Traditional Chinese medicine composition for treating renal proteinuria as well as preparation method and application thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating renal proteinuria as well as a preparation method and application thereof, and particularly belongs to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 30-60 parts of radix astragali, 10-20 parts of roasted rhizoma atractylodis macrocephalae, 8-16 parts of rhizoma polygoni cuspidati, 8-16 parts of rhizoma alismatis, 10-20 parts of rhizoma smilacis glabrae, 8-16 parts of rhizoma curcumae longae, 8-16 parts of lumbricus, 8-16 parts of silkworm larva, 10-20 parts of herba lysimachiae, 10-20 parts of herba centellae, 10-20 parts of sticktight and 10-20 parts of limonium bicolor. The traditional Chinese medicine composition can effectively relieve proteinuria, protect the renal function and regulate the expression of kidney inflammation macrophages, and particularly has the effects of promoting the expression of anti-inflammatory macrophages, reducing the inflammatory factor level of kidney tissue and inhibiting an inflammation signal channel. The traditional Chinese medicine composition provides a safe and effective medicine for treating proteinuria for patients.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for treating renal proteinuria, a preparation method thereof and an application thereof. Background Art

[0002] Proteinuria is an important clinical manifestation of chronic kidney disease and a marker of kidney damage. Proteinuria is associated with not only early kidney diseases but also benign diseases. Microalbuminuria is the earliest indicator of kidney complications in diabetes, obesity and metabolic syndrome. The appearance of proteinuria indicates an increased risk of secondary kidney damage such as hypertension and cardiovascular diseases. Therefore, it is an important basis for measuring the severity of the disease, clarifying the treatment plan and judging the prognosis clinically. Proteinuria is one of the important indicators for diagnosing kidney diseases and can also be an independent factor in the development of kidney diseases. Therefore, it is very meaningful to delay or prevent the occurrence of proteinuria.

[0003] Clinically, for both primary and secondary kidney diseases, many exact pathogenic mechanisms are still unclear, and there is a lack of effective therapeutic drugs in clinical practice. For example, in IgA nephropathy, studies have shown that some IgAN patients will eventually develop renal failure after long-term renal insufficiency; although there are many types of drugs for treating membranous nephropathy, there are many adverse reactions and complications, and the long-term effect is not ideal; for diabetic nephropathy patients entering the stage of massive proteinuria, due to the influence of blood sugar, the application of hormones and immunosuppressants is restricted, and there is no effective treatment method. Therefore, as an efficacy evaluation index, clinical studies have proved that controlling the level of proteinuria can effectively delay the progression of kidney diseases. Therefore, it is of great significance to find new and safe prevention strategies to control proteinuria and delay the progression of chronic kidney diseases. Summary of the Invention

[0004] In view of the current lack of effective drugs for treating proteinuria clinically, the present invention provides a traditional Chinese medicine composition for treating renal proteinuria, a preparation method thereof and an application thereof. The traditional Chinese medicine composition can treat renal proteinuria and can achieve the treatment of chronic kidney disease.

[0005] The present invention provides a traditional Chinese medicine composition, comprising the following raw materials for preparation in parts by weight: 30 - 60 parts of Astragalus membranaceus, 10 - 20 parts of stir-fried Atractylodes macrocephala, 8 - 16 parts of Polygonum cuspidatum, 8 - 16 parts of Alisma orientale, 10 - 20 parts of Smilax glabra, 8 - 16 parts of Curcuma longa, 8 - 16 parts of Pheretima aspergillum, 8 - 16 parts of Bombyx batryticatus, 10 - 20 parts of Tripterygium regelii, 10 - 20 parts of Centella asiatica, 10 - 20 parts of Bidens pilosa and 10 - 20 parts of Limonium bicolor.

[0006] Preferably, the raw materials for preparation include the following parts by weight: 30 parts of Astragalus membranaceus, 15 parts of stir-fried Atractylodes macrocephala, 12 parts of Polygonum cuspidatum, 12 parts of Alisma orientale, 15 parts of Smilax glabra, 12 parts of Curcuma longa, 12 parts of Pheretima aspergillum, 12 parts of Bombyx batryticatus, 15 parts of Tripterygium regelii, 15 parts of Centella asiatica, 15 parts of Bidens pilosa and 15 parts of Limonium bicolor.

[0007] The present invention also provides a preparation method of the traditional Chinese medicine composition described in the above technical solution, including the following steps:

[0008] Mix the raw materials with water, soak, decoct, and concentrate to obtain the traditional Chinese medicine composition.

[0009] Preferably, the soaking time is 1.5 - 2.5 h; the number of decocting times is 2 - 3 times; after concentration, the crude drug concentration is 1 - 2.5 g / mL.

[0010] The present invention also provides a traditional Chinese medicine preparation made from the traditional Chinese medicine composition described in the above technical solution.

[0011] Preferably, the dosage form of the traditional Chinese medicine preparation includes capsules, granules, medicated ointments or cut crude drugs.

[0012] The present invention also provides the application of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition prepared by the preparation method described in the above technical solution or the traditional Chinese medicine preparation described in the above technical solution in the preparation of drugs for treating chronic kidney disease.

[0013] Preferably, the chronic kidney disease includes any one or more of IgA nephropathy, membranous nephropathy, nephrotic syndrome, diabetic nephropathy and chronic nephritis.

[0014] The present invention also provides the application of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition prepared by the preparation method described in the above technical solution or the traditional Chinese medicine preparation described in the above technical solution in the preparation of drugs for treating renal proteinuria.

[0015] The present invention also provides the application of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition prepared by the preparation method described in the above technical solution or the traditional Chinese medicine preparation described in the above technical solution in the preparation of drugs having any one or more of the following effects:

[0016] ①Improving renal function;

[0017] ②Reducing the level of renal tissue inflammatory factors;

[0018] ③Promoting the transformation of renal macrophages into an anti-inflammatory phenotype;

[0019] ④Inhibiting the gene and protein expression of related molecules in the inflammatory signaling pathway TRAF6 / NF-KBp65.

[0020] The present invention provides a traditional Chinese medicine composition. The raw materials are derived from common Chinese herbal medicines, with rich sources and low prices. Based on the etiology and pathogenesis of traditional Chinese medicine and syndrome differentiation and treatment, following the principle of "invigorating the spleen and promoting diuresis, expelling wind and removing dampness", it achieves the effect of treating proteinuria. The traditional Chinese medicine composition of the present invention can effectively improve clinical symptoms such as edema and proteinuria in patients clinically. Further experimental studies confirm that the traditional Chinese medicine composition of the present invention can effectively reduce proteinuria in mice and protect renal function; its mechanism includes regulating the expression of inflammatory macrophages in the kidney, promoting the expression of anti-inflammatory macrophages, reducing the level of inflammatory factors in renal tissue, and inhibiting the role of inflammatory signaling pathways. The traditional Chinese medicine composition of the present invention provides a safe and effective drug for treating proteinuria for patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0022] Figure 1 It is a graph showing the liver function test results of Jian XX provided by the present invention;

[0023] Figure 2 It is a graph showing the 24-hour urinary protein quantification results of Jian XX provided by the present invention;

[0024] Figure 3 It is a graph showing the 24-hour urinary protein quantification test results of Wang XX provided by the present invention;

[0025] Figure 4 It is a graph showing the 24-hour urinary protein quantification test results of Yang XX provided by the present invention;

[0026] Figure 5 It is a graph showing the 24-hour urinary protein quantification test results of Wang XX provided by the present invention;

[0027] Figure 6 It is a graph showing the 24-hour urinary protein quantification test results of Li XX provided by the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0028] The present invention provides a traditional Chinese medicine composition, which includes the following raw materials for preparation in parts by weight: 30 - 60 parts of Astragalus membranaceus, 10 - 20 parts of stir-fried Atractylodes macrocephala, 8 - 16 parts of Polygonum cuspidatum, 8 - 16 parts of Alisma orientale, 10 - 20 parts of Smilax glabra, 8 - 16 parts of Curcuma longa, 8 - 16 parts of Pheretima aspergillum, 8 - 16 parts of Bombyx batryticatus, 10 - 20 parts of Tripterygium regelii, 10 - 20 parts of Centella asiatica, 10 - 20 parts of Bidens pilosa, and 10 - 20 parts of Limonium bicolor.

[0029] The present invention does not have special limitations on the sources of various traditional Chinese medicine raw materials, and conventional commercially available products well-known to those skilled in the art can be used.

[0030] The traditional Chinese medicine composition of the present invention includes 30 to 60 parts of Astragalus membranaceus. In specific embodiments, it includes 30 parts, 40 parts, 50 parts or 60 parts of Astragalus membranaceus. In the traditional Chinese medicine composition of the present invention, the function of Astragalus membranaceus is to increase plasma albumin, reduce urinary protein, inhibit the production of lipid peroxides, counteract oxygen free radical damage, promote water and sodium excretion, improve the hypercoagulable state of blood, regulate the body's immunity and reduce renal tissue damage, and protect renal function.

[0031] The traditional Chinese medicine composition of the present invention includes 10 to 20 parts of stir-fried Atractylodes macrocephala. In specific embodiments, it includes 10 parts, 12 parts, 15 parts, 18 parts or 20 parts of stir-fried Atractylodes macrocephala. In the traditional Chinese medicine composition of the present invention, the function of stir-fried Atractylodes macrocephala is to invigorate the spleen and replenish qi, regulate immunity, transform and transport dampness, induce diuresis and reduce swelling, and have anti-inflammatory and antibacterial effects.

[0032] The traditional Chinese medicine composition of the present invention includes 8 to 16 parts of Polygonum cuspidatum. In specific embodiments, it includes 25 parts, 27 parts, 30 parts or 35 parts of Polygonum cuspidatum. In the traditional Chinese medicine composition of the present invention, Polygonum cuspidatum has the effects of clearing heat and detoxifying, promoting diuresis and reducing jaundice, dispersing stasis and relieving pain, can improve glomerular filtration function, reduce kidney inflammation, lower serum creatinine, and reduce proteinuria.

[0033] The traditional Chinese medicine composition of the present invention includes 8 to 16 parts of Alisma orientale. In specific embodiments, it includes 25 parts, 27 parts, 30 parts or 35 parts of Alisma orientale. In the traditional Chinese medicine composition of the present invention, the function of Alisma orientale is to clear heat and promote diuresis, relieve stranguria and reduce swelling, reduce lipid, have anti-inflammatory and antioxidant effects, and induce diuresis.

[0034] The traditional Chinese medicine composition of the present invention includes 10 to 20 parts of Smilax glabra. In specific embodiments, it includes 10 parts, 12 parts, 15 parts, 18 parts or 20 parts of Smilax glabra. In the traditional Chinese medicine composition of the present invention, the function of Smilax glabra is to clear heat and detoxify, strengthen the spleen and eliminate dampness, and have anti-inflammatory and antibacterial effects.

[0035] The traditional Chinese medicine composition of the present invention includes 8 to 16 parts of Curcuma longa. In specific embodiments, it includes 8 parts, 10 parts, 12 parts or 16 parts of Curcuma longa. In the traditional Chinese medicine composition of the present invention, the function of Curcuma longa is to have anti-inflammatory and antioxidant effects, resist ischemia, resist tissue damage, dispel wind and remove dampness, and promote blood circulation and remove stasis.

[0036] The traditional Chinese medicine composition of the present invention includes 8 to 16 parts of Pheretima aspergillum. In specific embodiments, it includes 8 parts, 10 parts, 12 parts or 16 parts of Pheretima aspergillum. In the traditional Chinese medicine composition of the present invention, the function of Pheretima aspergillum is to have anti-inflammatory and antioxidant effects, regulate immunity, resist thrombosis, reduce blood pressure and lipid, and improve microcirculation.

[0037] The traditional Chinese medicine composition described in the present invention comprises 8 to 16 parts of Bombyx Batryticatus. In specific embodiments, it comprises 8 parts, 10 parts, 12 parts or 16 parts of Bombyx Batryticatus. In the traditional Chinese medicine composition described in the present invention, the functions of Bombyx Batryticatus are anti-inflammatory and immunomodulatory, anticoagulant, antithrombotic and antioxidant.

[0038] The traditional Chinese medicine composition described in the present invention comprises 10 to 20 parts of Schisandra Sphenanthera Rehd. et Wils. In specific embodiments, it comprises 10 parts, 12 parts, 15 parts, 18 parts or 20 parts of Schisandra Sphenanthera Rehd. et Wils. In the traditional Chinese medicine composition described in the present invention, the functions of Schisandra Sphenanthera Rehd. et Wils. are dispelling wind and removing dampness, dredging the channels and collaterals, anti-inflammatory and analgesic, immunomodulatory and antioxidant.

[0039] The traditional Chinese medicine composition described in the present invention comprises 10 to 20 parts of Centella Asiatica (L.) Urb. In specific embodiments, it comprises 10 parts, 12 parts, 15 parts, 18 parts or 20 parts of Centella Asiatica (L.) Urb. In the traditional Chinese medicine composition described in the present invention, the functions of Centella Asiatica (L.) Urb. are anti-inflammatory, antioxidant, anti-fibrotic, antibacterial, diuretic and reducing proteinuria.

[0040] The traditional Chinese medicine composition described in the present invention comprises 10 to 20 parts of Bidens Pilosa L. In specific embodiments, it comprises 10 parts, 12 parts, 15 parts, 18 parts or 20 parts of Bidens Pilosa L. In the traditional Chinese medicine composition described in the present invention, the functions of Bidens Pilosa L. are anti-inflammatory, analgesic, antibacterial, clearing heat and detoxifying, activating blood circulation to dissipate stasis and immunomodulatory.

[0041] The traditional Chinese medicine composition described in the present invention comprises 10 to 20 parts of Limonium Bicolor (Bunge) Kuntze. In specific embodiments, it comprises 10 parts, 12 parts, 15 parts, 18 parts or 20 parts of Limonium Bicolor (Bunge) Kuntze. In the traditional Chinese medicine composition described in the present invention, the functions of Limonium Bicolor (Bunge) Kuntze are clearing heat and detoxifying, diuretic and detumescence, enhancing immunity, antioxidant, improving circulation, dispelling wind and removing dampness.

[0042] The traditional Chinese medicine composition described in the present invention takes Astragalus membranaceus (Fisch.) Bunge and stir-fried Atractylodes macrocephala Koidz. as the monarch drugs, which replenish qi and consolidate the exterior, strengthen the spleen and dry dampness. The compatibility of stir-fried Atractylodes macrocephala Koidz. and Astragalus membranaceus (Fisch.) Bunge can enhance the diuretic and detumescence effect of Astragalus membranaceus (Fisch.) Bunge and strengthen the spleen to treat the root cause. Polygonum cuspidatum Sieb. et Zucc., Smilax glabra Roxb., Alisma orientale (Sam.) Juz. and Curcuma longa L. are used together as the minister drugs, playing the roles of clearing heat and promoting diuresis, promoting blood circulation and removing stasis, inducing diuresis for treating stranguria and eliminating edema, targeting the marked syndrome of dampness-heat. Pheretima aspergillum (E. Perrier), Bombyx Batryticatus and Schisandra Sphenanthera Rehd. et Wils. have the effects of expelling wind, activating blood circulation and dredging collaterals, diuretic and detumescence, assisting the minister drugs to play the roles of clearing heat and promoting diuresis, expelling wind, activating blood circulation and dredging collaterals, and relieving the marked syndromes of wind-dampness and blood stasis in the kidney. Centella Asiatica (L.) Urb., Bidens Pilosa L. and Limonium Bicolor (Bunge) Kuntze are used as the assistant drugs, with diuresis and nourishing blood used together, assisting all the drugs to play the roles of clearing heat and promoting diuresis, activating blood circulation and expelling wind, and diuretic and detumescence. The whole prescription treats both the symptoms and the root cause, and together achieves the effects of reducing nephrotic edema and reducing urinary protein.

[0043] The present invention also provides a preparation method of the traditional Chinese medicine composition described in the above technical solution, comprising the following steps:

[0044] Mix each raw material with water, soak, decoct and concentrate to obtain the traditional Chinese medicine composition.

[0045] In a specific embodiment, the mass ratio of the raw materials to water for mixing is 1:(6 - 10), specifically 1:6, 1:8 or 1:10. In a specific embodiment, the soaking time is 1.5 - 2.5 h, specifically 1.5 h, 2 h or 2.5 h; the number of decocting times is 2 - 3 times; after concentration, the crude drug concentration is 1 - 2.5 g / mL, specifically 1.13 g / mL, 1.5 g / mL, 1.8 g / mL, 2.0 g / mL or 2.5 g / mL. In a specific embodiment, after the traditional Chinese medicine composition is prepared, the traditional Chinese medicine composition is stored at 4°C.

[0046] The present invention also provides a traditional Chinese medicine preparation made from the traditional Chinese medicine composition described in the above technical solution. In a specific embodiment, the dosage form of the traditional Chinese medicine preparation includes capsules, granules, ointments or cut crude drugs.

[0047] The present invention also provides the use of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition prepared by the preparation method described in the above technical solution or the traditional Chinese medicine preparation described in the above technical solution in the preparation of a drug for treating chronic kidney disease. In a specific embodiment, the chronic kidney disease includes any one or more of IgA nephropathy, membranous nephropathy, nephrotic syndrome, diabetic nephropathy and chronic nephritis.

[0048] The present invention also provides the use of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition prepared by the preparation method described in the above technical solution or the traditional Chinese medicine preparation described in the above technical solution in the preparation of a drug for treating renal proteinuria. The traditional Chinese medicine composition of the present invention can reduce the urine protein content.

[0049] The present invention also provides the use of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition prepared by the preparation method described in the above technical solution or the traditional Chinese medicine preparation described in the above technical solution in the preparation of a drug having any one or more of the functions of ① - ④:

[0050] ① Improving renal function; ② Reducing the level of renal tissue inflammatory factors; ③ Promoting the transformation of renal macrophages into an anti-inflammatory phenotype; ④ Inhibiting the gene and protein expression of molecules related to the inflammatory signaling pathway TRAF6 / NF-κBp65. In a specific embodiment, the renal tissue inflammatory factors include MCP-1, TNF-α, IL-1β and IL-6. In a specific embodiment, the genes of molecules related to the inflammatory signaling pathway TRAF6 / NF-κBp65 include TRAF6 and NF-κBp65; the proteins of molecules related to the inflammatory signaling pathway TRAF6 / NF-κBp65 include TRAF6, p-NF-κB and NF-κBp65.

[0051] To further illustrate the present invention, the following describes in detail a traditional Chinese medicine composition for treating renal proteinuria provided by the present invention, its preparation method and application in combination with embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0052] Example 1

[0053] Making an IgA nephropathy model: Eighteen SD rats were randomly divided into a control group, a model group, and a traditional Chinese medicine composition group. Bovine serum albumin (BSA) was prepared into a 100 g / L concentration with distilled water and gavaged at 400 mg / kg (4 mL / kg) every other day for 8 consecutive weeks. Castor oil (0.3 mL) and carbon tetrachloride (CCl4, 0.1 mL) were subcutaneously injected once a week for 9 consecutive weeks; lipopolysaccharide (LPS) was prepared into a 0.25 g / L concentration with 0.9% sodium chloride solution and injected intravenously at the tail at 0.25 mg / kg at the 6th and 8th weeks. The control group was given an equal amount (4 mL / kg) of distilled water, gavaged every other day for 8 weeks; 0.4 mL of sodium chloride solution was subcutaneously injected once a week for 9 weeks, and 0.25 mg / kg of sodium chloride solution was injected intravenously at the tail at the 6th week.

[0054] Weigh the following traditional Chinese medicine decoction pieces: 30 g of Astragalus membranaceus, 15 g of stir-fried Atractylodes macrocephala, 12 g of Polygonum cuspidatum, 12 g of Alisma orientale, 15 g of Smilax glabra, 12 g of Curcuma longa, 12 g of Pheretima aspergillum, 12 g of Bombyx batryticatus, 15 g of Tripterygium regelii, 15 g of Centella asiatica, 15 g of Bidens pilosa, and 15 g of Limonium bicolor.

[0055] Administration method: The quality control of the traditional Chinese medicine decoction pieces was carried out according to the standards of the Chinese Pharmacopoeia (2015 Edition). According to the following decocting method: The traditional Chinese medicine decoction pieces were soaked in advance for 2 h, then mixed with 10 times the mass of water for decocting. The number of decocting times was 2 times. The decoction liquids were combined and concentrated into a 100 ml solution, that is, a traditional Chinese medicine stock solution with a crude drug concentration of 1.8 g / mL. After cooling, it was filled into a sterilized glass medicine bottle and stored in a 4°C refrigerator for later use. The administration dose was referred to the "Pharmacological Experiment Methodology" and converted by the direct conversion method of the equivalent dose of 200 g rats and humans (60 kg). The dosage for one rat was 3.78 g / d.

[0056] The rats were gavaged with the corresponding dose of the traditional Chinese medicine composition of the present invention for 8 consecutive weeks. Samples were taken 9 weeks after the intervention ended. Relevant indicators were detected, and the results were as follows:

[0057] 1. Detection of renal function and 24-hour urinary protein quantification

[0058] The results of renal function tests in IgA nephropathy rats intervened by the traditional Chinese medicine composition group showed that: compared with the control group, the levels of blood urea nitrogen (BUN) and creatinine (Cr) in the model group rats were significantly increased, with significant differences (P<0.0001); compared with the model group, the BUN and Cr in the traditional Chinese medicine composition group were significantly decreased, with significant differences (P<0.0001); see Table 1.

[0059] Table 1 BUN (mmol / L), Cr (umol / L) in IgA nephropathy rats Results

[0060] Grouping N BUN Cr Control group 6 3.86±0.59 57.71±10.44 Model group 6 13.40±2.48**** 171.70.±10.71**** Traditional Chinese medicine composition group 6 6.24±1.20#### 85.04±6.67**####

[0061] Compared with the control group, ****p<0.0001, **P<0.01. Compared with the model group, #p<0.0001.

[0062] The results of 24-hour urinary protein quantification showed that: compared with the control group, the 24-hour urinary protein levels in the model group and the traditional Chinese medicine composition group were significantly increased, with significant differences (P<0.0001). Compared with the model group, the 24-hour urinary protein quantification in the traditional Chinese medicine composition group was significantly decreased, with significant differences (P<0.01); see Table 2.

[0063] Table 2 24-hour urinary protein quantification (mg / L) in IgA nephropathy rats Results

[0064] Grouping N 24hU-TP Control group 6 4.14±0.55 Model group 6 26.72±2.67**** Traditional Chinese medicine composition group 6 21.78±2.14****##

[0065] Note: Compared with the control group, ****p<0.0001. Compared with the model group, ##p<0.01.

[0066] 2. Detection of intestinal mucosal sIgA content by ELISA

[0067] Secretory sIgA produced by intestinal mucosa is the most important functional executor for mucosa to participate in humoral immunity. The detection results of intestinal mucosal sIgA showed that:

[0068] Detection of intestinal mucosal sIgA in IgA nephropathy rats: compared with the control group, the sIgA level in the model group was significantly increased, with significant differences (P<0.001). The sIgA level in the traditional Chinese medicine composition group increased slightly, with no significant differences. Compared with the model group, the sIgA content in the traditional Chinese medicine composition group was significantly decreased, with significant differences (P<0.01); see Table 3.

[0069] Table 3 Intestinal sIgA content (ng / mL) in IgA nephropathy rats Results

[0070] Grouping N sIgA Control group 3 2.76±0.58 Model group 3 9.71±1.69** Traditional Chinese medicine composition group 3 5.15±1.14##

[0071] Note: Compared with the control group, **P < 0.01; compared with the model group, ##P < 0.01.

[0072] 3. Detection of the levels of inflammatory factors MCP-I, TNF-α, IL-1β, and IL-6 in the renal tissue by ELISA

[0073] The ELISA detection results of the renal tissue of IgA nephropathy rats showed that: compared with the control group, the contents of MCP-1, TNF-α, IL-1β, and IL-6 in the model group were all significantly increased, with significant differences (P < 0.0001, P < 0.001, P < 0.01). Although there was an increase in the traditional Chinese medicine composition group, there was no statistical significance; compared with the model group, the contents of MCP-1, TNF-α, IL-1β, and IL-6 in the traditional Chinese medicine composition group were all significantly decreased, with significant differences (p < 0.0001, P < 0.001, P < 0.05, P < 0.01); see Table 4.

[0074] Table 4 Contents of MCP-I, TNF-α, IL-1β, and IL-6 in the renal tissue of IgA nephropathy rats

[0075]

[0076] Note: Compared with the control group, ****p < 0.0001, ***P < 0.001, **P < 0.01; compared with the IgA group, #p < 0.0001; P < 0.001, ##P < 0.01.

[0077] 4. Detection of the mRNA of macrophage markers CD68, iNOS, CD163, and Arg-1 in the kidney by q-RTPCR

[0078] Detection of the expression of M1 macrophage markers CD68 and iNOS and M2 macrophage markers CD163 and Arg-1 mRNA in the renal tissue of IgA nephropathy rats. The research results showed that:

[0079] Compared with the control group, the model group had high expression of M1 macrophage markers CD68 and iNOS mRNA, with significant differences (p < 0.0001); the expression of M2 macrophage markers CD163 and Arg-1 mRNA was relatively decreased, but there was a significant difference in CD163 mRNA (P < 0.05); the expressions of CD68, CD163, Arg-1, and CD163 in the traditional Chinese medicine composition group were all higher than those in the control group, with significant differences (p < 0.0001, p < 0.001, P < 0.05); compared with the model group, the expressions of CD68 and iNOS mRNA in the traditional Chinese medicine composition group decreased significantly (p < 0.0001, P < 0.05), while the expressions of CD163 and Arg-1 mRNA increased significantly, with significant differences (p < 0.001), tending to the M2 phenotype. See Table 5.

[0080] Table 5 CD68, iNOS, CD163, and Arg-1 mRNA in the renal tissues of IgA nephropathy rats Results

[0081] Grouping N CD68 iNOS CD163 Arg-1 Control group 3 1.08±0.18 0.89±0.10 1.14±0.19 0.96±0.13 Model group 3 5.03±0.22**** 2.80±0.16**** 1.78±0.18* 1.37±0.16 Traditional Chinese medicine composition group 3 3.08±0.25****#### 1.33±0.16*#### 3.69±0.32****### 2.46±0.24***###

[0082] Note: Compared with the control group, ****p < 0.0001, ***P < 0.001, **P < 0.01; compared with the model group, #p < 0.0001; P < 0.001.

[0083] 5. Detection of TRAF6 and NF-κBp65 mRNA in kidney tissues by q-RTPCR

[0084] The expressions of TRAF6 / NF-κBp65 mRNA in the renal tissues of IgA nephropathy rats showed that: compared with the control group, the expressions of TRAF6 and NF-κBp65 mRNA in the model group increased significantly, with significant differences (p < 0.0001, P < 0.01); the expressions of TRAF6 and NF-κBp65 mRNA in the traditional Chinese medicine composition group were both higher than those in the control group, and there was a significant difference in NF-κBp65 mRNA (P < 0.05); compared with the model group, the expressions of TRAF6 and NF-κBp65 mRNA decreased significantly, with significant differences (p < 0.001, p < 0.05). See Table 6.

[0085] Table 6 TRAF6 and NF-κBp65 mRNA in the renal tissues of IgA nephropathy rats

[0086] Grouping N TRAF6 NF-KBp65 Control group 3 1.09±0.13 1.06±0.17 Model group 3 2.67±0.18**** 2.06±0.21** Traditional Chinese medicine composition group 3 1.40±0.14### 1.57±0.16*#

[0087] Note: Compared with the control group, ****p < 0.0001, **P < 0.01, *P < 0.05; compared with the model group, P < 0.001, ##P < 0.01.

[0088] 6. Detection of TRAF6, p-NF-κB, and NF-κBp65 proteins in renal tissue

[0089] The expression levels of TRAF6 and its downstream NF-κBp65 proteins in renal tissue were detected by Western blotting. The results showed that compared with the control group, the expression levels of TRAF6, p-NF-κBp65, and NF-κBp65 proteins in the model group were higher than those in the control group, with significant differences ((p < 0.0001, p < 0.001, P < 0.05). The content of TRAF6 protein in the traditional Chinese medicine composition group was higher than that in the control group, with a significant difference (p < 0.01). Compared with the model group, the levels of TRAF6, p-NF-κBp65, and NF-κBp65 proteins in the traditional Chinese medicine composition group decreased, with significant differences (p < 0.01, P < 0.05). See Table 7.

[0090] Table 7 Contents of TRAF6, p-NF-κB, and NF-κBp65 proteins in renal tissue of IgA nephropathy rats

[0091] Grouping TRAF6 p-NF-KB NF-KBp65 Control group 3 0.27±0.06 0.29±0.06 0.75±0.09 Model group 3 0.81±0.04**** 0.74±0.12*** 1.01±0.08* Traditional Chinese medicine composition group 3 0.75±0.09**## 0.41±0.03## 0.80±0.04#

[0092] Note: Compared with the control group, ****p < 0.0001, ***P < 0.001, *P < 0.05; compared with the model group, ##P < 0.01, #P < 0.05.

[0093] In animal experimental studies, the traditional Chinese medicine composition of the present invention has a good therapeutic effect on proteinuria, can improve renal function, reduce intestinal sIgA secretion, promote the transformation of renal macrophages into an anti-inflammatory phenotype, reduce the contents of inflammatory cytokines MCP-1, TNF-α, IL-1β, and IL-6 in renal tissue, and inhibit the expression of genes and proteins related to the inflammatory signaling pathway TRAF6 / NF-κBp65.

[0094] Example 2

[0095] Clinical observation of the traditional Chinese medicine composition in the treatment of membranous nephropathy proteinuria

[0096] Sixty patients with primary membranous nephropathy who visited the doctor from March 2021 to March 2024 were selected and divided into a control group and a traditional Chinese medicine composition group in an open-label manner, with 30 patients in each group. There were 18 males and 12 females in the control group; the age was (50 ± 13.41) years; the disease course was (4.17 ± 1.35) years; there were 18 males and 12 females in the traditional Chinese medicine composition group; the age was (49.84 ± 12.47) years; the disease course was (4.08 ± 1.29) years. When comparing the general data of the two groups of patients, the differences were not statistically significant (P > 0.05), and they were comparable.

[0097] 1. Grouping and treatment methods

[0098] General treatment: For all the selected patients, antihypertensive, lipid-lowering, anticoagulant and other treatments were given according to the condition, with the principle of not interfering with the experimental research.

[0099] Control group: In addition to general treatment, immune treatment was the main treatment. According to the pathology and proteinuria situation, immune treatments such as hormones and / or cyclophosphamide, tacrolimus, rituximab, etc. were routinely given. One course of treatment was 3 months, and observation was carried out for 3 months.

[0100] Traditional Chinese medicine composition group: According to the condition, general Western medicine basic treatment was combined with the traditional Chinese medicine composition, or the traditional Chinese medicine composition was added on the basis of applying hormone immunosuppressants (150 mL each time, twice a day, taken separately in the morning and evening). One course of treatment was 3 months, and observation was carried out for 3 months.

[0101] Preparation method of the traditional Chinese medicine composition: The following traditional Chinese medicine raw materials in parts by weight: 30 g of Astragalus membranaceus, 15 g of stir-fried Atractylodes macrocephala, 10 g of Polygonum cuspidatum, 10 g of Alisma orientale, 15 g of Smilax glabra, 10 g of Curcuma longa, 10 g of Pheretima aspergillum, 10 g of Bombyx batryticatus, 15 g of Tripterygium regelii, 15 g of Centella asiatica, 15 g of Bidens pilosa and 15 g of Limonium bicolor were mixed to obtain a mixture.

[0102] Water equivalent to 10 times the volume of the mixture was added to the mixture.

[0103] Heat, and after boiling, change to slow fire and boil until the remaining liquid is 8 times the volume of the mixture. After stopping the fire, cool down, filter, collect the medicinal residues and filtrate, and add water equivalent to 6 times the volume of the medicinal residues to the medicinal residues.

[0104] Repeat the decocting step 3 times, combine all the filtrates, concentrate to 150 mL, and obtain the traditional Chinese medicine stock solution with a crude drug concentration of 1.13 g / mL, that is, the traditional Chinese medicine composition for treating nephrotic proteinuria.

[0105] 2. Observation indicators

[0106] 1) Statistically analyze the TCM syndrome scores of the two groups of patients before and after treatment, including 4 items: fatigue, soreness and coldness of the waist and knees, limb edema, and frequency of nocturia. They were scored 0 point, 2 points, 4 points, and 6 points respectively according to none, mild, moderate, and severe. The higher the score, the more serious the patient's condition.

[0107] 2) 24-hour urine protein index: Compare the 24-hour urine protein quantification before and after treatment in both groups. Observe the occurrence of adverse reactions during the treatment in both groups of patients.

[0108] 3) Efficacy judgment criteria: The judgment criteria were formulated according to the "Guiding Principles for Clinical Research of New Chinese Medicines (Trial Edition)" (2002 edition). The specific criteria are as follows:

[0109] (1) Clinical cure: The clinical symptoms of the patient completely disappear or basically disappear, with basically no discomfort, and the efficacy index is greater than or equal to 90%;

[0110] (2) Marked effect: The clinical symptoms of the patient are significantly improved, and the discomfort is significantly improved, 60% ≤ efficacy index < 90%;

[0111] (3) Effective: The clinical symptoms of the patient are somewhat improved, and the discomfort is improved, 30% ≤ efficacy index < 60%;

[0112] (4) Ineffective: The clinical symptoms of the patient are aggravated or not significantly improved, the discomfort is aggravated or not significantly improved, and the efficacy index < 30%.

[0113] The efficacy index is calculated according to the Nimodipine method, that is: Efficacy index (N) = [(starting score - completion score) / starting score] × 100%.

[0114] 4. Statistical methods

[0115] SPSS 19.0 and Graphpad 8.0 statistical software were used for statistical analysis. Analysis of variance was used for comparison before and after treatment and between groups, and the χ2 test was used for comparison of rates. The results of measurement data are expressed as mean ± standard deviation P < 0.05 indicates that the difference is statistically significant.

[0116] 5. Detection results

[0117] The results of 24-hour urine protein detection showed that there was a significant difference P < 0.05 in the comparison before and after treatment in the control group, and there was a significant difference P < 0.05 in the Chinese medicine composition group before and after treatment. After treatment, there was a significant difference P < 0.05 in the comparison between the Chinese medicine composition group and the control group after treatment. See Table 8.

[0118] For the comparison of clinical efficacy, the clinical effective rate of the control group was 70%, and the clinical effective rate of the Chinese medicine composition group was 86.7%; see Table 9.

[0119] The results of clinical syndrome scores showed that there was a significant difference P < 0.01 in the comparison before and after treatment in the control group, and there was a significant difference P < 0.01 in the Chinese medicine composition group before and after treatment. After treatment, there was a significant difference P < 0.01 in the comparison between the Chinese medicine composition group and the control group after treatment. See Table 10.

[0120] Table 8 Comparison of 24-hour urinary protein between two groups before and after treatment (g / L)

[0121]

[0122] Note: Comparison within each group before and after treatment in the two groups, *P < 0.05; comparison between the two groups after treatment, #*P < 0.05.

[0123] Table 9 Comparison of clinical efficacy between two groups of patients (cases)

[0124] Group N Clinical control Marked effect Effective Ineffective Effective rate % Control group 30 1 8 12 9 70 Traditional Chinese medicine composition group 30 2 15 9 4 86.7

[0125] Table 10 Comparison of clinical syndrome scores between two groups of patients

[0126]

[0127] Note: Comparison within each group before and after treatment in the two groups, **P < 0.01; comparison between the two groups after treatment, ##P < 0.01.

[0128] The results of the clinical study showed that the treatment with the traditional Chinese medicine composition could effectively reduce the urinary protein level of patients, and at the same time effectively relieve the clinical symptoms of patients such as fatigue, soreness and coldness in the waist and knees, limb edema, and frequency of nocturia. The clinical effective rate was 86.7%, which was significantly higher than that of the control group. Therefore, in the clinical treatment of nephrotic proteinuria, the traditional Chinese medicine composition of the present invention has significant clinical efficacy, is superior to the simple western medicine treatment, and provides an effective prescription for the clinical treatment of proteinuria.

[0129] Example 3

[0130] Case 1: Treatment of membranous nephropathy proteinuria with the traditional Chinese medicine composition prepared in Example 2

[0131] Patient Jian Mou, female, 45 years old, with the main complaint of "facial and bilateral lower limb edema for more than 1 year". Western medicine diagnosis: membranous nephropathy. She had been treated with hormones + tacrolimus, cyclophosphamide pulse, rituximab, etc., but the curative effect was poor. On March 1, 2023, the total protein in liver function was 38.8 g / L, albumin was 19.2 g / L, and the 24-hour urinary protein quantification was 11.85 g / L. She came to see a traditional Chinese medicine doctor and started traditional Chinese medicine treatment. The patient took the traditional Chinese medicine composition of the present invention for 18 months. On September 5, 2024, the albumin in liver function rose to normal, and the 24-hour urinary protein quantification was detected to be 0.22 g / L. Figure 1 It is the graph of the liver function test results of Jian Mou; Figure 2 It is the graph of the 24-hour urinary protein quantification results of Jian Mou. It can be seen that after treatment with the traditional Chinese medicine composition, the patient's edema disappeared, the hypoproteinemia improved, and the 24-hour urinary protein quantification was close to the normal level, with significant clinical efficacy. Moreover, the price of the traditional Chinese medicine composition of the present invention is lower than that of drugs such as tacrolimus and rituximab, the curative effect is reliable, and it has long-term economic and social benefits, which can reduce the economic burden of patients.

[0132] Case 2: Treating IgA Nephropathy Proteinuria with the Traditional Chinese Medicine Composition Prepared in Example 2

[0133] Patient Wang, female, 52 years old, with "positive urine protein for 3 years". Diagnosed by renal biopsy: IgA nephropathy. Visited on February 7, 2024, with a 24-hour urine protein quantification of 2.2 g / L. Treatment with the traditional Chinese medicine composition was started. On May 15, 2024, the 24-hour urine protein quantification was 0.67 g / L; on July 12, 2024, the 24-hour urine protein quantification was 0.56 g / L; on October 30, 2024, the 24-hour urine protein quantification was 0.38 g / L, and on February 7, 2025, the 24-hour urine protein quantification was 0.18 g / L. Figure 3 It is a graph of the 24-hour urine protein quantification test results for Wang. After treatment, the patient's proteinuria decreased significantly and approached the normal level.

[0134] Case 3: Treating Chronic Nephritis Proteinuria with the Traditional Chinese Medicine Composition Prepared in Example 2

[0135] Patient Yang, male, 31 years old, with "mild swelling of both lower extremities for 3 months". Diagnosis: Chronic nephritis. On June 4, 2022, the 24-hour urine protein quantification was detected as 1.58 g / L. Traditional Chinese medicine treatment was started, and the traditional Chinese medicine composition was given. One course of treatment was 2 weeks. On September 4, 2022, the 24-hour urine protein quantification was 1.13 g / L. The patient had a cold 1 week before the examination on December 12, 2022, and the 24-hour urine protein quantification was 1.72 g / L. On February 27, 2023, the 24-hour urine protein quantification was 0.53 g / L; on April 8, 2023, the 24-hour urine protein quantification was 0.22 g / L; on June 17, 2023, the 24-hour urine protein quantification was 0.12 g / L, and the patient was clinically cured. (Reference value: 0 - 0.15 g / L). Figure 4 It is a graph of the 24-hour urine protein quantification test results for Yang.

[0136] Case 4: Treating Diabetic Nephropathy Proteinuria with the Traditional Chinese Medicine Composition Prepared in Example 2

[0137] Patient Wang, male, 73 years old, with the main complaint of "high blood sugar for 23 years and swelling of both lower extremities for 6 years". Diagnosis: Type 2 diabetic nephropathy. On October 26, 2024, the 24-hour urine protein quantification was detected as 4.94 g / L. Treatment with the traditional Chinese medicine composition was started. On November 4, 2024, the 24-hour urine protein quantification was 2.88 g / L; on December 10, 2024, the 24-hour urine protein quantification was 2.69 g / L; on February 21, 2025, the 24-hour urine protein quantification was 1.24 g / L; on April 21, 2025, the 24-hour urine protein quantification was 1.10 g / L. Figure 5This is the graph of the 24-hour urinary protein quantification test result of Wang. In the stage of massive proteinuria in diabetic nephropathy, clinical treatment is very difficult. The results show that after treatment with the traditional Chinese medicine composition of the present invention, the patient's edema is significantly reduced, and the urinary protein quantification is significantly decreased, which proves that the traditional Chinese medicine composition of the present invention can effectively reduce proteinuria in patients with diabetic nephropathy and provides an effective therapeutic drug for the treatment of diabetic nephropathy.

[0138] Case 5: Treatment of nephrotic syndrome proteinuria with the traditional Chinese medicine composition prepared in Example 2

[0139] Patient Li, male, 71 years old, with the main complaint of "facial and bilateral lower extremity edema for more than 2 months". Diagnosis: Nephrotic syndrome. Presenting with edema and oliguria, with a daily urine volume of 800 - 1000 ml. On October 15, 2024, the 24-hour urinary protein quantification was 6.51 g / L, and the patient was given treatment with the traditional Chinese medicine composition. On November 22, 2024, the 24-hour urinary protein quantification was 4.25 g / L; on December 25, 2024, the 24-hour urinary protein quantification was 2.51 g / L; on January 28, 2025, the 24-hour urinary protein quantification was 1.89 g / L; on February 27, 2025, the 24-hour urinary protein quantification was 1.35 g / L; on April 15, 2025, the 24-hour urinary protein quantification was 0.96 g / L. Figure 6 This is the graph of the 24-hour urinary protein quantification test result of Li. After treatment, the patient's edema and oliguria improved, and the urinary protein quantification decreased significantly. It shows that the traditional Chinese medicine formula of the present invention has the effects of diuresis, increasing plasma albumin, and reducing proteinuria. During the medication process, the patient did not experience gastrointestinal discomfort, had good tolerance, and was safe and effective.

[0140] The above are only a few examples among many cases of treating proteinuria with the traditional Chinese medicine composition of the present invention, all of which have 24-hour urinary protein test data. As the medication time prolongs, the decrease in proteinuria is more obvious, indicating that the curative effect of traditional Chinese medicine is more significant. The above cases prove that the traditional Chinese medicine composition of the present invention can treat various chronic kidney diseases mainly manifested by proteinuria, can reduce edema, regulate the gastrointestinal tract, and increase the plasma albumin level, especially has significant curative effects on membranous nephropathy, IgA nephropathy, diabetic nephropathy, nephrotic syndrome, chronic glomerulonephritis, etc., and the effect is more significant as the medication time prolongs. No side effects such as liver and kidney damage were found during the treatment with the traditional Chinese medicine composition of the present invention.

[0141] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, not all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition, characterized in that, The raw materials for preparation include the following parts by weight: 30 - 60 parts of Astragalus membranaceus, 10 - 20 parts of stir-fried Atractylodes macrocephala, 8 - 16 parts of Polygonum cuspidatum, 8 - 16 parts of Alisma orientale, 10 - 20 parts of Smilax glabra, 8 - 16 parts of Curcuma longa, 8 - 16 parts of Pheretima aspergillum, 8 - 16 parts of Bombyx batryticatus, 10 - 20 parts of Tripterygium regelii, 10 - 20 parts of Centella asiatica, 10 - 20 parts of Bidens pilosa, and 10 - 20 parts of Limonium bicolor.

2. The traditional Chinese medicine composition according to claim 1, wherein The raw materials for preparation include the following parts by weight: 30 parts of Astragalus membranaceus, 15 parts of stir-fried Atractylodes macrocephala, 12 parts of Polygonum cuspidatum, 12 parts of Alisma orientale, 15 parts of Smilax glabra, 12 parts of Curcuma longa, 12 parts of Pheretima aspergillum, 12 parts of Bombyx batryticatus, 15 parts of Tripterygium regelii, 15 parts of Centella asiatica, 15 parts of Bidens pilosa, and 15 parts of Limonium bicolor.

3. The preparation method of the traditional Chinese medicine composition according to claim 1 or 2, comprising the following steps: Mix the raw materials with water, soak, decoct, and concentrate to obtain the traditional Chinese medicine composition.

4. The preparation method according to claim 3, wherein The soaking time is 1.5 - 2.5 h; the number of decoction times is 2 - 3 times; after concentration, the crude drug concentration is 1 - 2.5 g / mL.

5. A traditional Chinese medicine preparation made from the traditional Chinese medicine composition according to claim 1 or 2.

6. The traditional Chinese medicine preparation according to claim 5, characterized in that, The dosage form of the traditional Chinese medicine preparation includes capsules, granules, medicinal plasters, or cut crude drugs.

7. The application of the traditional Chinese medicine composition according to claim 1 or 2, or the traditional Chinese medicine composition prepared by the preparation method according to claim 3 or 4, or the traditional Chinese medicine preparation according to claim 5 or 6 in the preparation of a drug for treating chronic kidney disease.

8. The application according to claim 7, wherein The chronic kidney disease includes any one or more of IgA nephropathy, membranous nephropathy, nephrotic syndrome, diabetic nephropathy, and chronic glomerulonephritis.

9. The application of the traditional Chinese medicine composition according to claim 1 or 2, or the traditional Chinese medicine composition prepared by the preparation method according to claim 3 or 4, or the traditional Chinese medicine preparation according to claim 5 or 6 in the preparation of a drug for treating renal proteinuria.

10. The application of the traditional Chinese medicine composition according to claim 1 or 2, or the traditional Chinese medicine composition prepared by the preparation method according to claim 3 or 4, or the traditional Chinese medicine preparation according to claim 5 or 6 in the preparation of a drug having any one or more of the following effects: ① Improving renal function; ② Reducing the level of renal tissue inflammatory factors; ③ Promoting the transformation of renal macrophages into anti-inflammatory phenotypes; ④ Inhibiting the gene and protein expression of molecules related to the inflammatory signaling pathway TRAF6 / NF-κBp65.

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