Abdomen skin pale-yellow soup and application thereof in treatment of knee osteoarthritis

Abstract

The invention provides an abdominal skin pinghuang decoction and application thereof in treatment of knee osteoarthritis, and belongs to the technical field of traditional Chinese medicines. The invention provides a traditional Chinese medicine composition for treating knee osteoarthritis, which comprises pericarpium arecae, glabrous sarcandra herb, semen brassicae, radix clematidis, poria peel, rhizoma atractylodis, cortex phellodendri and radix cyathulae, and can play a comprehensive treatment effect from multiple ways of resisting inflammation, easing pain, regulating cartilage metabolism, resisting oxidation, inhibiting cartilage cell apoptosis and the like through the synergistic effect of various active ingredients. The cartilage protection or repair is promoted while the symptoms are relieved, so that the defects of the existing western medicine treatment are overcome. According to the traditional Chinese medicine composition disclosed by the invention, structural damage, dysfunction and inflammation microenvironments are improved at the same time, the limitation of single-path treatment of celecoxib is broken through, and the curative effect of the traditional Chinese medicine composition exceeds that of celecoxib at a high dosage.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine, and particularly relates to a Fupi Canghuang Decoction and application thereof in the treatment of knee osteoarthritis. BACKGROUND

[0002] Osteoarthritis (OA) is a degenerative joint disease with high incidence and serious impact on the quality of life of patients, and its characteristics are progressive destruction of articular cartilage, subchondral bone sclerosis, osteophyte formation and synovial inflammation.

[0003] In the preclinical study of OA, the rat model induced by knee joint injection of monoiodoacetate (MIA) is widely recognized and applied. MIA specifically inhibits the activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in chondrocytes, interferes with the glycolysis process, causes energy metabolism disorder and death of chondrocytes, and then causes dose-dependent degradation of articular cartilage matrix, synovial inflammation and pain behavior. This model can better simulate the key pathophysiological characteristics of human OA, including cartilage damage, inflammatory response and pain, and is a reliable tool for evaluating the efficacy of potential OA treatment drugs.

[0004] At present, non-steroidal anti-inflammatory drugs (NSAIDs), such as selective COX-2 inhibitor celecoxib, are commonly used drugs for the treatment of OA pain and inflammation in clinical practice. However, the western medicine represented by celecoxib has significant limitations, including long-term use safety issues, potential cardiovascular risks (such as increasing the risk of myocardial infarction and stroke) and gastrointestinal adverse reactions (such as ulcer and bleeding), which limit its long-term and large-dose application in some patients. In contrast, traditional Chinese medicine (TCM) formulae have unique potential in treating complex diseases such as OA due to their multi-component, multi-target and overall regulation characteristics, so the use of TCM strategies to treat OA has become a major medical problem that needs to be solved urgently. SUMMARY

[0005] The application provides a Fupi Canghuang Decoction (FPCHT) and application thereof in the treatment of knee osteoarthritis. The FPCHT can exert comprehensive therapeutic effects through the synergistic effect of multiple active ingredients in multiple ways, relieve the symptoms of knee osteoarthritis, and promote cartilage protection or repair.

[0006] The application provides a traditional Chinese medicine composition for treating knee osteoarthritis, which comprises the following components in parts by mass: Fupi 15 parts, Zhongjie Feng 15 parts, Baijiezi 10 parts, Weilingxian 30 parts, Fulingpi 30 parts, Cangshu 10 parts, Huangbai 10 parts and Chuan Niuxi 15 parts.

[0007] The application also provides application of the traditional Chinese medicine composition in the preparation of a medicine for improving joint swelling.

[0008] The application further provides application of the Chinese medicine composition in preparation of a medicine for improving bipedal balance.

[0009] The application further provides application of the Chinese medicine composition in preparation of a medicine for improving the inflammatory microenvironment of arthritis.

[0010] In a preferred mode of the application, the improvement of the inflammatory microenvironment comprises improvement of the inflammatory microenvironment induced by IL-1β and / or IL-6.

[0011] The application further provides application of the Chinese medicine composition in preparation of a medicine for treating knee osteoarthritis.

[0012] In a preferred mode of the application, the dose of the Chinese medicine composition in the medicine is not less than 1.0 x HCED. The HCED is a human clinical equivalent dose based on body surface area.

[0013] The application further provides a medicine for treating knee osteoarthritis, comprising the Chinese medicine composition and a pharmaceutically acceptable excipient.

[0014] In a preferred mode of the application, the dosage form of the medicine comprises an oral dosage form.

[0015] In a preferred mode of the application, the concentration of crude drugs in the decoction of the Chinese medicine composition is not less than 1.35 g / mL.

[0016] Beneficial effects: The application provides a Chinese medicine composition for treating knee osteoarthritis. According to the theory of traditional Chinese medicine and clinical experience, Dabaipi, Zhongjiefeng, Baijiezi, Weilingshen, Fulingpi, Cangshu, Huangbai and Chuan Niuxi are combined to form a prescription. The comprehensive therapeutic effect can be achieved through the synergistic effect of multiple active ingredients in multiple pathways such as anti-inflammatory, analgesic, regulation of cartilage metabolism, antioxidant and inhibition of chondrocyte apoptosis. The symptoms can be relieved, and the cartilage protection or repair can be promoted, so as to make up for the deficiency of existing western medicine treatment.

[0017] The present application evaluates the pharmacodynamic effect of the Chinese medicine compound Fupeicanghuang Decoction on the knee osteoarthritis model induced by intra-articular injection of sodium monoiodoacetate (MIA) in rats by testing joint swelling degree, double foot balance and inflammatory factor detection. The joint swelling degree results show that the non-steroidal anti-inflammatory drug celecoxib can significantly reduce the knee joint swelling degree during the treatment period, the high-dose Chinese medicine compound has the same effect as celecoxib, and the Chinese medicine compound shows a dose-dependent inhibitory effect on the knee joint swelling induced by MIA. The double foot balance results show that the non-steroidal anti-inflammatory drug celecoxib can significantly improve the double foot balance during the treatment period, the high-dose Chinese medicine compound is better than celecoxib, and the Chinese medicine compound shows a dose-dependent improvement effect on the double foot balance induced by MIA. The inflammatory factor detection results show that the non-steroidal anti-inflammatory drug celecoxib can significantly inhibit the IL-1β and IL-6 levels in synovial fluid, the high-dose Chinese medicine compound has a significant inhibitory effect better than celecoxib, and the Chinese medicine compound shows a dose-dependent inhibitory effect on the inflammatory factor levels in synovial fluid or serum induced by MIA.

[0018] In summary, FPCHT improves structural damage (swelling), dysfunction (balance) and inflammatory microenvironment (IL-1β / IL-6) at the same time, breaking through the limitations of celecoxib single-path treatment. All pharmacodynamic indicators show a dose gradient response, providing accurate basis for clinical drug administration scheme. In the two key areas of functional recovery (balance improvement) and deep anti-inflammatory (cytokine inhibition), the high-dose group of FPCHT has better curative effect than the existing standard drug celecoxib. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 The results of joint swelling degree of rats are shown in the following figure; Figure 2 The results of foot balance of rats are shown in the following figure. DETAILED DESCRIPTION

[0020] The present application provides a Chinese medicine composition for treating knee osteoarthritis, which comprises the following components by mass: Dafupi 15 parts, Zhongjie wind 15 parts, Baijiezi 10 parts, Weilingxian 30 parts, Fulingpi 30 parts, Cangshu 10 parts, Huangbai 10 parts and Chuan Niuxi 15 parts.

[0021] The core pathogenesis of knee osteoarthritis synovitis is deficiency of both spleen and kidney, water and dampness stagnation, overflow of skin, and blockage of meridians by wind, cold and dampness. The treatment principle should be mainly to invigorate the spleen and kidney, promote qi and drain water, remove dampness and dredge collaterals. Therefore, the classical prescription of Guqing Decoction and Simiao Pills are used as the base prescription, and Fupeicanghuang Decoction is prepared by adding and subtracting. In this prescription, Fulingpi and Dafupi are the monarchs: the two are combined, one invigorating the spleen and draining water, and the other promoting qi and draining water, so that the water is removed from the small intestine. Wilingxian and Zhujiefeng are the ministers: Wilingxian is pungent and warm, and can remove wind and dampness, and relieve pain by removing wind and dampness and dredging collaterals. Zhujiefeng is bitter and cold, and can remove wind and blood, clear heat and resolve toxicity. The two combined can enhance the effect of removing wind and dampness, dredging collaterals and relieving pain, especially for treating swelling and pain of lower limbs. Cangshu, Baizhizi and Huangbai are the adjuvants: Cangshu can dry the spleen and dampness, and invigorating the spleen can prevent the regeneration of water and dampness from the source. Baizhizi is pungent and warm, and can remove dampness and phlegm, especially for eliminating dampness and phlegm in joint cavity. Huangbai is cold, which can prevent the accumulation of heat caused by large amount of pungent and warm herbs (such as Cangshu and Wilingxian), and can clear heat and remove dampness if the disease has the tendency of heat (such as local red and hot feeling), which is the meaning of “Ermiacao” (Cangshu+Huangbai) for treating dampness and heat in lower jiao. Chuan Niuxi is the minister: Chuan Niuxi can not only activate blood and dredge collaterals, but also guide other herbs to downward, so as to ensure that the medicinal effect is exerted on the lesions of lower limbs, knees and ankles. The whole prescription combines cold and heat, invigorates the spleen and kidney, dredges qi, and highlights the effects of removing dampness and phlegm, removing wind and dampness, dredging collaterals and relieving pain, which is a clinical special prescription for treating knee osteoarthritis.

[0022] The traditional Chinese medicine composition is a decoction, which can be obtained by soaking and decocting, and is called Fupeicanghuang Decoction (FPCHT). In the present application, the soaking is to add the above-mentioned medicinal materials into clean drinking water, and the water level is 2-3 cm higher than the medicinal materials, and the soaking time is 30-60 min.

[0023] The decocting is to first boil with strong fire and then change to moderate fire, so that the effective components are fully extracted, and finally concentrated to about 250 mL. In the present application, the Fupeicanghuang Decoction for patients is combined with Dafupi 15 g, Zhujiefeng 15 g, Baizhizi 10 g, Wilingxian 30 g, Fulingpi 30 g, Cangshu 10 g, Huangbai 10 g and Chuan Niuxi 15 g. In the rat experiment, the medicinal materials are 5 times of the patient's medicinal material, that is, the Fupeicanghuang Decoction for rats is combined with Dafupi 75 g, Zhujiefeng 75 g, Baizhizi 50 g, Wilingxian 150 g, Fulingpi 150 g, Cangshu 50 g, Huangbai 50 g and Chuan Niuxi 75 g, and the concentration of the prepared crude drug is 2.7 g / mL. 2.7 g / mL is the high-dose group of animals, the medium-dose group is 1.35 g / mL, and the low-dose group is 0.675 g / mL.

[0024] The present application also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a medicine for improving joint swelling.

[0025] In the embodiment of the present application, the MIA modeling results show that, after modeling, the right knee joint swelling of the MIA modeling group is significantly increased compared with the healthy group; compared with the MIA model control group, the knee joint swelling of the celecoxib positive drug group and the medium and high dose compound groups is significantly reduced after administration. The traditional Chinese medicine compound shows a dose-dependent inhibitory effect on the knee joint swelling induced by MIA.

[0026] The present application also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for improving bipedal balance.

[0027] In the embodiment of the present application, the MIA modeling results show that, after modeling, the bipedal balance (%) of the MIA modeling group is significantly increased compared with the healthy group; compared with the MIA model control group, the bipedal balance (%) of the celecoxib positive drug group and the low, medium and high dose compound groups is significantly reduced after administration. The traditional Chinese medicine compound shows a dose-dependent improvement effect on the bipedal balance induced by MIA.

[0028] The present application also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for improving the inflammatory microenvironment of arthritis.

[0029] The present application improves the inflammatory microenvironment, including improving the inflammatory microenvironment induced by IL-1β and / or IL-6. In the embodiment of the present application, the MIA modeling results show that, compared with the healthy group, the serum IL-1β, synovial fluid IL-1β and IL-6 of the MIA model control group are significantly increased; compared with the MIA model control group, the synovial fluid IL-1β and IL-6 of the celecoxib positive drug group and the low, medium and high dose compound groups are significantly reduced after administration; the serum IL-1β of the high dose compound group is significantly reduced. In addition, the traditional Chinese medicine compound shows a dose-dependent inhibitory effect on the synovial fluid IL-1β and IL-6 levels induced by MIA.

[0030] The present application also provides the use of the above-mentioned traditional Chinese medicine composition in the preparation of a drug for treating knee osteoarthritis.

[0031] In the embodiment of the present application, the MIA modeling results show that FPCHT simultaneously improves structural damage (swelling), dysfunction (balance) and inflammatory microenvironment (IL-1β / IL-6), breaking through the limitations of single-path treatment of celecoxib. And all the pharmacodynamic indexes show a dose gradient response, providing accurate basis for clinical administration scheme. In the two key areas of functional recovery (balance improvement) and deep anti-inflammatory (cytokine inhibition), the high dose group of FPCHT has better curative effect than the existing standard drug celecoxib. The dose of the traditional Chinese medicine composition in the drug is not less than 1.0×HCED; the HCED is the human clinical equivalent dose based on the body surface area.

[0032] The application further provides a medicine for treating knee osteoarthritis, comprising the traditional Chinese medicine composition and pharmaceutically acceptable adjuvants.

[0033] The dosage form of the medicine comprises oral dosage forms, and the concentration of crude drugs in the decoction of the traditional Chinese medicine composition is not less than 1.35 g / mL.

[0034] In order to further illustrate the application, the use of the Fupeicanghuang Decoction and in the treatment of knee osteoarthritis is described in detail below in combination with examples, but they should not be understood as limiting the protection scope of the application.

[0035] The reagents used in the examples of the application are all conventional reagents in the art, wherein: Modeling agent: sodium iodoacetate (MIA), supplier: Aladdin; batch number: 12402311; an appropriate amount of MIA was weighed and dissolved in normal saline to prepare a solution with a concentration of 2 mg / 50 μL; Positive drug: celecoxib, supplier: MCE; item number: HY-14398; batch number: 682067; an appropriate amount of celecoxib was weighed and dissolved in 0.5% sodium hydroxymethyl cellulose to prepare a solution with a concentration of 2.0 mg / mL, which was prepared once a day during the administration period; Fupeicanghuang Decoction: 15 g of Dafupi, 15 g of Zhongjiefeng, 10 g of Baijiezi, 30 g of Weilingxian, 30 g of Fulingpi, 10 g of Cangshu, 10 g of Huangbai, and 15 g of Chuanniuxi; the above-mentioned medicinal materials (5 doses) were placed in a fire-resistant ceramic sand pot, clean drinking water was added, and the water was soaked for 30 minutes, the water level was 2-3 cm higher than the medicinal materials; after boiling with strong fire, the effective components were fully extracted with weak fire, and finally concentrated to about 250 mL. At this time, the crude drug concentration of the compound decoction was about 2.7 g / mL. The concentrated liquid was used for high-dose compound group gavage in rats, with a gavage volume of 1 mL / 100 g of body weight. The high-dose compound decoction was diluted with pure water 1:1 for medium-dose group gavage, with a gavage volume of 1 mL / 100 g of body weight. The medium-dose compound decoction was diluted with pure water 1:1 for low-dose group gavage, with a gavage volume of 1 mL / 100 g of body weight. The high-dose compound decoction was stored at 2-8℃ for 7 days, and was boiled once a week; the medium-dose and low-dose compound decoctions needed to be prepared on demand during the administration period.

[0036] Experimental animals: Wistar rats, male, 6-8 weeks old; supplier: Beijing Vantoll Life Experimental Animal Technology Co., Ltd.

[0037] Modeling scheme: 7 days before modeling (D-7), the rats were anesthetized by inhaling isoflurane (manufacturer: Orbiepharm, batch number: 20241003), and 50 μL of MIA solution was injected into the right knee joint cavity for single injection. The first day of administration was D0, and the rats were sacrificed 21 days after administration (D21).

[0038] Detection index: Joint swelling degree quantitative analysis: On D-7, D-1 before administration and D7 / D14 / D21 after administration, the diameters of bilateral knee joints were measured using a vernier caliper. The right knee swelling degree was calculated by the formula: Swelling degree (%) = [(right knee diameter-left knee diameter) / left knee diameter] x 100.

[0039] Weight-bearing balance (double foot balance) behavioral evaluation: On D-7, D-1, D7, D14, D21, the foot balance tester (manufacturer: Ugo Basile, model: 47885) was used to detect the weight-bearing force of the bilateral lower limbs. The data was standardized as the weight-bearing balance ratio: Weight-bearing balance ratio (%) = (left weight-bearing force / right weight-bearing force) x 100.

[0040] Inflammatory factor detection (MSD multi-factor platform): Serum collection: At the end of the experiment (D21), after isoflurane anesthesia of the rats, 300 μL of whole blood was collected into an anticoagulant-free centrifuge tube by orbital venous plexus or jugular vein puncture. After standing at room temperature for 30 minutes, the serum was separated by centrifugation (5000g, 10 min) at 4°C and stored at -80°C for later use.

[0041] Synovial fluid collection: After anesthesia of the rats on D21, the joint cavity was punctured into the right knee joint under the patella, and 100 μL of normal saline containing 20 μg / mL indomethacin (manufacturer: Sigma-Aldrich, product number: 17378-5G) was injected. After repeated perfusion, the perfusate was withdrawn and stored at -80°C.

[0042] Detection target: The levels of TNF-α, IL-1β and IL-6 in serum and synovial fluid were quantitatively analyzed by MSD electrochemiluminescence technology (ultra-sensitive multi-factor chemiluminescence analyzer, manufacturer: PHCbi, model: SQ120MM).

[0043] Statistical analysis: All data were expressed as mean ± standard error (mean ± SEM). Hypothesis testing was performed using GraphPad Prism 8.0 software. Repeated measures ANOVA was used for joint swelling degree and weight-bearing balance ratio, and Bonferroni method was used for multiple comparisons between groups; One-way ANOVA was used for inflammatory factor data, and Dunnett method was used for comparison between groups. The significance threshold was set at P<0.05. Figure 1 and Figure 2Data are presented as mean ± SEM. Repeated Measures ANOVA was used to analyze the overall difference, followed by Bonferroni post-hoc test for comparison with G2 control group. Statistical significance was defined as *p<0.05, **P<0.01, ***P<0.001.

[0044] Example 1 I. Experimental grouping and dosing regimen According to the results of D-1 foot balance test and body weight, MIA model animals were randomly divided into 5 groups (G2~G6), 8 in each group: (1) G1 group: healthy group (N=6); (2) G2 group: MIA model control group (N=8, once a day by gavage with the same volume of normal saline); (3) G3 group: MIA+Celecoxib (positive drug) group (N=8, 20mg / kg, once a day by gavage); (4) G4 group: MIA+low-dose compound group (N=8, 6.75g / kg, once a day by gavage); (5) G5 group: MIA+medium-dose compound group (N=8, 13.5g / kg, once a day by gavage); (6) G6 group: MIA+high-dose compound group (N=8, 27g / kg, once a day by gavage).

[0045] Gavage volume is 1mL / 100g (rat weight), for example: 200g rat dosing volume = 2mL; Dose design basis: Traditional Chinese Medicine Compound Dose is based on Human Clinical Equivalent Dose (HCED) converted by body surface area, and three dose gradients are set up: (a) Low-dose group: 0.5x HCED; (b) Medium-dose group: 1.0x HCED; (c) High-dose group: 2.0x HCED.

[0046] II. Experimental results 2.1 Joint swelling results Before modeling D-7, before administration D-1, after administration D7, D14 and D21, the left and right knee joint diameters of rats were detected, and the right knee joint swelling was calculated.

[0047] Joint swelling results are shown in Figure 1Tables 1 and 2. After modeling, the swelling of the right knee joint in the MIA model group was significantly increased compared with the healthy group; compared with the MIA model control group, the knee joint swelling was significantly reduced in the celecoxib positive drug group and the medium and high dose compound groups after administration. The traditional Chinese medicine compound showed a dose-dependent inhibitory effect on MIA-induced knee joint swelling.

[0048] Table 1. Joint swelling results (%) - Mean±SEM

[0049] Table 2. Joint swelling degree P-value

[0050] 2.2 Foot balance results The weight-bearing capacity of the rats' hind limbs was tested on D-7 before modeling, D-1 before drug administration, and D7, D14 and D21 after drug administration to evaluate the rats' behavior before and after drug administration; foot balance was represented by the ratio of weight-bearing capacity of the left foot to weight-bearing capacity of the right foot * 100%.

[0051] The results are as follows Figure 2 As shown in Tables 3 and 4, the foot balance data for each group of animals are presented. After modeling, the foot balance (%) in the MIA model group was significantly increased compared with the healthy group; compared with the MIA model control group, the foot balance (%) in the celecoxib positive drug group and the low, medium, and high dose compound groups was significantly decreased after administration. The traditional Chinese medicine compound showed a dose-dependent improvement effect on MIA-induced bipedal balance.

[0052] Table 3 Foot balance results (%) - Mean±SEM

[0053] Table 4. Foot balance P-value

[0054] 2.3 Factor detection results At the experimental endpoint, rat serum and joint lavage fluid were collected. All samples were tested for inflammatory factors TNF-α, IL-1β, and IL-6.

[0055] The results of factor detection are shown in Tables 5 and 6. Serum IL-1β, synovial fluid IL-1β, and IL-6 were significantly increased in the MIA model control group compared to the healthy group. Compared to the MIA model control group, synovial fluid IL-1β and IL-6 were significantly decreased in the celecoxib-positive drug group and the low, medium, and high-dose compound groups after administration; serum IL-1β was significantly decreased in the high-dose compound group. Furthermore, the traditional Chinese medicine compound showed a dose-dependent inhibitory effect on MIA-induced synovial fluid IL-1β and IL-6 levels.

[0056] Table 5 Factor detection results (pg / mL) - Mean ± SEM

[0057] Table 6 Factor detection P values

[0058] In summary, FPCHT improved structural damage (swelling), functional impairment (balance), and inflammatory microenvironment (IL-1β / IL-6) simultaneously, breaking the limitation of celecoxib monotherapy. In both functional recovery (balance improvement) and deep anti-inflammation (cytokine inhibition), the high-dose FPCHT group outperformed the existing standard drug celecoxib.

[0059] Although the above embodiments have made a detailed description of the present application, it is only a part of the embodiments of the present application, not all the embodiments, and other embodiments can be obtained according to the present embodiments without creativity, which belong to the protection scope of the present application.

Claims

1. A traditional Chinese medicine composition for treating knee osteoarthritis, characterized in that, The medicine comprises the following quality parts: Dabaipi 15 parts, Zhongjie Feng 15 parts, Baijiezi 10 parts, Weilingxian 30 parts, Fulingpi 30 parts, Cangshu 10 parts, Huangbai 10 parts and Chuan Niuxi 15 parts.

2. Use of the traditional Chinese medicine composition of claim 1 in the preparation of a medicine for improving joint swelling.

3. Use of the traditional Chinese medicine composition of claim 1 in the preparation of a medicine for improving bipedal balance.

4. Use of the traditional Chinese medicine composition of claim 1 in the preparation of a medicine for improving the inflammatory microenvironment of arthritis.

5. Use according to claim 4, characterized in that, The improvement of the inflammatory microenvironment includes improvement of the inflammatory microenvironment induced by IL-1β and / or IL-6.

6. Use of the traditional Chinese medicine composition of claim 1 in the preparation of a medicine for treating knee osteoarthritis.

7. Use according to claim 6, characterized in that, The dose of the traditional Chinese medicine composition in the medicine is not less than 1.0×HCED; The HCED is the human clinical equivalent dose based on the body surface area.

8. A medicament for treating knee osteoarthritis, characterized by, The medicine comprises the traditional Chinese medicine composition of claim 1 and pharmaceutically acceptable excipients.

9. The medicament according to claim 8, characterized in that, The dosage form of the medicine comprises oral dosage.

10. The medicament according to claim 8 or 9, characterized in that, The concentration of crude drugs in the decoction of the traditional Chinese medicine composition is not less than 1.35 g / mL.

Images