Composition for treating hyperuricemia nephropathy and application thereof

Abstract

The invention discloses a composition for treating hyperuricemia nephropathy and application thereof, and belongs to the field of biological medicines.The composition is prepared from, by weight, 28-32 parts of rhizoma smilacis glabrae, 28-32 parts of yam rhizome, 13-17 parts of radix clematidis, 8-12 parts of peach kernels, 8-12 parts of flos carthami, 8-12 parts of herba lycopi, 18-22 parts of rhizoma alismatis, 28-32 parts of raw coix seeds, 28-32 parts of semen plantaginis, 13-17 parts of rhizoma atractylodis and 13-17 parts of pseudobulbus cremastrae seu pleiones. The composition can obviously reduce the uric acid level of a mouse with high uric acid nephropathy and improve kidney injury; meanwhile, intestinal flora of mice with the hyperuricemia nephropathy can be remarkably remodeled, abundance of beneficial bacteria Ackerman's bacteria is up-regulated, abundance of opportunistic pathogenic bacteria such as escherichia coli-shigella, desulfurization vibrio and enterococcus is down-regulated, and important significance is achieved for research and drug development of the hyperuricemia nephropathy.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of biological medicine, and particularly relates to a composition for treating hyperuricemic nephropathy and application thereof. BACKGROUND

[0002] Hyperuricemic nephropathy (HN) is a metabolic kidney disease characterized by progressive renal function impairment caused by long-term hyperuricemia (HUA). With the prevalence of global high-purine diet, obesity and other metabolic problems, the prevalence of HUA and its related kidney diseases has increased dramatically, making it the fourth metabolic disease after hypertension, hyperlipidemia and hyperglycemia. According to statistics, the prevalence of HUA in different countries and regions of the world is between 8.4% and 71.6%, and the prevalence of the disease is becoming younger. According to the 2021 China Hyperuricemia and Gout Trend White Paper, the prevalence of hyperuricemia is 13.3%, and the number of patients has reached 190 million. Continuous high uric acid status can lead to sodium urate crystal deposition, renal tubular injury, interstitial fibrosis and uric acid stone formation, eventually progressing to chronic kidney disease, even end-stage renal disease, which seriously affects the prognosis of patients. At present, the clinic mainly relies on allopurinol, febuxostat and other uric acid-lowering drugs, but long-term use of these drugs can cause adverse reactions such as hypersensitivity, liver toxicity and increased risk of cardiovascular disease, so it is urgent to develop safer and more effective therapeutic drugs.

[0003] Intestinal flora plays a key role in regulating host metabolism and maintaining intestinal homeostasis, and is the core link of the intestinal-renal axis interaction. The intestinal flora of patients with hyperuricemia shows a decrease in species richness and diversity, and the change in flora composition is related to disease progression. Intestinal flora remodeling, including the regulation of Akkermansia, Escherichia-Shigella, Desulfovibrio and Enterococcus, is closely related to hyperuricemic nephropathy; intestinal flora is not only a key regulatory factor of HN pathology, but also a promising treatment breakthrough due to its metabolic plasticity. SUMMARY

[0004] Therefore, the application discloses a composition for treating hyperuricemic nephropathy and application thereof.

[0005] The application adopts the following technical scheme:

[0006] The composition for treating hyperuricemic nephropathy comprises the following raw materials in parts by weight: 28-32 parts of Smilax glabra, 28-32 parts of Alisma orientale, 13-17 parts of Cynanchum wilfordii, 8-12 parts of Semen Persicae, 8-12 parts of Carthamus tinctorius, 8-12 parts of Lycopi Herba, 18-22 parts of Alisma orientale, 28-32 parts of Coix lachryma-jobi, 28-32 parts of Plantago asiatica, 13-17 parts of Atractylodes lancea, and 13-17 parts of Omphalotrichum.

[0007] Further, the composition comprises the following raw materials by weight: 30 parts of smilax glabra, 30 parts of yam, 15 parts of radix dipsaci, 10 parts of persicae semen, 10 parts of safflower, 10 parts of lycianthes, 20 parts of alisma, 30 parts of coicis semen, 15 parts of atractylodes, and 15 parts of ophiocordyceps.

[0008] Further, the preparation steps of the composition comprise:

[0009] S1 weighing the raw materials by weight;

[0010] S2 adding 10 times the weight of pure water to the raw materials and soaking for 1 hour;

[0011] S3 heating the soaking liquid to boiling with a small fire, and maintaining boiling for 45 minutes to obtain an extracted medicinal liquid;

[0012] S4 filtering the extracted medicinal liquid with double-layer gauze to obtain a filtrate and a residue;

[0013] S5 concentrating the filtrate to obtain a water decoction;

[0014] S6 freezing and storing the water decoction for standby use.

[0015] Further, in the residue of step S4, an appropriate amount of pure water is continuously added, boiled and maintained boiling for 30 minutes, and then filtered to obtain a second filtrate, and the second filtrate and the filtrate of step S4 are combined to obtain a mixed filtrate, and the mixed filtrate is concentrated to obtain the water decoction of step S5.

[0016] Further, the concentration of the water decoction of step S5 is 1.5 g / mL, and the concentration is calculated based on the crude drug amount.

[0017] Further, the freezing storage temperature of step S6 is-20°C.

[0018] Further, the use amount of the water decoction is 4-16 g / kg.

[0019] The application of a composition for treating hyperuricemic nephropathy, wherein the composition is used for regulating intestinal flora.

[0020] Further, the regulation of intestinal flora comprises remodeling intestinal flora of hyperuricemic nephropathy mice.

[0021] Further, the remodeling of intestinal flora of hyperuricemic nephropathy mice comprises: up-regulating the abundance of beneficial bacteria Akkermansia, and down-regulating the abundance of harmful bacteria Escherichia-Shigella, Desulfovibrio, and Enterococcus.

[0022] The beneficial effects of the present application are:

[0023] The high uric acid nephropathy composition disclosed by the application can significantly alleviate the weight loss of HN mice, down-regulate the kidney index and the spleen index, reduce the uric acid, creatinine and urea levels of HN mice, and improve the kidney injury; the composition can also significantly reshape the intestinal flora of HN mice, up-regulate the abundance of beneficial bacteria Akkermansia, and down-regulate the abundance of harmful bacteria such as Escherichia-Shigella, Desulfovibrio or Enterococcus, which has important significance for the research and drug development of high uric acid nephropathy. BRIEF DESCRIPTION OF DRAWINGS

[0024] In order to more clearly illustrate the technical solutions of the embodiments of the application, the drawings needed in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description are only some embodiments of the application, and other drawings can be obtained by those skilled in the art without creative labor.

[0025] Figure 1 The schematic diagram of the high uric acid nephropathy composition of the application reducing the plasma uric acid level of high uric acid nephropathy mice and improving the kidney injury;

[0026] Figure 2 The schematic diagram of the high uric acid nephropathy composition of the application reshaping the intestinal flora structure of high uric acid nephropathy mice;

[0027] Figure 3 The schematic diagram of the high uric acid nephropathy composition of the application significantly reversing the abundance of Akkermansia, Escherichia-Shigella, Desulfovibrio and Enterococcus. DETAILED DESCRIPTION

[0028] In order to better understand the technical solutions of the application, the embodiments of the application will be described in detail below with reference to the drawings.

[0029] It should be clear that the described embodiments are only some embodiments of the application, not all embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the application.

[0030] Embodiment 1

[0031] A high uric acid nephropathy treatment composition (TFF) comprises the following raw materials by weight: 28-32 parts of Smilax glabra, 28-32 parts of Yam, 13-17 parts of Clematis finetiana, 8-12 parts of Persicae Semen, 8-12 parts of Carthamus tinctorius, 8-12 parts of Lycopi Herba, 18-22 parts of Alisma orientale, 28-32 parts of Coicis Semen, 28-32 parts of Plantaginis Herba, 13-17 parts of Atractylodes lancea, and 13-17 parts of Omphalia lapatula.

[0032] Further, the composition comprises the following raw materials by weight: 30 parts of smilax glabra, 30 parts of yam, 15 parts of radix glehniae, 10 parts of semen pruni, 10 parts of safflower, 10 parts of lycianthes, 20 parts of alisma, 30 parts of coix, 15 parts of atractylodes, and 15 parts of dictyophora.

[0033] Further, the preparation steps of the composition comprise:

[0034] S1 weighing the raw materials by weight;

[0035] S2 adding 10 times the weight of pure water to the raw materials and soaking for 1 hour;

[0036] S3 heating the soaking liquid to boiling with gentle heat for 45 minutes to obtain an extracted medicinal liquid;

[0037] S4 filtering the extracted medicinal liquid with double-layer gauze to obtain a filtrate and a residue;

[0038] S5 concentrating the filtrate to obtain a water decoction;

[0039] S6 freezing and storing the water decoction for later use.

[0040] Further, the residue in step S4 is continuously added with an appropriate amount of pure water, which can be 8 times the weight of the raw materials, boiled and maintained boiling for 30 minutes, and then filtered to obtain a second filtrate. The second filtrate and the filtrate in step S4 are combined to obtain a mixed filtrate, and the mixed filtrate is concentrated to obtain the water decoction in step S5.

[0041] Further, the concentration of the water decoction in step S5 is 1.5 g / mL, which is calculated based on the weight of crude drugs.

[0042] Further, the freezing storage temperature in step S6 is -20°C.

[0043] Further, the use amount of the water decoction is 4-16 g / kg.

[0044] An application of a composition for treating hyperuricemic kidney disease, the above-mentioned composition is used for the regulation of intestinal flora.

[0045] Further, the regulation of intestinal flora includes remodeling the intestinal flora of hyperuricemic kidney disease mice.

[0046] Further, the remodeling of the intestinal flora of hyperuricemic kidney disease mice includes up-regulating the abundance of beneficial bacteria Akkermansia, and down-regulating the abundance of harmful bacteria such as Escherichia-Shigella, Desulfovibrio, and Enterococcus.

[0047] Example 2

[0048] High uric acid nephropathy composition (TFF) composition and preparation: Smilax 30g, Rhizoma 30g, Wuling San 15g, Peach kernel 10g, Red flower 10g, Herba 10g, Alisma 20g, Semen 30g, Atractylodes 15g, Yam 15g, a total of 11 kinds of medicine composition. All medicinal materials are added with 10 times the volume of double distilled water and soaked for 1 hour, and then boiled with gentle fire for 45 minutes. The filtrate is collected by double-layer gauze filtration, and then 8 times the volume of double distilled water is added and boiled for 30 minutes. The filtrate is collected by filtration, and the filtrate of two times is combined and concentrated by rotary evaporation. The concentrated water decoction is obtained, and the water decoction concentration is 1.5g / mL (calculated by crude drug), which is stored at -20℃ for use.

[0049] Example 3

[0050] High uric acid nephropathy composition (TFF) composition and preparation: Smilax 30g, Rhizoma 30g, Wuling San 15g, Peach kernel 10g, Red flower 10g, Herba 10g, Alisma 20g, Semen 30g, Atractylodes 15g, Yam 15g, a total of 11 kinds of medicine composition. All medicinal materials are added with 10 times the volume of double distilled water and soaked for 1 hour, and then boiled with gentle fire for 45 minutes. The filtrate is collected by double-layer gauze filtration, and then 8 times the volume of double distilled water is added and boiled for 30 minutes. The filtrate is collected by filtration, and the filtrate of two times is combined and concentrated by rotary evaporation. The concentrated water decoction is obtained, and the water decoction concentration is 1.5g / mL (calculated by crude drug), which is stored at -20℃ for use.

[0051] Example 4

[0052] High uric acid nephropathy composition (TFF) composition and preparation: Smilax 30g, Rhizoma 30g, Wuling San 15g, Peach kernel 10g, Red flower 10g, Herba 10g, Alisma 20g, Semen 30g, Atractylodes 15g, Yam 15g, a total of 11 kinds of medicine composition. All medicinal materials are added with 10 times the volume of double distilled water and soaked for 1 hour, and then boiled with gentle fire for 45 minutes. The filtrate is collected by double-layer gauze filtration, and then 8 times the volume of double distilled water is added and boiled for 30 minutes. The filtrate is collected by filtration, and the filtrate of two times is combined and concentrated by rotary evaporation. The concentrated water decoction is obtained, and the water decoction concentration is 1.5g / mL (calculated by crude drug), which is stored at -20℃ for use.

[0053] Example 5

[0054] Animal experiment: High uric acid nephropathy composition (TFF) improves the efficacy of high uric acid nephropathy drug research experiment, 8-week-old male C57BL / 6J mice, purchased from Liaoning Changsheng Biotechnology Co., Ltd., and fed in IVC mouse system. All mice were fed under standard conditions (light / dark cycle 12-12 hours, humidity 55±5%, temperature 23±2℃), free diet and drinking water, and the detailed experimental process is shown in Figure 2A. After one week of adaptive feeding, mice were randomly divided into 6 groups (n=6-8): Ctrl group, HN group (hyperuricemia nephropathy group), HN+All (allopurinol) group and HN+TFF-L / M / H group; mice in each group were given normal animal feed and sterilized distilled water. Mice in HN group, allopurinol group (HN+All) and TFF-low, medium and high dose groups (HN+TFF-L / M / H) were first given 100 mg / kg potassium oxonate (PO) and 37.5 mg / kg adenine hydrochloride suspension by gavage every day, and the solvent was 0.5% sodium carboxymethylcellulose solution. After an interval of 1 hour, the HN+All group was given 20 mg / kg allopurinol by gavage, and the HN+TFF-L / M / H were given 4 / 8 / 16 g / kg TFF decoction by gavage respectively. The Ctrl group was given an equal amount of solvent by gavage every day. Mice were killed 2 hours after the last administration ( Figure 1 A). The TFF decoction used in this example was the decoction obtained by the method of Example 2.

[0055] During the animal experiments, typical physiological parameters of the mice were monitored, and changes in their weight and water intake were regularly recorded. At the conclusion of the experiment, the mice were euthanized, and plasma, major tissue, and fecal samples were collected. Partial kidney tissue was fixed in 4% paraformaldehyde solution, and the remaining samples were stored at -80°C for further analysis. Animal experiments were conducted in accordance with the National Laboratory Animal Use Act and the regulations of the Hubei University of Chinese Medicine Ethics Committee.

[0056] Experimental results analysis:

[0057] 1. The hyperuric acid nephropathy composition reduces the uric acid level in hyperuric acid nephropathy mice and improves kidney damage.

[0058] like Figure 1 As shown in A, we first evaluated the improvement effect of 4g / kg, 8g / kg and 16g / kg of TFF decoction on hyperuricemia nephropathy induced by potassium oxonate combined with adenine. Figure 1 As shown in Figures B and 1C, All intervention significantly alleviated the weight loss of hyperuricemia nephropathy mice (P<0.05, vs. HN group). The effects of low- and medium-dose TFF intervention were similar to those of the positive drug All (P<0.05, vs. HN group). The kidney index and spleen index of hyperuricemia nephropathy mice increased significantly, while the high- and medium-dose hyperuricemia nephropathy compositions significantly decreased the kidney index, and the high- and medium-dose hyperuricemia nephropathy compositions significantly decreased the spleen index (P<0.05 or 0.01, vs. HN group) ( Figure 1D-1E). Plasma uric acid, creatinine and urea levels are important indicators of kidney function. Compared with the hyperuricemic nephropathy group, low and medium doses of hyperuricemic nephropathy composition treatment can significantly reduce the plasma uric acid, creatinine and urea levels of hyperuricemic nephropathy mice (P < 0.05 or 0.01, vs. HN group) Figure 1 F-1H). In addition, the color of the kidneys of hyperuricemic nephropathy mice is slightly pale, yellow, and the surface is wrinkled and not smooth. After the intervention of hyperuricemic nephropathy composition, the abnormal morphology of the kidney is improved Figure 1 I). H&E staining results show that the renal tubular epithelial cells of hyperuricemic nephropathy model mice appear vacuolar degeneration, the renal tubules are significantly dilated, the glomeruli are atrophic, and inflammatory cell infiltration appears in the renal interstitium. After the intervention of hyperuricemic nephropathy composition, the pathological changes of mice are improved, the renal tubular dilatation is significantly relieved, and the inflammatory cell infiltration area is reduced, which is similar to the HN+All group Figure 1 J). The above results suggest that the hyperuricemic nephropathy composition can reduce uric acid levels, restore water and salt metabolism function, and improve kidney damage.

[0059] 2. Hyperuricemic nephropathy composition reshapes the intestinal flora structure of hyperuricemic nephropathy mice, which is significantly reversed at the genus level.

[0060] 16S rDNA analysis showed that hyperuricemic nephropathy composition intervention significantly reversed the intestinal flora alpha diversity of hyperuricemic nephropathy mice, Chao1 index, Shannon index Figure 2 A-2B) (P < 0.05 vs. HN group). PCA analysis showed that the Ctrl group, HN group, and HN+TFF group formed a clear cluster separation Figure 2 C). In terms of flora structure, hyperuricemic nephropathy composition can reverse the up-regulation of Desulfobacterota and Proteobacteria caused by hyperuricemic nephropathy, and the significant increase of Firmicutes / Bacteroidetes ratio (F / B) Figure 2 D). At the family level, hyperuricemic nephropathy composition up-regulated Akkermansiaceae and Bacteroidaceae, while down-regulated Clostridia_UCG-014 and Ruminococcaceae Figure 2 E).

[0061] At the genus level, TFF significantly reversed the down-regulation of beneficial bacteria Akkermansia caused by HN Figure 3 A), and the up-regulation of harmful bacteria Escherichia-Shigella, Desulfovibrio, and Enterococcus Figure 3B). Specifically, the relative abundance of Akkermansia in the HN group mice was 0.3 times that of the Ctrl group, while the relative abundance of Akkermansia after TFF administration was restored to 3 times that of the HN group. The relative abundance of Escherichia-Shigella was 72 times that of the Ctrl group, and after TFF intervention, the relative abundance of the genus Escherichia-Shigella in the TFF administration group was 0.02 times that of the HN group. Similarly, the Desulfovibrio in the HN group mice was 11.97 times that of the Ctrl group, and the abundance of Desulfovibrio in the TFF administration group decreased to 0.1 times that of the HN group. Similarly, TFF administration can also significantly down-regulate the abundance of Enterococcus.

[0062] The traditional Chinese medicine composition disclosed in the present application can significantly reshape the intestinal flora of HN mice, up-regulate the beneficial bacteria Akkermansia, and down-regulate the abundance of harmful bacteria such as Escherichia-Shigella, Desulfovibrio, and Enterococcus.

[0063] The above has described the embodiments of the present application in detail, and the principles and implementation modes of the present application have been described by applying specific examples; the above description of the embodiments is only used to help understand the method of the present application and its core idea; at the same time, for those skilled in the art, according to the idea of the present application, the specific implementation mode and application range will be changed, and the above description should not be understood as a limitation of the present application.

Claims

1. A composition for treating hyperuricemia nephropathy, characterized in that: The composition comprises the following raw materials in parts by weight: 28-32 parts of Smilax glabra, 28-32 parts of Radix Dioscoreae, 13-17 parts of Radix Clematidis, 8-12 parts of peach kernel, 8-12 parts of safflower, 8-12 parts of Herba Eupatorii, 18-22 parts of Rhizoma Alismatis, 28-32 parts of raw Coix seeds, 28-32 parts of Plantago seeds, 13-17 parts of Atractylodes and 13-17 parts of Trillium.

2. The composition according to claim 1, characterized in that The composition comprises the following raw materials in parts by weight: 30 parts of Smilax glabra, 30 parts of Radix Dichroae, 15 parts of Radix Clematidis, 10 parts of peach kernel, 10 parts of safflower, 10 parts of Herba Eupatorii, 20 parts of Rhizoma Alismatis, 30 parts of raw Coix seeds, 30 parts of Plantago seeds, 15 parts of Atractylodes and 15 parts of Trillium.

3. The composition according to claim 2, characterized in that The preparation steps of the composition include: S1 weighs the raw materials in parts by weight; S2. Add 10 times the weight of pure water to the raw material and soak for 1 hour; S3: The soaking liquid is heated to boiling over a slow fire and the boiling is continued for 45 minutes to obtain an extracting liquid; S4: filtering the extracted medicinal solution through a double-layer gauze to obtain a filtrate and a filter residue; S5: concentrating the filtrate to obtain a decoction; S6: freezing and storing the decoction for future use.

4. The composition according to claim 3, characterized in that Continue to add an appropriate amount of pure water to the filter residue in step S4, boil and maintain boiling for 30 minutes, then filter to obtain a secondary filtrate, combine the secondary filtrate and the filtrate in step S4 to obtain a mixed filtrate, and concentrate the mixed filtrate to obtain the water decoction in step S5.

5. The composition according to claim 4, characterized in that The concentration of the water decoction in step S5 is 1.5 g / mL, which is calculated based on the amount of the crude drug.

6. The composition according to claim 5, characterized in that The temperature of the frozen storage in step S6 is -20°C.

7. The composition according to claim 6, characterized in that The usage amount of the water decoction is 4-16g / kg.

8. An application of a composition for treating hyperuricemia nephropathy, characterized in that: The composition according to any one of claims 1 to 7 is used for regulating intestinal flora.

9. The application according to claim 8, characterized in that: The regulation of the intestinal flora includes reshaping the intestinal flora of hyperuricemia nephropathy mice.

10. The use according to claim 9, characterized in that: The method of reshaping the intestinal flora of hyperuricemia nephropathy mice includes: upregulating the abundance of Akkermansia and downregulating the abundance of Escherichia coli-Shigella, Desulfovibrio or Enterococcus.

Images