA increased significantly (P<0.01); compared with the model group, the licorice heart-derived group of each index decreased significantly (P<0.01). See Table 4.

[0098] Table 4 LPS, fecal sIgA changes of rats in each group

[0099]

[0100] Note: Compared with the normal group, **P<0.01; *P<0.05; compared with the model group, #P<0.05, ##P<0.01.

[0101] 4, the changes of pathological conditions of rats in each group

[0102] (1) IgA immunofluorescence of renal tissue

[0103] The IgA deposition in the glomerulus of the rat kidney tissue was detected by immunofluorescence method: no immune complex IgA deposition was observed in the glomerulus of the normal group; the relatively high intensity of IgA deposition in the form of mass or coarse particles was observed in the mesangial area and / or capillary wall of the model group and each treatment group. The IgA immunofluorescence intensity of the model group and the losartan potassium group was higher than that of the normal group (P<0.05); the IgA immunofluorescence intensity of the glycyrrhiza purgation heart derivative prescription group and the losartan potassium group was lower than that of the model group (P<0.05). See Table 5, Figure 1 .

[0104] Table 5 Change in IgA immunofluorescence intensity of rats in each group

[0105]

[0106] Note: compared with the normal group, **P<0.01; *P<0.05; compared with the model group, #P<0.05, ##P<0.01.

[0107] (2) Kidney pathology

[0108] HE staining: it was observed that the glomerular structure of the normal group was normal, and no obvious cell multiplication was observed; the number of cells in the glomerulus of the model group and each treatment group was obviously increased, and part of the capsule cavity was narrowed due to pressure, and the capsule cavity was poorly open. See Figure 2 .

[0109] Masson staining: no obvious change was observed in the glomerulus of the normal group; part of the glomerulus of the IgAN rats in the model group and each treatment group was observed to have local fibrosis change, and blocky, convex red protein deposition to the kidney capsule cavity appeared in the mesangial area, and cell infiltration accompanied by fibrosis was observed in the renal interstitium. See Figure 3 .

[0110] PAS staining: no obvious mesangial proliferation and obvious matrix multiplication were observed in the glomerulus of the normal group; obvious mesangial cells were observed in the glomerulus of the IgAN rats in the model group and each treatment group, accompanied by mesangial matrix proliferation, and different degrees of compression and narrowing of the glomerular capillary loop were found, and glomerular capsule adhesion was observed in part of the glomerular capsule cavity. See Figure 4 .

[0111] (3) Intestinal pathology

[0112] HE staining: no obvious abnormality was observed in the intestinal epithelial cells, intestinal villus height and intestinal crypt depth of the normal group; the intestinal epithelial cells of the model group and each treatment group were exfoliated, the intestinal villi were shortened and irregular, the number of intestinal villi was reduced and the interval was widened. The damage degree of the intestinal barrier of the glycyrrhiza purgation heart derivative prescription group was relieved compared with the model group. See Figure 5 .

[0113] 4. Protein expression in each group of rats

[0114] (1) Kidney tissue

[0115] After intervention, compared with the normal group, the protein expression of TLR4, MyD88, NF-κB, IL-6, TNF-α, and MCP-1 in the model group increased (P<0.05); compared with the model group, the protein expression of TLR4, MyD88, NF-κB, IL-6, TNF-α, and MCP-1 in each treatment group decreased (P<0.05). See Figure 6 .

[0116] (2) Intestinal tissue

[0117] After intervention, compared with the normal group, the protein expression of Claudin-1, Occludin, and ZO-1 in the model group decreased (P<0.05), and the protein expression of TLR4, NF-κB, MyD88, IL-6, TNF-α, and MCP-1 increased (P<0.05); compared with the model group, the protein expression of Claudin-1, Occludin, and ZO-1 in each treatment group increased (P<0.05), and the protein expression of TLR4, MyD88, NF-κB, IL-6, TNF-α, and MCP-1 decreased (P<0.05). See Figure 7 、 8 .

[0118] Example 14: Clinical trial

[0119] I. Clinical data

[0120] 1. Case source: The cases in this study were sourced from the outpatients and inpatients of the Nephrology Department of Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from September 2022 to September 2023, who met the criteria for IgA nephropathy.

[0121] 2. Inclusion criteria: Patients who met the Western medical diagnosis and TCM syndrome diagnosis of IgA nephropathy and had kidney hematuria as the main clinical manifestation; aged 18-65 years, gender and nationality not limited; 24-hour urine protein quantification was 0.5g-3.0g / 24h; urinary sediment red blood cell count was ≥10 / μl; eGFR was ≥30(ml / min / 1.73m 2 )(CKD1-3 stage); did not receive kidney replacement therapy; blood pressure of patients with hypertension was normal or controlled between 120-130 / 70-80mmHg; signed informed consent form.

[0122] 3. Exclusion criteria: Received glucocorticoids, immunosuppressants, cytotoxic drugs within 3 months; suffered from secondary IgA nephropathy such as allergic purpura nephritis, systemic lupus erythematosus; suffered from other types of kidney diseases such as polycystic kidney, pyelonephritis, drug-induced kidney damage; diagnosed as acute renal failure including chronic renal failure with acute exacerbation; combined with malignant tumor, severe cardiovascular and cerebrovascular disease, severe hematological disease, severe liver disease, severe mental illness, severe trauma and other diseases; pregnant or lactating women; kidney pathology percentage of crescent > 25%; allergic or intolerant to the test drug.

[0123] II. Method

[0124] 1. Sample calculation: The study uses the sample size estimation formula for superiority clinical trial (sample size for two-sample rate comparison):

[0125]

[0126] Let the test level = 0.025 (one-sided), the confidence level 1-β = 0.20, π T , π c are the effective rates of the treatment group and the control group, respectively, and the difference between the two groups is clinically significant. According to the previous clinical observation, the effective rate of the licorice heart purging derivative prescription is set to 85%, and previous studies have shown that the effective rate of conventional clinical treatment is 68%. It is calculated that 50 patients are needed in each group, considering a 10% dropout rate, and the total sample size is 110.

[0127] 2. Treatment method: Treatment group: The licorice heart purging derivative prescription granules of the present application (Radix Astragali 15g, Radix Codonopsis 15g, Rhizoma Zingiberis 9g, Rhizoma Pinelliae 15g, Radix Scutellariae 9g, Rhizoma Coptidis 6g, Radix Alpiniae Officinarum 30g, Radix Pyrrosiae 15g, Radix Taxilli 15g, Radix Glycyrrhizae Preparata 9g), 2 times a day, 2 bags each time, and take with boiling water. Control group: Traditional Chinese medicine decoction-free placebo. 2 times a day, 2 bags each time, and take with boiling water. Both groups take continuously for 24 weeks.

[0128] 3. Observation index: Urinary sediment red blood cell count (URBC / μl), urinary microalbumin / creatinine (UACR), 24h urinary protein quantification (24hUP), serum creatinine (Scr), blood urea nitrogen (BUN), estimated glomerular filtration rate (eGFR), plasma albumin (Alb), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein (MCP-1), endotoxin, and TCM syndrome score.

[0129] III. Test results

[0130] 1. Comparison of urine indicators between the two groups of patients

[0131] After treatment, the URBC of the treatment group and the control group decreased compared with before treatment (P<0.05), and the treatment group was significantly lower than the control group (P<0.05); the UACR and 24hUP of the treatment group and the control group decreased compared with before treatment (P<0.05), and there was no significant difference between the groups (P>0.05). See Tables 5-7.

[0132] Table 5 Comparison of urinary sediment red blood cell count levels between the two groups ( / μl)

[0133]

[0134] Note: Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0135] Table 6 Comparison of UACR levels between the two groups (mg / g)

[0136]

[0137] Note: ** P<0.01; * P<0.05. Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0138] Table 7 Comparison of 24hUP levels between the two groups (g / 24h)

[0139]

[0140] Note: Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0141] 2. Comparison of renal function indicators between the two groups

[0142] After treatment, Scr of treatment group and control group decreased compared with before treatment (P<0.05), and Scr of treatment group was significantly lower than that of control group (P<0.05); BUN of treatment group decreased compared with before treatment (P<0.05), and there was no significant difference between groups (P>0.05); eGFR of treatment group increased compared with before treatment (P<0.05), and eGFR of treatment group was significantly higher than that of control group (P<0.05); Alb of treatment group increased compared with before treatment (P<0.05), and there was no significant difference between groups (P>0.05). See Tables 8-10.

[0143] Table 8 Comparison of Scr levels of two groups (μmol / L)

[0144]

[0145] Note: ** P<0.01; * P<0.05. Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0146] Table 9 Comparison of BUN levels of two groups (mmol / L)

[0147]

[0148] Note: ** P<0.01; * P<0.05. Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0149] Table 10 Comparison of eGFR levels of two groups (ml / min / 1.73m 2 )

[0150]

[0151] Note: ** P<0.01; * P<0.05. Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, #P<0.05.

[0152] Table 11 Comparison of Alb levels between two groups (g / L)

[0153]

[0154] Note: ** P<0.01; * P<0.05. Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0155] 3. Comparison of immune inflammatory indicators between two groups

[0156] After treatment, IL-1β, IL-6, IL-8, TNF-α, MCP-1 and endotoxin in the treatment group were decreased compared with before treatment (P<0.05), and were significantly lower than those in the control group (P<0.05); IL-10 in the treatment group was increased compared with before treatment (P<0.05), and was significantly higher than that in the control group (P<0.05). See Tables 12-18.

[0157] Table 12 Comparison of IL-1β levels between two groups (pg / ml) [M (P25-P75)]

[0158]

[0159] Note: Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0160] Table 13 Comparison of IL-6 levels between two groups (pg / ml) [M (P25-P75)]

[0161]

[0162] Note: Comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0163] Table 14 Comparison of IL-8 levels between two groups (pg / ml) [M (P25-P75)]

[0164]

[0165] Note: comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0166] Table 15 Comparison of IL-10 levels between two groups (pg / ml) [M (P25~P75)]

[0167]

[0168] Note: comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0169] Table 16 Comparison of TNF-α levels between two groups (pg / ml) [M (P25~P75)]

[0170]

[0171] Note: comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0172] Table 17 Comparison of MCP-1 levels between two groups (pg / ml) [M (P25~P75)]

[0173]

[0174] Note: comparison between groups, compared with the control group, △△ P<0.01, △ P<0.05; comparison within groups, compared with before treatment, ## P<0.01, # P<0.05.

[0175] Table 18 Comparison of endotoxin levels between two groups (pg / ml) [M (P25~P75)]

[0176]

[0177] Note: comparison between groups, compared with the control group, △△ P<0.01, △P<0.05; comparison within group, compared with before treatment, ## P<0.01, # P<0.05.

[0178] The preferred embodiments of the present application have been disclosed with the specific details of the application to provide complete understanding of the application. However, it will be apparent to those skilled in the art that various changes in form and details can be made to the application without departing from the spirit and scope of the application. Such equivalent changes in form and details are intended to be included in the scope of the application defined by the appended claims.

Claims

1. A traditional Chinese medicine composition for treating IgA nephropathy, characterized in that, The raw medicinal materials are prepared by the following weight parts: Radix Codonopsis 9-15 parts, Radix Astragali 9-15 parts, dried ginger 6-9 parts, prepared Rhizoma Pinelliae 9-15 parts, Radix Scrophulariae 6-9 parts, Rhizoma Coptidis 3-6 parts, Radix Rehmanniae 15-45 parts, Radix Pyrrosiae 9-15 parts, Radix Taxilli 9-15 parts, and honey-fried Radix Glycyrrhizae 3-9 parts.

2. The traditional Chinese medicine composition for treating IgA nephropathy according to claim 1, characterized in that, The raw medicinal materials are prepared by the following weight parts: Radix Codonopsis 12-15 parts, Radix Astragali 12-15 parts, dried ginger 8-9 parts, prepared Rhizoma Pinelliae 12-15 parts, Radix Scrophulariae 8-9 parts, Rhizoma Coptidis 5-6 parts, Radix Rehmanniae 30-45 parts, Radix Pyrrosiae 12-15 parts, Radix Taxilli 12-15 parts, and honey-fried Radix Glycyrrhizae 6-9 parts.

3. The traditional Chinese medicine composition for treating IgA nephropathy according to claim 1, characterized in that, The raw medicinal materials are prepared by the following weight parts: Radix Codonopsis 15 parts, Radix Astragali 15 parts, dried ginger 9 parts, prepared Rhizoma Pinelliae 15 parts, Radix Scrophulariae 9 parts, Rhizoma Coptidis 6 parts, Radix Rehmanniae 30 parts, Radix Pyrrosiae 15 parts, Radix Taxilli 15 parts, and honey-fried Radix Glycyrrhizae 9 parts.

4. Use of the traditional Chinese medicine composition for treating IgA nephropathy according to any one of claims 1-3 in the preparation of a medicament for treating IgA nephropathy.

5. Use according to claim 4, characterized in that, The IgA nephropathy includes chronic glomerulonephritis.

6. A pharmaceutical preparation comprising the traditional Chinese medicine composition for treating IgA nephropathy according to any one of claims 1-3 as an active ingredient. The pharmaceutical preparation is prepared into a clinically acceptable conventional pharmaceutical preparation according to a conventional preparation method of traditional Chinese medicine.

7. The pharmaceutical preparation according to claim 6, characterized in that, The pharmaceutical preparation is in the form of decoction, granules, powder, capsules, tablets or mixture.

8. The pharmaceutical preparation according to claim 6, characterized in that, The pharmaceutical preparation further comprises a pharmaceutically acceptable carrier or excipient.

9. The pharmaceutical preparation according to claim 6, characterized in that, The active ingredient of the pharmaceutical preparation is prepared by water extraction or 40-80% volume percentage concentration of ethanol extraction.

10. Use of the pharmaceutical preparation according to any one of claims 6-9 in the preparation of a medicament for treating IgA nephropathy.

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