Traditional Chinese medicine composition for treating IgA nephropathy

Abstract

The invention relates to a traditional Chinese medicine composition for treating IgA nephropathy. The traditional Chinese medicine composition is prepared from 10-30 parts by weight of astragalus membranaceus, 5-25 parts by weight of codonopsis pilosula, 5-25 parts by weight of centella asiatica, 5-25 parts by weight of poria cocos, 1-20 parts by weight of herba epimedii, 1-20 parts by weight of angelica sinensis, 1-20 parts by weight of fried curcuma zedoary and 1-20 parts by weight of radix clematidis. All the medicines in the traditional Chinese medicine composition have respective advantages, are fused in one prescription and supplement each other, have the effects of tonifying spleen and kidney, clearing heat and promoting diuresis, and dredging collaterals and eliminating diseases together, and have a remarkable curative effect on treating the IgA nephropathy.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicines, and in particular to a traditional Chinese medicine composition for treating IgA nephropathy. Background Art

[0002] Immunoglobulin Anephropathy (IgAN) is the most common primary glomerulonephritis in the world and one of the main causes of chronic kidney disease and end-stage renal failure. Up to 50% of patients may progress to end-stage renal disease within 25 years after the diagnosis of IgAN, and the average life expectancy is reduced by about 10 years. The pathological characteristics of IgAN are mainly manifested by the deposition of immunoglobulin A (IgA) complexes in the glomerular mesangial area, leading to glomerular inflammation and fibrosis, which may eventually progress to end-stage renal disease. One of the core mechanisms of IgAN pathogenesis is the decreased expression levels of core 1β1,3-galactosyltransferase (Core 1β1,3-galactosyltransferase, C1GALT1) and its chaperone protein (C1GALT1-specific chaperone 1, COSMC). C1GALT1 is responsible for regulating the O-glycosylation of IgA1 molecules, and downregulation of C1GALT1 expression leads to abnormal IgA1 glycosylation, especially the production of galactose-deficient IgA1 (Gd-IgA1), which aggravates the accumulation of IgA immune complexes in the glomeruli. COSMC acts as a specific molecular chaperone of C1GALT1, helping it to fold correctly and maintain activity. Increased levels of Gd-IgA1 in serum are associated with the severity of IgAN patients.

[0003] At present, various schools of traditional Chinese medicine have not reached a consensus on the etiology, pathogenesis and syndrome differentiation of IgAN, but they have basically reached a consensus on the pathological characteristics of "primary deficiency and superficial excess". Among them, "primary deficiency" mainly refers to spleen and kidney deficiency, and "superficial excess" mainly refers to damp heat, blood stasis and heat toxicity. The most common symptoms of IgAN are hematuria and proteinuria. The hematuria can be microscopic hematuria or visible hematuria, and those with more proteinuria can reach the level of nephrotic syndrome. The spleen is the source of qi and blood biochemical transformation. There is a saying that "internal injury to the spleen and stomach will cause all kinds of diseases". Spleen deficiency cannot transport and transform the essence of water and grain, qi and blood are lacking in source, and blood is not fixed and overflows; the kidney is responsible for sealing and storage, kidney qi deficiency, sealing and storage failure, then hematuria and proteinuria will occur. The spleen is responsible for ascending the clear, and the kidney is responsible for qi transformation. If the spleen fails to ascend the clear and the kidney qi transformation is abnormal, then qi stagnation and water retention will occur, and the three burners will be unfavorable. Over time, damp heat and dampness and blood stasis will occur, which will also accelerate the development of IgAN. This shows that spleen and kidney functions influence each other and play an important role in the development of IgAN.

[0004] In recent years, domestic and foreign scholars have proposed the quadruple-hit theory to explain the pathogenesis of IgAN, including the increase of Gd-IgA1 in the circulation, the production of autoantibodies against Gd-IgA1, the deposition of IgA1-related immune complexes in the glomeruli, and the resulting inflammatory response and renal tissue damage. C1GALT1, as a key glycosyltransferase, adds galactose to the O-glycosylation of IgA1 molecules. COSMC can stabilize and activate the function of C1GALT1 to ensure the smooth progress of the glycosylation process. In IgAN, the absence or low expression of both will lead to abnormal O-glycosylation modification of IgA1 molecules and the production of a large amount of Gd-IgA1. Gd-IgA1 immune complexes are deposited in the glomerular mesangial area to activate mesangial cells, triggering cell proliferation, extracellular matrix expansion and inflammatory response, playing a core role in the pathogenesis of IgAN. B-cell activating factor (BAFF) and rat proliferation-inducing ligand (APRIL) activate differentiated B cells and promote the production and secretion of IgA1. BAFF upregulation can inhibit C1GALT1ization and COSMC expression, which is one of the key factors in the pathogenesis of IgAN. In addition, the changes in the levels of inflammatory and anti-inflammatory factors in the renal tissue of rats with IgAN model reflect the complex immune regulation process. As an important pattern recognition receptor, rat Toll-like receptor 4 (TLR4) can recognize pathogen-associated molecular patterns and activate downstream inflammatory signaling pathways. In IgAN, upregulation of TLR4 expression may promote the production of inflammatory factors by activating the nuclear factor κB signaling pathway, thereby aggravating renal inflammatory response and damage. The expression level of TNF-α is positively correlated with the severity of IgAN, which can activate the apoptosis pathway, increase tubular cell exhaustion, and promote tubular atrophy and renal fibrosis. Studies have shown that cytokines IL-6 and IL-4 increase the degree of IgA1 galactose deficiency by regulating the expression of key glycosyltransferases. IL-10 is an important anti-inflammatory cytokine, and its expression level is related to the severity of tubulointerstitial damage. Currently, IgAN treatment strategies targeting C1GALT1 and COSMC are under continuous exploration.

[0005] There is no report on effective Chinese medicine composition for treating IgA nephropathy. Summary of the invention

[0006] To solve the above technical problems, the present invention includes the following aspects:

[0007] A first aspect of the present invention provides a traditional Chinese medicine composition for treating IgA nephropathy, wherein the traditional Chinese medicine composition is prepared from astragalus, codonopsis, centella asiatica, tuckahoe, epimedium, angelica, stir-fried curcuma and clematis.

[0008] Preferably, the traditional Chinese medicine composition is prepared from 10-30 parts by weight of Astragalus, 5-25 parts by weight of Codonopsis pilosula, 5-25 parts by weight of Centella asiatica, 5-25 parts by weight of Poria, 1-20 parts by weight of Epimedium, 1-20 parts by weight of Angelica sinensis, 1-20 parts by weight of Curcuma zedoaria and 1-20 parts by weight of Clematis chinensis.

[0009] More preferably, the traditional Chinese medicine composition is prepared from 15-25 parts by weight of Astragalus, 10-20 parts by weight of Codonopsis pilosula, 10-20 parts by weight of Centella asiatica, 10-20 parts by weight of Poria, 5-15 parts by weight of Epimedium, 5-15 parts by weight of Angelica sinensis, 5-15 parts by weight of Curcuma zedoaria and 5-15 parts by weight of Clematis chinensis.

[0010] Further preferably, the traditional Chinese medicine composition is prepared from 18-22 parts by weight of Astragalus, 12-18 parts by weight of Codonopsis pilosula, 12-18 parts by weight of Centella asiatica, 12-18 parts by weight of Poria, 8-12 parts by weight of Epimedium, 8-12 parts by weight of Angelica sinensis, 8-12 parts by weight of Curcuma zedoaria and 8-12 parts by weight of Clematis chinensis.

[0011] Most preferably, the Chinese medicine composition is prepared from 20 parts by weight of Astragalus, 15 parts by weight of Codonopsis pilosula, 15 parts by weight of Centella asiatica, 15 parts by weight of Poria cocos, 10 parts by weight of Epimedium, 10 parts by weight of Angelica sinensis, 10 parts by weight of Curcuma zedoaria and 10 parts by weight of Clematis chinensis.

[0012] Preferably, the traditional Chinese medicine composition further comprises pharmaceutically acceptable excipients.

[0013] Preferably, the Chinese medicine composition is an oral preparation.

[0014] Preferably, the Chinese medicine composition is a decoction, tablet, capsule, granule or pill. More preferably, the Chinese medicine composition is a decoction.

[0015] The second aspect of the present invention provides a method for preparing the above-mentioned Chinese medicine composition, the method comprising the following steps:

[0016] (1) Weigh the prescribed amount of Astragalus, Codonopsis, Centella, Poria, Epimedium, Angelica, Curcuma and Clematis, mix them, add 2-4 times the total weight of the medicinal materials in distilled water, and soak them thoroughly;

[0017] (2) first decocting over high heat, then decocting over low heat, filtering, adding 2-4 times the total weight of distilled water to the filtered Chinese medicinal materials, first decocting over high heat, then decocting over low heat, and filtering;

[0018] (3) Combine the two filtrates and concentrate to 3-4 times the total weight of the Chinese medicinal materials.

[0019] Preferably, in step (1), distilled water in an amount 3 times the total weight of the medicinal materials is added.

[0020] Preferably, distilled water in an amount twice the total weight of the medicinal materials is added in step (2).

[0021] Preferably, in step (3), the concentration is to be 3.8 times of the total weight of the Chinese medicinal materials.

[0022] Preferably, the soaking time in step (1) is 20-40 min. More preferably, the soaking time in step (1) is 30 min.

[0023] Preferably, in step (2), the decocting is performed over high heat for 5-15 minutes each time, and then decocted over low heat for 30-60 minutes. More preferably, the decocting is performed over high heat for 10 minutes each time, and then decocted over low heat for 40 minutes.

[0024] The third aspect of the present invention provides use of the above-mentioned Chinese medicine composition in the preparation of a medicament for treating IgA nephropathy.

[0025] The technical effects produced by the present invention are:

[0026] 1. The present invention uses IgAN model rats as the research object, screens out the best Chinese medicine composition for treating IgAN (Example 1) by optimizing the selection and ratio of Chinese medicine components, and explores the specific mechanism of action of the Chinese medicine composition of the present invention for treating IgAN, providing a new strategy for the clinical treatment of IgAN.

[0027] 2. In the Chinese medicine formula of the present invention, astragalus and codonopsis pilosula can strengthen the spleen and replenish qi, and help the acquired foundation; celery thorn can nourish the kidney and benefit the essence, and strengthen the innate foundation; angelica and curcuma can promote qi and blood circulation, and remove blood stasis and dredge the meridians; Poria, Centella asiatica, and clematis can strengthen the spleen and eliminate dampness, and clear away heat and detoxify. All the medicines show their strengths, and they are integrated into one prescription, complementing each other, and playing the effects of nourishing the spleen and kidney, clearing away heat and dampness, and dredging the meridians and eliminating symptoms. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is the IgA content of rat kidneys in each test group (400×), wherein A is the negative control group, B is the model group, C is the positive control group, D is the low-dose group of the Chinese medicine composition of the present invention, and E is the high-dose group of the Chinese medicine composition of the present invention;

[0029] Figure 2 is the serum IgA1, Gd-IgA1, BAFF and APRIL levels of rats in each test group, wherein A is a negative control group, B is a model group, C is a positive control group, D is a low-dose group of the Chinese medicine composition of the present invention, and E is a high-dose group of the Chinese medicine composition of the present invention;

[0030] Figure 3are the UTP, Cr, and BUN levels of rats in each test group, wherein A is a negative control group, B is a model group, C is a positive control group, D is a low-dose group of the Chinese medicine composition of the present invention, and E is a high-dose group of the Chinese medicine composition of the present invention;

[0031] Figure 4 The pathological morphology of the kidneys of rats in each test group (HE staining, 400×), wherein A is the negative control group, B is the model group, C is the positive control group, D is the low-dose group of the Chinese medicine composition of the present invention, and E is the high-dose group of the Chinese medicine composition of the present invention;

[0032] Figure 5 is the level of TLR4, TNF-α, IL-6, IL-4, IL-10 and INF-r in the kidney of rats in each test group, wherein A is the negative control group, B is the model group, C is the positive control group, D is the low-dose group of the Chinese medicine composition of the present invention, and E is the high-dose group of the Chinese medicine composition of the present invention;

[0033] Figure 6 is the relative mRNA expression of C1galt1 and Cosmc in the kidney of rats in each test group, wherein A is the negative control group, B is the model group, C is the positive control group, D is the low-dose group of the Chinese medicine composition of the present invention, and E is the high-dose group of the Chinese medicine composition of the present invention;

[0034] Figure 7 The expression of C1GALT1 and COSMC proteins in the kidney of rats in each test group, wherein A is the negative control group, B is the model group, C is the positive control group, D is the low-dose group of the Chinese medicine composition of the present invention, and E is the high-dose group of the Chinese medicine composition of the present invention.

[0035] Unless otherwise specified, the meanings of the statistical difference symbols * and # in the accompanying drawings of the present invention are as follows: compared with the control group, *** represents P < 0.005 for the Chinese medicine composition group of the present invention; compared with the model group, # represents P < 0.05, ## represents P < 0.01, and ### represents P < 0.005. DETAILED DESCRIPTION

[0036] Example 1

[0037] Weigh 20g of Astragalus, 15g of Codonopsis, 15g of Centella asiatica, 15g of Poria, 10g of Epimedium, 10g of Angelica, 10g of Rhizoma Curcumae and 10g of Radix Clematis, add 3 times of distilled water of the total weight of the medicinal materials after mixing, soak for 30min, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter, add 2 times of distilled water of the total weight of the medicinal materials after filtering, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter. Combine the two filtrates and concentrate them into 400mL to obtain Chinese medicine decoction 1 of the present invention.

[0038] Example 2

[0039] Weigh 10g of Astragalus, 20g of Codonopsis, 20g of Centella asiatica, 15g of Poria, 10g of Epimedium, 10g of Angelica, 10g of Rhizoma Curcumae and 10g of Radix Clematis, add 3 times of distilled water of the total weight of the medicinal materials after mixing, soak for 30min, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter, add 2 times of distilled water of the total weight of the medicinal materials after filtering, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter. Combine the two filtrates and concentrate them into 400mL to obtain Chinese medicine decoction 2 of the present invention.

[0040] Example 3

[0041] Weigh 10g of Astragalus, 15g of Codonopsis, 15g of Centella asiatica, 15g of Poria, 15g of Epimedium, 15g of Angelica, 10g of Rhizoma Curcumae and 10g of Radix Clematis, add 3 times of distilled water of the total weight of the medicinal materials after mixing, soak for 30min, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter, add 2 times of distilled water of the total weight of the medicinal materials after filtering, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter. Combine the two filtrates and concentrate them into 400mL to obtain Chinese medicine decoction 3 of the present invention.

[0042] Example 4

[0043] Weigh 10g of Astragalus, 15g of Codonopsis, 15g of Centella asiatica, 15g of Poria, 10g of Epimedium, 10g of Angelica, 15g of Rhizoma Curcumae and 15g of Radix Clematis, add 3 times of distilled water of the total weight of the medicinal materials after mixing, soak for 30min, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter, add 2 times of distilled water of the total weight of the medicinal materials after filtering, first decocted with high heat for 10min, then decocted with slow fire for 40min, filter. Combine the two filtrates and concentrate them into 400mL to obtain Chinese medicine decoction 4 of the present invention.

[0044] Experimental Example 1: Study on the therapeutic effect and mechanism of the Chinese medicine composition of the present invention on IgA nephropathy model rats

[0045] 1. Test methods

[0046] 1.1 Reagents and instruments

[0047] Benazepril hydrochloride was purchased from Beijing Solebow Technology Co., Ltd., lipopolysaccharide was purchased from Beijing Solebow Technology Co., Ltd., carbon tetrachloride was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., castor oil was purchased from Beijing Solebow Technology Co., Ltd., rat proliferation-inducing ligand (APRIL) detection kit was purchased from RayBiotech, USA, rat IgA1, Gd-lgA1, B-cell activating factor (BAFF) detection kit was purchased from Shanghai Jining Biotechnology Co., Ltd., rat Toll-like receptors 4 (TLR4), tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-4, IL-10, interferon-gamma (IFN-γ) detection kit was purchased from Wuhan Elaruite Biotechnology Co., Ltd., UTP, blood urea nitrogen ( Nitrogen, BUN) and Cr detection kits were purchased from Nanjing Jiancheng Biotechnology Co., Ltd., eosin staining solution was purchased from Beijing Solebow Technology Co., Ltd., hematoxylin staining solution was purchased from Beijing Regen Biotechnology Co., Ltd., IgA antibody was purchased from Abcam (Shanghai) Trading Co., Ltd., FITC-labeled rabbit anti-goat IgG was purchased from Beijing Solebow Technology Co., Ltd., C1GALT1 antibody was purchased from Wuhan Biooute Biotechnology Co., Ltd., COSMC antibody was purchased from Wuhan Sanying Biotechnology Co., Ltd., total RNA extraction reagent was purchased from Beijing Solebow Technology Co., Ltd., reverse transcription kit was purchased from Beijing Quanshijin Biotechnology Co., Ltd., and amplification kit was purchased from Beijing Quanshijin Biotechnology Co., Ltd.

[0048] The fluorescence microscope was purchased from Nikon Instrument (Shanghai) Co., Ltd. (model: ECLIPSE Ni), the ELISA detector was purchased from Berton Instrument Co., Ltd. (model: SYNERGYH1), the chemiluminescence analyzer was purchased from Shanghai Tianneng Technology Co., Ltd. (model: 6300), the fluorescence quantitative PCR instrument was purchased from Shanghai Hongshi Medical Technology Co., Ltd. (model: SLAN-965), and the ultra-micro spectrophotometer was purchased from Wuyi Technology Co., Ltd. (model: OneDrop).

[0049] 1.2. Animal model establishment, grouping and drug administration

[0050] Twenty-five 8-week-old SPF male SD rats were selected and divided into a negative control group (n=5) and a model group (n=20) according to the random number table method. The rats were adaptively raised in the laboratory cage for 1 week. The feeding conditions were maintained at 22±1℃ and 65-70% humidity. The experimental IgAN model of rats was established by oral immunogen + subcutaneous injection of carbon tetrachloride + intravenous injection of lipopolysaccharide. The model group was gavaged with bovine serum albumin 400 mg / kg every other day from the first day of the experiment for 6 weeks; subcutaneous injection of castor oil 0.5 mL + carbon tetrachloride 0.1 mL once a week for 8 weeks; 0.5 mg lipopolysaccharide was injected into the tail vein in the 6th and 8th weeks; the modeling time was 8 weeks. The negative control group was gavaged with distilled water instead of 0.9% saline by tail vein injection. After the model was successfully prepared, the model group rats were divided into model group (n=5), benazepril group (n=5), low-dose group of the Chinese medicine composition of the present invention (n=5), and high-dose group of the Chinese medicine composition of the present invention (n=5) according to the random number expression method. Take 800mL of the decoction prepared by the method of Example 1, concentrate to 200mL and 50mL respectively, as the dosing liquid of the low-dose group and the high-dose group (the dosing liquid is insufficient during the experiment, and it is prepared according to the above method). The rats in the low-dose group and the high-dose group of the Chinese medicine composition of the present invention were gavaged with 2mL of the above-prepared liquid. The benazepril group was given a suspension of benazepril hydrochloride and distilled water, and each rat was gavaged with 1.8mg of benazepril daily, and the negative control group and the model group were gavaged with the same amount of distilled water. Once a day, for 4 consecutive weeks.

[0051] 1.3. Test index detection

[0052] 1.3.1 Serum biochemistry and renal tissue samples

[0053] After the last administration, serum was collected from rats in each experimental group after intraperitoneal anesthesia, and serum IgA1, Gd-lgA1, BAFF, APRIL, Cr and BUN levels were detected by enzyme-linked immunosorbent assay (ELISA). The bilateral kidneys of the mice were removed, and the cortex of the left kidney was separated for fixation or frozen section; the cortex of the right kidney was separated for liquid nitrogen freezing. After renal tissue homogenization, ElLISA was used to detect the levels of TLR4, TNF-α, IL-6, IL-4, IL-10 and IFN-γ in renal tissue.

[0054] 1.3.2 Quantitative detection of 24-hour urine protein

[0055] Under a sterile environment, rat urine was collected in a metabolic cage, the supernatant was obtained by centrifugation, and the urine protein content was detected using a UTP detection kit.

[0056] 1.3.3 Renal tissue pathological observation

[0057] After dehydration, wax embedding, paraffin sectioning, and baking, the renal tissue was dewaxed and hydrated after being soaked in xylene and gradient alcohol, and the sections were routinely stained with hematoxylin-eosin (HE), and the pathological changes of renal tissue were observed under a microscope. After sectioning, dewaxing and hydration, PBS washing, and BSA blocking, the renal tissue was incubated with IgA antibody as the primary antibody, and FITC-labeled rabbit anti-goat IgG was added for secondary antibody incubation, PBS washing, anti-fluorescence quenching and sealing, and the deposition site and intensity of IgA in the renal tissue of each group of rats were observed under a fluorescence microscope, and the semi-quantitative standard of the immunofluorescence intensity 5-level method commonly used at home and abroad was used.

[0058] 1.3.4. Renal tissue protein extraction and Western Blot detection

[0059] The kidney tissue was homogenized according to the conventional method; the protein concentration of the sample was determined using a BCA protein quantification kit, followed by electrophoresis, transfer to the membrane, blocking with 5% skim milk powder for 1 hour, adding the primary antibody (1:1000) and incubating overnight at 4°C, adding the HRP-labeled secondary antibody (1:5000) and incubating at room temperature for 2 hours, washing the membrane and adding the ECL chemiluminescent reagent for color development, using the ChemiDocMP imaging system to acquire images, and using ImageJ image analysis software for semi-quantitative analysis.

[0060] 1.3.5 Real-time quantitative polymerase chain reaction (RT-qPCR) detection

[0061] Total RNA from renal tissue was extracted according to the instructions of RNA extraction kit. Total RNA was reverse transcribed using the full-gold reverse transcription kit, and the PCR reaction system was configured using the full-gold amplification kit for quantitative PCR detection. The primer sequences are shown in Table 1. The qPCR reaction program was 94°C, 30s; 94°C, 5s; 60°C, 30s; 40 cycles, with β-actin as the internal reference, and the 2–ΔΔCT method was used to analyze the expression of each gene.

[0062] Table 1 Primer sequences

[0063]

[0064] 1.4 Statistical analysis

[0065] GraphPad Prism 10.0 software was used for data analysis and plotting. The independent sample t test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. P < 0.05 indicated that the difference was statistically significant.

[0066] 2. Test results

[0067] 2.1 Comparison of changes in renal IgA content in rats of each group

[0068] The results of immunofluorescence staining showed that compared with the negative control group, branched and granular IgA deposits were observed in the glomerular mesangial area of ​​the model group, indicating that the experimental IgAN model of rats was successfully established. Compared with the model group, the IgA content in the renal tissue of rats in the low-dose and high-dose administration groups was significantly reduced, indicating that the Chinese medicine composition of the present invention can inhibit the deposition of IgA in the kidney. The inhibitory effect of the high-dose group on the IgA deposition kidney was consistent with that of the benazepril positive drug group (see Figure 1 ).

[0069] 2.2 Comparison of serum IgA1-related immune factors in rats of each group

[0070] The levels of IgA1, Gd-IgA1, BAFF and APRIL in rat serum were detected by ELISA. It was found that compared with the negative control group, the levels of IgA1, Gd-IgA1, BAFF and APRIL in the serum of the rats in the model group were significantly increased (P < 0.005). The Chinese medicine composition of the present invention can significantly reduce the levels of IgA1, Gd-IgA1, BAFF and APRIL. The inhibition rates of IgA1, Gd-IgA1, BAFF and APRIL in the high-dose administration group were 54.0% (P < 0.005), 48.1% (P < 0.005), 45.9% (P < 0.005) and 38.5% (P < 0.005), respectively, which were equivalent to the intervention effect of the benazepril positive drug intervention group (see Figure 2 ). The above test results show that the Chinese medicine composition of the present invention can significantly reduce the level of IgA1 related immune factors in serum.

[0071] 2.3 Comparison of renal function among rats in each group

[0072] UTP, Cr and BUN are used as indicators to evaluate renal damage in IgAN. The UTP, Cr and BUN of the model group rats were significantly higher than those of the negative control group (P < 0.005). Compared with the model group, the UTP, Cr and BUN of the rats intervened with the Buxu Qushi Xiaozheng recipe decreased, which was statistically significant. The inhibition rate of the high-dose Buxu Qushi Xiaozheng recipe intervention group was equivalent to that of the benazepril positive drug intervention group, and the inhibition rates of UTP, Cr and BUN reached 76.0% (P < 0.005), 51.1% (P < 0.01) and 44.0% (P < 0.005), respectively. The above shows that the Buxu Qushi Xiaozheng recipe can significantly improve the renal function of IgAN rats. (See Figure 3 ). The above test results show that the Chinese medicine composition of the present invention can significantly improve the renal function of IgAN rats.

[0073] 2.4 Comparison of renal pathological changes in rats in each group

[0074] HE staining showed that compared with the negative control group, the model group showed pathological changes of mesangial cell proliferation and increased mesangial matrix to varying degrees, and some capillary loops were compressed, lumen stenosis and balloon adhesion were observed. Compared with the model group, the glomerular and renal interstitial lesions of rats treated with the Chinese medicine composition of the present invention and the positive drug were alleviated to varying degrees, indicating that the Chinese medicine composition of the present invention can improve the renal tissue lesions of IgAN rats (see Figure 4 ).

[0075] 2.5 Comparison of renal inflammatory factors in rats of each group

[0076] The levels of TLR4, TNF-α, IL-6, IL-4, IL-10, and IFN-γ in the kidneys of rats were detected by ELISA. It was found that compared with the negative control group, the levels of inflammatory factors TLR4, TNF-α, IL-6, and IL-4 in the model group were significantly increased (P < 0.005), while the levels of anti-inflammatory factors IL-10 and IFN-γ were significantly decreased (P < 0.005). After treatment with the Chinese medicine composition of the present invention, the levels of TLR4, TNF-α, IL-6, and IL-4 decreased significantly, and the levels of IL-10 and INF-r increased significantly (see Figure 5 ). The above test results show that the Chinese medicine composition of the present invention can significantly inhibit the inflammatory response of the kidney of IgAN rats.

[0077] 2.6 Comparison of mRNA expression of C1GALT1 and COSMC in rat kidneys among the groups

[0078] The decreased activity of C1GALT1 and its partner protein COSMC is one of the pathogenesis of IgAN. RT-qPCR detection showed that the gene expression levels of C1GALT1 and COSMC in the kidneys of rats in the model group were reduced by 95.5% (P < 0.005) and 92.0% (P < 0.005) respectively compared with those in the negative control group. Compared with the model group, the gene expression levels of C1GALT1 and COSMC in rats given the Chinese medicine composition of the present invention were significantly upregulated, among which the gene expression levels of C1GALT1 and COSMC in the high-dose group of the Chinese medicine composition of the present invention were increased by 7.2 times and 6.5 times respectively (see Figure 6 ).

[0079] 2.7 Comparison of protein expression of C1GALT1 and COSMC in kidney of rats in each group

[0080] Western Blot and semi-quantitative analysis results showed that compared with the negative control group, the expression levels of C1GALT1 and COSMC proteins in the model group decreased by 61.9% (P < 0.005) and 63.3% (P < 0.005), respectively. After treatment with the Chinese medicine composition of the present invention, the expression levels of C1GALT1 and COSMC proteins were significantly upregulated. The expression levels of C1GALT1 and COSMC proteins in the high-dose group of the Chinese medicine composition of the present invention were upregulated by 2.5 times and 2.2 times, respectively (see Figure 7 The above test results show that the Chinese medicine composition of the present invention may inhibit the production and deposition of IgA1-related immune complexes and alleviate renal tissue damage mediated by immune complexes by restoring the expression of C1GALT1 and its chaperone protein COSMC in the kidneys of IgAN rats.

[0081] In summary, the Chinese medicine composition of the present invention has significant therapeutic effects in reducing renal IgA deposition in IgAN model rats, reducing serum IgA1-related immune factor levels, inhibiting renal inflammatory response, improving renal function and alleviating renal lesions. It is confirmed that the Chinese medicine composition of the present invention may reduce the immune complex-mediated renal tissue injury mechanism by regulating the expression of C1GALT1 and its partner protein COSMC. It is expected that the Chinese medicine composition of the present invention can provide an effective clinical treatment plan for IgAN patients.

[0082] Although specific embodiments of the present invention have been described, it will be appreciated by those skilled in the art that various changes and modifications may be made to the present invention without departing from the scope or spirit of the present invention. Therefore, the present invention is intended to cover all such changes and modifications that fall within the scope of the appended claims and their equivalents.

Claims

1. A Chinese medicine composition for treating IgA nephropathy, characterized in that: The traditional Chinese medicine composition is prepared from astragalus, codonopsis, centella asiatica, tuckahoe, epimedium, angelica, stir-fried curcuma and clematis.

2. The Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared from 10-30 parts by weight of astragalus, 5-25 parts by weight of codonopsis pilosula, 5-25 parts by weight of centella asiatica, 5-25 parts by weight of tuckahoe, 1-20 parts by weight of epimedium, 1-20 parts by weight of angelica sinensis, 1-20 parts by weight of stir-fried curcuma and 1-20 parts by weight of clematis.

3. The Chinese medicine composition according to claim 2, characterized in that: The traditional Chinese medicine composition is prepared from 15-25 parts by weight of astragalus, 10-20 parts by weight of codonopsis pilosula, 10-20 parts by weight of centella asiatica, 10-20 parts by weight of tuckahoe, 5-15 parts by weight of epimedium, 5-15 parts by weight of angelica sinensis, 5-15 parts by weight of stir-fried curcuma and 5-15 parts by weight of clematis.

4. The Chinese medicine composition according to claim 3, characterized in that: The traditional Chinese medicine composition is prepared from 18-22 parts by weight of astragalus, 12-18 parts by weight of codonopsis pilosula, 12-18 parts by weight of centella asiatica, 12-18 parts by weight of tuckahoe, 8-12 parts by weight of epimedium, 8-12 parts by weight of angelica sinensis, 8-12 parts by weight of stir-fried curcuma and 8-12 parts by weight of clematis.

5. The Chinese medicine composition according to claim 4, characterized in that: The traditional Chinese medicine composition is prepared from 20 parts by weight of astragalus, 15 parts by weight of codonopsis, 15 parts by weight of centella asiatica, 15 parts by weight of tuckahoe, 10 parts by weight of epimedium, 10 parts by weight of angelica, 10 parts by weight of stir-fried curcuma and 10 parts by weight of clematis.

6. The Chinese medicine composition according to any one of claims 1 to 5, characterized in that: The traditional Chinese medicine composition further includes pharmaceutically acceptable excipients.

7. The Chinese medicine composition according to claim 6, characterized in that: The Chinese medicine composition is an oral preparation.

8. The method for preparing the Chinese medicine composition according to any one of claims 1 to 7, characterized in that: The method comprises the following steps: (1) Weigh the prescribed amount of Astragalus, Codonopsis, Centella, Poria, Epimedium, Angelica, Curcuma and Clematis, mix them, add 2-4 times the total weight of the medicinal materials in distilled water, and soak them thoroughly; (2) first decocting over high heat, then decocting over low heat, filtering, adding 2-4 times the total weight of distilled water to the filtered Chinese medicinal materials, first decocting over high heat, then decocting over low heat, and filtering; (3) Combine the two filtrates and concentrate to 3-4 times the total weight of the Chinese medicinal materials.

9. The preparation method according to claim 8, characterized in that: In the step (2), the mixture is boiled over high heat for 5-15 minutes each time, and then boiled over low heat for 30-60 minutes.

10. Use of the Chinese medicine composition according to any one of claims 1 to 7 in the preparation of a medicament for treating IgA nephropathy.

Images