A traditional Chinese medicine composition for treating chronic kidney disease, its preparation method and uses

Abstract

The present invention provides a traditional Chinese medicine composition for treating chronic kidney disease, which is prepared from the following raw materials in the following weight ratios: 4-6 parts of cimicifuga, 24-36 parts of rhubarb, 8-12 parts of polygonum cuspidatum, 12-18 parts of scutellaria baicalensis, 8-12 parts of artemisia capillaris, 4-6 parts of wax gourd peel, 4-6 parts of motherwort, 4-6 parts of leech, 4-6 parts of earthworm, 4-6 parts of safflower, and 4-6 parts of millettia reticulata. The present invention also provides a preparation method and use of the traditional Chinese medicine composition. The action site of the drug of the present invention focuses on the intestine, and enema directly acts on the colon, which can accelerate intestinal kinetics, improve intestinal barrier function, reduce the production and absorption of enterogenous uremic toxins such as indoxyl sulfate, reduce renal fibrosis, relieve renal fibrosis, and has a significant correlation with regulating TLR4 / MyD88 / NF-κB inflammatory response and systemic inflammation, providing a new option for the treatment of chronic kidney disease.

Description

Technical Field

[0001] The present invention relates to a traditional Chinese medicine composition for treating chronic kidney disease. Background Art

[0002] Chronic kidney disease (CKD) refers to chronic kidney structural and functional disorders caused by various reasons (history of kidney damage greater than 3 months), including pathological damage with normal and abnormal renal GFR, abnormal blood or urine components, and abnormal imaging examinations, or unexplained decline in GFR (<60 ml / min·1.73m 2 ) for more than 3 months, which is CKD. Epidemiological investigations show that the prevalence of chronic kidney disease is very high globally, and the incidence rates in various countries range from 5% to 13%. Due to the long course of chronic kidney disease, long-term observation and treatment are required. If it progresses to end-stage renal disease and enters renal replacement therapy, the daily lives of patients and their families will be greatly affected, and it will also have a huge impact on social public health care. How to achieve kidney protection for chronic kidney disease patients has always been the focus of treating this disease.

[0003] In the middle and late stages of chronic kidney disease, metabolic wastes cannot be excreted through the intestine and kidney and accumulate in large amounts in the body, and may further enter the intestinal wall through blood vessels. At the same time, the increase in uremic toxins in the blood after renal failure also enters the intestinal wall, promoting fundamental changes in the survival environment of intestinal epithelium and intestinal flora, resulting in an increase in the number of pathogenic bacteria and a decrease in beneficial bacteria, directly or indirectly promoting the entry of toxins into intestinal epithelial cells, causing serious damage to the structure and function of the intestinal barrier, and greatly increasing the permeability of the intestinal epithelium, which in turn leads to the invasion of intestinal toxins into the blood and accelerates the progression of CKD. It can be seen that the intestine and the kidney are closely related. Physiologically, they form the "gut-kidney axis" through two pathways of metabolic dependence and immunity. Pathologically, when the intestinal barrier is damaged, it will promote the occurrence and development of chronic kidney disease. In terms of treatment, studies have found that traditional Chinese medicine can not only reduce intestinal inflammation, repair the colonic mucosal barrier and restore the balance of intestinal flora, but also play an important role in the prevention and management of sepsis-related kidney injury by reducing the levels of serum creatinine, blood urea nitrogen, TNF-α, NF-κB, etc. and increasing the activities of antioxidant enzymes and the level of PPARγ [YE N, ZHAO P, AYUE S, et al. Folic acid-modified lactoferrin nanoparticles coated witha laminarin layer loaded curcumin with dual-targeting for ulcerative colitistreatment[J]. Int J Biol Macromol, 2023, 232(123229, BALKRISHNA A, SINHA S, KUMARA, et al. Sepsis-mediated renal dysfunction:Pathophysiology, biomarkers and roleof phytoconstituents in its management[J]. Biomed Pharmacother, 2023, 165(115183].

[0004] As a research hotspot in recent years, the gut-kidney axis indicates that the intestine is an important target for the treatment of chronic kidney disease. The intestinal mucosal immune system is one of the largest immune compartments in the human body, and any disruption of the microbiome may lead to imbalance of the human immune system. Therefore, it is a new direction worth exploring to use modern medical methods to explore the mechanism of intestinal microecological balance and the interaction between intestinal mucosal barrier function and CKD through in vivo animal models and in vitro cell models. Lipopolysaccharide (LPS) is an inflammatory marker involved in the pathogenesis of CKD. Intestinal microbiota dysbiosis inhibits the expression of tight junction proteins, leading to increased intestinal permeability and translocation of LPS derived from Gram-negative bacteria into the blood, which may be related to metabolic inflammation and the progression of CKD. Studies have shown that the concentration of indoxyl sulfate (IS) in healthy people is 0.10 - 2.39 μM, while the total IS concentration in CKD patients exceeds 500 μM. As renal function declines, IS can cause kidney damage by generating reactive oxygen species (ROS), damaging the antioxidant system, increasing inflammatory responses, inducing fibrosis, and reducing protective proteins.

[0005] At present, the intestinal administration method of traditional Chinese medicine in the treatment of chronic kidney disease has been reported in relevant literature. For example, Yi Xiaofeng, et al., Multiple Traditional Chinese Medicine Treatments for Chronic Renal Failure, China Traditional Chinese Medicine News / April 14, 2010 / Edition 005, Retention enema with traditional Chinese medicine can avoid the direct stimulation of oral laxatives on the digestive tract through the intestinal administration route, and can also increase the excretion of urea nitrogen, creatinine, uric acid, potassium, phosphorus, etc. The so-called "the six fu-organs function by thoroughfare" and "when the pathogen is removed, the healthy qi will be restored", and it has an ideal clinical effect on early and middle-stage chronic renal failure, but the specific traditional Chinese medicine prescription is not disclosed. The Professional Committee of Nephrology Diseases of the Chinese Association of Integrated Traditional Chinese and Western Medicine, Expert Consensus on the Rational Use of Shenkang Preparations (Shenkang Injection, Shenkang Suppository) in the Treatment of Chronic Kidney Disease, Chinese Journal of Integrated Traditional and Western Medicine, Vol. 39, No. 7, July 2019, disclosed that Shenkang Suppository prepared from four traditional Chinese medicines, namely rhubarb, salvia miltiorrhiza, safflower, and astragalus membranaceus, is used for chronic renal failure with the syndrome of dampness turbidity and blood stasis, and has the effects of reducing adverse qi and discharging turbidity, replenishing qi and activating blood circulation. The main action site is the kidney. Based on the principle of syndrome differentiation and treatment of traditional Chinese medicine, according to different clinical manifestations and physical characteristics, chronic kidney disease has different syndromes, such as: syndrome of spleen-kidney qi deficiency, syndrome of spleen-kidney yang deficiency, syndrome of qi-yin deficiency, syndrome of liver-kidney yin deficiency, syndrome of yin-yang deficiency, syndrome of dampness turbidity, syndrome of dampness-heat, syndrome of fluid retention, syndrome of blood stasis, syndrome of turbid toxin, etc. The traditional Chinese medicine formulas reported so far cannot meet the clinical needs. Summary of the Invention

[0006] The technical solution of the present invention provides a new traditional Chinese medicine composition for the treatment of chronic kidney disease, and the present invention also provides a preparation method and use of the traditional Chinese medicine composition.

[0007] The present invention provides a traditional Chinese medicine composition for the treatment of chronic kidney disease, which is prepared from the following raw materials in the following weight ratios:

[0008] 4 - 6 parts of Cimicifuga foetida, 24 - 36 parts of Rheum palmatum, 8 - 12 parts of Polygonum cuspidatum, 12 - 18 parts of Scutellaria baicalensis, 8 - 12 parts of Artemisia capillaris, 4 - 6 parts of wax gourd peel, 4 - 6 parts of Leonurus japonicus, 4 - 6 parts of Hirudo, 4 - 6 parts of Pheretima aspergillum, 4 - 6 parts of Carthamus tinctorius, 4 - 6 parts of Spatholobus suberectus

[0009] Further preferably, it is prepared from the raw medicinal materials in the following weight ratios:

[0010] 5 parts of Cimicifuga foetida, 30 parts of Rheum palmatum, 10 parts of Polygonum cuspidatum, 15 parts of Scutellaria baicalensis, 10 parts of Artemisia capillaris, 5 parts of wax gourd peel, 5 parts of Leonurus japonicus, 5 parts of Hirudo, 5 parts of Pheretima aspergillum, 5 parts of Carthamus tinctorius, 5 parts of Spatholobus suberectus

[0011] The traditional Chinese medicine composition of the present invention takes the raw medicinal powder of the above - mentioned raw medicinal materials, water or organic solvent extract as active ingredients, and is prepared into a pharmaceutically common intestinal administration preparation by adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0012] Among them, the intestinal administration preparation includes enemas, suppositories, and intestinal foams.

[0013] The present invention also provides a preparation method of the above - mentioned traditional Chinese medicine composition, which includes the following steps:

[0014] a. Weigh the raw materials in each weight ratio and crush them;

[0015] b. Decoct with water, concentrate the decoction, and add pharmaceutically acceptable excipients or auxiliary ingredients to prepare a pharmaceutically common intestinal administration preparation.

[0016] The present invention also provides the use of the above - mentioned traditional Chinese medicine composition in the preparation of a drug for intestinal administration for treating chronic kidney disease.

[0017] Among them, the drug is a drug that intervenes in the gut - kidney axis and TLR4 / MyD88 / NF - κB pathway, reduces creatinine, proteinuria, endotoxin, and inflammatory factors, inhibits renal fibrosis, and delays the decline of renal function in chronic kidney disease.

[0018] The drug is used to treat chronic kidney disease with water qi syndrome or blood stasis syndrome.

[0019] The present invention also provides the use of the above - mentioned traditional Chinese medicine composition in the preparation of a drug for intestinal administration with the functions of removing turbid dampness and promoting blood circulation and diuresis.

[0020] In "Plain Questions of Huangdi Neijing", it is stated that "The large intestine is an official in charge of transmission and transformation, where changes occur." The raw materials of the medicine in the present invention are based on the physiological process of the intestine transmitting turbid qi, and the method of enema is adopted to improve the disease progression of chronic kidney disease. After the onset of chronic kidney disease, the warming and steaming function of the kidney weakens. Insufficient yang qi leads to metabolic disorders and damage to the qi movement of the five zang-organs. The function of the intestine in secreting and transmitting turbid qi declines, and external and internal pathogenic factors such as dampness turbidity and stasis cannot be discharged, further affecting the qi and blood circulation and normal functions of the whole body's zang-fu organs, resulting in complications such as vomiting, fatigue, poor appetite, palpitation, and edema, increasing the burden on the kidneys. Therefore, the method of enema for secreting turbid qi is used to discharge turbid qi from the intestine, promote the intestine to secrete and excrete toxins such as dampness turbidity and stasis, and improve the physiological environment and function of the kidneys.

[0021] In the raw material formula of the present invention, rhubarb is the monarch drug, with its functions of promoting blood circulation, purging, detoxifying, and promoting diuresis; polygonum cuspidatum and cimicifuga foetida are the minister drugs. Polygonum cuspidatum is bitter in taste and cold in nature, with the effects of promoting diuresis, detoxifying, and promoting blood circulation, assisting rhubarb to strengthen the power of purging the bowels and discharging turbid qi. Cimicifuga foetida ascends the clear qi to descend the turbid qi, and the ascending and descending complement each other to enhance the purging power of rhubarb; Scutellaria baicalensis, artemisia capillaris, and wax gourd peel are used as assistant drugs. Among them, Scutellaria baicalensis clears heat and dries dampness, artemisia capillaris clears heat and promotes diuresis, and wax gourd peel promotes diuresis and percolates dampness, jointly exerting the effect of detoxifying and discharging turbid qi; Leonurus japonicus, hirudo, pheretima, carthamus tinctorius, and millettia reticulata are used as assistant and guiding drugs. Among them, Leonurus japonicus promotes diuresis and activates blood circulation, hirudo breaks blood stasis, pheretima dredges the channels and collaterals, carthamus tinctorius activates blood circulation and dredges the channels, and millettia reticulata activates blood circulation and nourishes blood, removing stasis and generating new blood, jointly exerting the effects of secreting turbid qi, removing dampness, activating blood circulation, and promoting diuresis, and improving the kidney function.

[0022] The raw materials of the medicine in the present invention are composed of raw medicinal materials with the effects of expelling wind (cimicifuga foetida), discharging turbid qi (rhubarb, polygonum cuspidatum), removing dampness (Scutellaria baicalensis, artemisia capillaris, etc.), and dredging collaterals (hirudo, pheretima, etc.). The action site focuses on the intestine. The traditional Chinese medicine enema directly acts on the colon, which can accelerate intestinal kinetics, improve intestinal barrier function, reduce the production and absorption of enterogenous uremic toxins such as indoxyl sulfate, reduce renal fibrosis, relieve renal fibrosis, and has a significant correlation with regulating the TLR4 / MyD88 / NF-κB inflammatory response and systemic inflammation, providing a new option for the treatment of chronic kidney disease. Brief Description of the Drawings

[0023] Figure 1 The repair effect of the enema solution for secreting turbid qi on the renal pathological injury of CKD rats;

[0024] Figure 2 The effects of the enema solution for secreting turbid qi on BUN, SCR, and 24-hour urinary protein in the CKD rat model;

[0025] Figure 3 The repair of intestinal pathological injury by the enema solution for secreting turbid qi on intestinal junction proteins (ZO-1, Occludin-1, and Claudin-1) in the CKD rat model;

[0026] Figure 4Inhibitory effect of Bizhuo Enema on intestinal endotoxin (IS, LPS) and inflammatory factors (IL-6, TNF-α) in CKD rat model Detailed implementation mode

[0027] Example 1 Preparation of the drug of the present invention (hereinafter referred to as Bizhuo Enema (MZGCY))

[0028] Prescription is as follows: Cimicifuga foetida 5g, Rheum palmatum 30g, Polygonum cuspidatum 10g, Scutellaria baicalensis 15g, Artemisia capillaris 10g, Benincasa hispida peel 5g, Leonurus japonicus 5g, Hirudo 5g, Pheretima 5g, Carthamus tinctorius 5g, Spatholobus suberectus 5g.

[0029] The preparation process is as follows:

[0030] (1) Crushing: Put the above-mentioned Chinese herbal pieces into a crusher respectively and crush them into coarse powder for easy decoction.

[0031] (2) Decoction: For the first time, decoct with 1000 ml of water and all the drugs until about 400 ml, and take the supernatant. For the second time, add 600 ml of water to the drug residues and decoct until about 200 ml, and take the supernatant. Mix the supernatants obtained twice to get the decoction.

[0032] (3) Concentration: Let it stand, filter to remove impurities, then take the supernatant for concentration, add distilled water to 100 ml, sterilize for 30 minutes, and store in sealed and refrigerated condition. The drug content is 1 g / ml.

[0033] Example 2 Preparation of the drug of the present invention

[0034] Prescription: Cimicifuga foetida 4g, Rheum palmatum 24g, Polygonum cuspidatum 8g, Scutellaria baicalensis 12g, Artemisia capillaris 8g, Benincasa hispida peel 4g, Leonurus japonicus 4g, Hirudo 4g, Pheretima 4g, Carthamus tinctorius 4g, Spatholobus suberectus 4g;

[0035] Prepared according to the preparation method of Example 1.

[0036] Example 3 Preparation of the drug of the present invention

[0037] Prescription: Cimicifuga foetida 6g, Rheum palmatum 36g, Polygonum cuspidatum 12g, Scutellaria baicalensis 18g, Artemisia capillaris 12g, Benincasa hispida peel 6g, Leonurus japonicus 6g, Hirudo 6g, Pheretima 6g, Carthamus tinctorius 6g, Spatholobus suberectus 6g;

[0038] Prepared according to the preparation method of Example 1.

[0039] The beneficial effects of the present invention are demonstrated by the following pharmacodynamic experiments.

[0040] Test Example 1 Pharmacodynamic test of the drug of the present invention

[0041] 1. Experimental materials

[0042] 1.1 Experimental animals

[0043] Male Sprague-Dawley rats (6 - 8 weeks old, 220 - 240 g) were obtained from SPF Biotechnology Co., Ltd. (Beijing, China).

[0044] 1.2 Experimental drugs

[0045] The traditional Chinese medicine prescription of Mizhuo Enema Solution (MZGCY) is as follows: Cimicifuga foetida 5 g, Rheum palmatum 30 g, Polygonum cuspidatum 10 g, Scutellaria baicalensis 15 g, Artemisia capillaris 10 g, Benincasa hispida peel 5 g, Leonurus japonicus 5 g, Hirudo 5 g, Pheretima 5 g, Carthamus tinctorius 5 g, Spatholobus suberectus 5 g.

[0046] Control group: Rhubarb group, the prescription is as follows: Rheum palmatum 100 g.

[0047] All traditional Chinese medicines were purchased from the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine.

[0048] 2. Experimental methods

[0049] 2.1 Animal feeding

[0050] The rats were housed in the Animal Center of Chengdu University of Traditional Chinese Medicine, with free access to food and water, and under closed management. They were adaptively fed for one week.

[0051] 2.2 Animal grouping

[0052] All rats were numbered with ear tags, and then random numbers were generated using SPSS software and divided into five groups: blank group, model group, half-dose compound group (enema with 1 / 2 Mizhuo Enema Solution), compound group (enema with full-dose Mizhuo Enema Solution), and rhubarb group (the adult dosage is 1.67 g / kg / d, and the equivalent dosage for rats is 10.37 g / kg / d), with 6 rats in each group.

[0053] 2.3 Experimental model establishment

[0054] After the experiment started, except for the blank group, the other 4 groups were given adenine to establish a chronic kidney disease animal model according to the Yokozawa method.

[0055] In the morning, rats were given intragastric administration of 2.5% adenine suspension (200 mg / d / kg). At the end of the fourth week, two rats were randomly selected from each group, and blood was collected from the medial canthus of the eye to measure serum creatinine and blood urea nitrogen to evaluate the model establishment. In the fifth week, 2.5% adenine suspension (200 mg / d / kg) was given intragastric administration every other day to maintain the disease progression, and adenine intragastric administration was stopped at the 7th week. During this period, the blank group was given an equal volume of distilled water for intragastric administration.

[0056] 2.4 Experimental drug administration

[0057] After the experiment started, the rats in each group were given the corresponding drug treatment in the afternoon. According to the equivalent dose ratio between humans and animals, the adult dosage is 1.67 g / kg / d. The dosage for rats was calculated based on the adult dosage. The total daily dosage for rats was 10.37 g / kg / d (calculated based on the original drug amount). A straight-tip gavage needle was inserted about 8 cm into the rectum of the rats, and the anus was pinched and the gavage needle was fixed. The drug was slowly injected, and the anus was pinched for about 2 minutes to achieve the effect of retention enema.

[0058] 2.5 Specimen collection

[0059] On the last day of the eighth week when the experiment ended, all rats were fasted for 12 hours and anesthetized by intraperitoneal injection of 3% pentobarbital sodium at a dose of 0.2 ml / 100 g. After anesthesia, blood was taken, and bilateral kidneys and intestinal specimens were removed to make pathological sections, which were stained with HE and Masson respectively.

[0060] 2.6 Index detection

[0061] Peripheral blood was collected from the rats, and the levels of intestinal endotoxin indole (including sulfate (IS) and lipopolysaccharide (LPS)) and pro-inflammatory cytokines (TNF-α, IL-6) in each group of rats were measured using an enzyme-linked immunosorbent assay (ELISA) kit. Blood urea nitrogen was detected using an automatic biochemical analyzer; the urine protein concentration was detected using an automatic analyzer with the bicinchoninic acid colorimetric method. TNF-α, IL-6: Multi Science (Lianke) Biotech Co., Ltd

[0062] 2.6.1 Morphological changes under light microscopy

[0063] To examine the intestinal and renal pathological changes in CKD rats, we used hematoxylin and eosin (H&E) staining and then observed using an optical microscope (CX33, Olympus, Tokyo, Japan). At the same time, to observe the improvement degree of renal fibrosis in CKD model rats, we used Masson's trichrome staining (Masson) and periodic acid-Schiff staining (PAS) according to the standard protocol.

[0064] 2.6.2 Immunohistochemical staining

[0065] Paraffin sections of ileum and colon tissues were washed with PBS and fixed with 4% paraformaldehyde. Immunohistochemical staining was performed to evaluate the expression of occludin, claudin-1, ZO-1, and DAPI (nuclear staining). After sealing, images were observed and captured with a fluorescence microscope (Nikon Eclipse C1, Japan), and the fluorescence intensity was measured using ImageJ software. ZO-1 antibody, claudin-1 antibody, and occludin antibody: Protein-tech Group, Inc. (Wuhan, China)

[0066] 2.6.3 Immunofluorescence staining

[0067] To confirm the role of traditional Chinese medicine in regulating the intestine-kidney axis through the NF-κB signaling pathway to improve CKD, we used immunofluorescence staining for relevant verification. Paraffin sections of ileum, colon, and kidney were dewaxed, hydrated, and antigenically repaired. After blocking with 10% normal goat serum, the sections were incubated overnight at 4°C in the primary antibody. The nuclei were stained with DAPI. Then, they were washed with PBS, autofluorescence quencher was added dropwise, and they were blocked with an anti-fluorescence quenching blocker. After sealing, the sections were observed under a fluorescence microscope (Nikon Eclipse C1, Japan) and corresponding images were obtained. NF-κB antibody, TLR4 antibody, MyD88 antibody, Inc. (Wuhan, China)

[0068] 3. Statistical analysis

[0069] SPSS 22 software was used for statistical analysis of the data. For continuous variables, the mean ± standard deviation was used. If the data did not conform to a normal distribution, non-parametric tests were used; when the data conformed to a normal distribution, one-way ANOVA was used for between-group comparisons. When the variances were homogeneous, the LSD method was used for two-group comparisons, and when the variances were heterogeneous, the Dunnett's T3 method was used. A P value < 0.05 was considered statistically significant.

[0070] 4. Experimental results

[0071] 4.1 Pathological morphological changes of the kidneys in each group of rats (see Figure 1 )

[0072] In the model group, adenine metabolite crystal deposits were found in the kidneys, with a large number of inflammatory cell infiltrations, glomerular structural disorders, renal interstitial fibrosis, tubular cystic dilatation, a large number of lumen occlusions, and obvious mesangial hyperplasia, which were consistent with the renal pathological changes in adenine model rats. After treatment with rhubarb and Bizhuo enema solution, glomerular hypertrophy, mesangial matrix expansion, and tubulointerstitial damage were partially improved. Sections stained with Masson trichrome showed that renal fibrosis was improved after treatment with traditional Chinese medicine, and the PAS score of extracellular matrix (ECM) accumulation also decreased. Moreover, the improvement effect of the compound group of Bizhuo enema solution was better than that of the rhubarb group. The above analysis method shows that Bizhuo enema solution has the ability to reduce renal tissue damage in adenine model rats.

[0073] In terms of renal function detection (see Figure 2 ), compared with the blank group, the levels of BUN, SCR, and 24-hour urinary protein in the model group were significantly increased (P < 0.01). Compared with the model group, the levels of BUN, SCR, and 24-hour urinary protein in the half-dose compound group decreased, and there were significant statistical differences in SCR and 24-hour urinary protein (P < 0.01). Compared with the model group, the levels of BUN, SCR, and 24-hour urinary protein in the full-dose compound group decreased (P < 0.05). Compared with the rhubarb group, the levels of BUN, SCR, and 24-hour urinary protein in the compound group of Bizhuo enema solution decreased (P < 0.05), and there were significant statistical differences in BUN, SCR, and 24-hour urinary protein (P < 0.05).

[0074] 4.2 Morphological changes in the ileum and colon of rats in each group are shown in Figure 3

[0075] The intestinal barrier is crucial for maintaining intestinal function. Therefore, we detected the expression of intestinal barrier indicators in rats in each group. The results of H&E staining showed that compared with the model group, treatment with rhubarb and Bizhuo enema solution reduced intestinal damage in model rats. The results of immunohistochemical staining showed that in the control group, ZO-1, Occludin-1, and claudin-1 proteins were evenly distributed at the top of intestinal epithelial cells, showing a honeycomb or dot-like pattern. However, these proteins were significantly reduced in the intestines of the model group. Compared with the model group, the protein levels of ZO-1, Occludin-1, and claudin-1 in the intestines of the compound group were significantly restored, and the compound group of Bizhuo enema solution was better than the rhubarb group. These results indicate that Bizhuo enema solution has a reparative effect on the intestinal barrier of model rats.

[0076] Compared with the blank group (see Figure 4) The levels of IS, LPS, IL-6, and TNF-α in the model group were significantly increased (P < 0.01). Compared with the model group, the levels of IS, LPS, IL-6, and TNF-α in the half-dose compound group all decreased. There was a significant statistical difference in terms of IS (P < 0.01), and a statistical difference in terms of IL-6 (P < 0.05). Compared with the model group, the levels of IS, LPS, IL-6, and TNF-α in the full-dose compound group all decreased. Among them, there were significant statistical differences in terms of IS, IL-6, and TNF-α (P < 0.01). Compared with the rhubarb group, the levels of LPS, IL-6, and TNF-α in the full-dose compound group all decreased (P < 0.05), and there was no statistical difference in terms of IS between the two groups (P > 0.05). These results indicate that Bizhuo Enema Solution has an inhibitory effect on intestinal endotoxin and inflammatory factors in model rats.

[0077] The activation of the NF-κB signaling pathway is the key to the pathogenesis of CKD, which can promote the inflammatory response, regulate cell apoptosis, and vascular remodeling. Our preliminary study shows that the therapeutic effect of Bizhuo Enema Solution on adenine-induced CKD rats is related to the inhibition of the NF-κB signaling pathway. Therefore, to further study the mechanism of Bizhuo Enema Solution in treating CKD through the gut-kidney axis, we evaluated the expression levels of proteins related to the NFκB signaling pathway by immunofluorescence (IF). As shown in the figure, compared with the control group, the fluorescence intensities of TLR4, MyD88, and NF-κB proteins in the ileum and colon tissues of the model group increased. In addition, the fluorescence intensities of TLR4, MyD88, and NF-κB proteins in the compound group of Bizhuo Enema Solution were significantly lower than those in the model group and the rhubarb group.

[0078] 5. Discussion

[0079] According to the theory of traditional Chinese medicine formula compatibility, the traditional Chinese medicine composition of the present invention has the effects of expelling wind (Cimicifuga foetida), purging turbidity (Rheum palmatum, Polygonum cuspidatum), removing dampness (Scutellaria baicalensis, Artemisia capillaris, etc.), and dredging collaterals (Hirudo nipponica, Pheretima aspergillum, etc.). The exertion of its curative effect is closely related to its active ingredient substances. The colon is an important organ for producing uremic toxins. Traditional Chinese medicine enema acts directly on the colon, which can accelerate intestinal kinetics, improve intestinal barrier function, regulate intestinal flora, reduce the production and absorption of enterogenic uremic toxins such as indoxyl sulfate, reduce renal fibrosis, and delay the progression of the kidneys. By enema with Bizhuo Enema Solution of the present invention, the intestinal barrier can be improved, the intestinal flora imbalance can be regulated, the systemic inflammation can be inhibited, and renal fibrosis can be alleviated. The exertion of this effect has a significant correlation with intervening in the alteration of intestinal flora and intestinal barrier markers, TLR4 / MyD88 / NF-κB inflammatory response, and systemic inflammation.

[0080] In summary, TLR4 / MyD88 / NF-κB plays an important role in the occurrence and development of CKD. After the intervention of Bizhuo Enema Solution in CKD rats, the levels of IL-6, TNF-α, TLR4, MyD88, and NF-κB in the intestine of rats were significantly decreased, thereby repairing the intestinal barrier, reducing renal tissue damage, and the curative effect was significantly better than that of the rhubarb group. It is suggested that Bizhuo Enema Solution can improve renal injury in CKD rats, which may be related to its down-regulation of the expression of TLR4 / MyD88 / NF-κB signaling pathway and subsequent inhibition of inflammatory response, thus playing a role in the prevention and treatment of CKD.

Claims

1. A Chinese medicine composition for treating chronic kidney disease, characterized in that: It is prepared from the following raw materials in the following weight ratio: 4-6 parts of Cimicifuga, 24-36 parts of Rhubarb, 8-12 parts of Polygonum cuspidatum, 12-18 parts of Scutellaria baicalensis, 8-12 parts of Artemisia capillaris, 4-6 parts of Wax Gourd Peel, 4-6 parts of Leonurus japonicus, 4-6 parts of Leech, 4-6 parts of Pheretima, 4-6 parts of Carthamus tinctorius, and 4-6 parts of Millettia reticulata.

2. The Chinese medicine composition according to claim 1, characterized in that: It is prepared from the following raw materials in the following weight ratio: 5 parts of Cimicifuga, 30 parts of Rhubarb, 10 parts of Polygonum cuspidatum, 15 parts of Scutellaria baicalensis, 10 parts of Artemisia capillaris, 5 parts of Wax Gourd Peel, 5 parts of Leonurus japonicus, 5 parts of Leech, 5 parts of Pheretima, 5 parts of Carthamus tinctorius, and 5 parts of Millettia reticulata.

3. The Chinese medicine composition according to claim 1 or 2, characterized in that: The invention is prepared from the raw drug powder, water or organic solvent extract of the raw drug as active ingredients, and pharmaceutically acceptable auxiliary materials are added to prepare a commonly used intestinal administration preparation in medicine.

4. The Chinese medicine composition according to claim 3, characterized in that: The intestinal drug delivery preparations include enemas, suppositories, and intestinal foams.

5. A method for preparing the Chinese medicine composition according to any one of claims 1 to 4, characterized in that: It includes the following steps: a. Weigh the raw materials of each weight ratio and crush them; b. Add water and boil, concentrate the decoction, and add pharmaceutically acceptable excipients to prepare a commonly used enteral administration preparation in medicine.

6. Use of the Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a drug for enteral administration for treating chronic kidney disease.

7. The use according to claim 6, characterized in that: The drug is a drug that reduces creatinine, proteinuria, endotoxin, inflammatory factors, inhibits renal fibrosis, and delays the decline of renal function in chronic kidney disease by intervening in the gut-kidney axis and TLR4 / MyD88 / NF-κB pathway.

8. The use according to claim 6, characterized in that: The medicine is used to treat chronic kidney disease with water-gas syndrome or blood stasis syndrome.

9. Use of the Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a drug for intestinal administration having the effects of expelling turbidity and removing dampness, promoting blood circulation and promoting diuresis.

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