Effect and Mechanism of Qianggu Tongbi Capsule on Osteoarthritis Factors in Rats / 华中科技大学学报(医学版)

Objective To investigate the effect and mechanism of Qianggu Tongbi capsule on osteoarthritis factors in rats.Methods Osteoarthritis(OA)rat model was established by anterior cruciate ligament transection method.Rats were ran-domly divided into model group,Qianggu Tongbi capsule group and glucosamine group.HE staining was used to identify the model.Toluidine blue staining was used to observe the changes of cartilage.Real-time quantitative polymerase chain reaction(qRT-PCR)was applied to detect the expression of Toll-like receptors(TLR4)and nuclear factor kappa-B(NF-κB)in each group.The levels of IL-1β,TNF-α,and IL-6 were measured by the enzyme linked immunosorbent assay(ELISA)kit.The levels of TLR4 and p-NF-κB were detected by immunohistochemistry.Results HE staining showed that,compared with the control group,a large number of inflammatory cell infiltration and synovial tissue thickening were observed in the model group,while the inflammatory cell infiltration and synovial tissue thickening were not obvious in the Qianggu Tongbi capsule and glu-cosamine groups.Toluidine blue staining results showed that chondroitin sulfate in the cartilage tissue of the model group was significantly reduced compared with the control group,while levels of chondroitin sulfate in the Qianggu Tongbi capsule group and glucosamine group were significantly increased.Compared with control group,the expression and immunohistochemical re-sults of TLR4 and NF-κB,and the levels of IL-1β,TNF-α and IL-6 in model group were significantly increased(all P＜0.05).Compared with model group,the expression and immunohistochemical results of TLR4 and NF-KB,and the levels of IL-1β,TNF-α and IL-6 in Qianggu Tongbi capsule group and glucosamine group were significantly decreased(all P＜0.05).Conclusion Qianggu Tongbi capsule can inhibit expression of TLR4 and NF-κB by reducing the production of inflammatory factors to a-chieve the effect of treatmenting osteoarthritis.

Protective Effect of Cangxi Tongbi Capsule on Articular Cartilage of KOA Rats Based onp38 MAPK Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the therapeutic effect and mechanism of Cangxi Tongbi capsule on knee osteoarthritis (KOA) model rats. Method:Sixty 4-week-old SPF healthy male SD rats were randomly divided into blank group, model group, dimethyl sulfoxide (DMSO) group, Cangxi Tongbi capsule group, SB203580 group and Cangxi Tongbi capsule combined with SB203580 group. In addition to the normal group, the modified Hulth method was used to establish the koa model. After the model was established successfully, the Cangxi Tongbi capsule group was given 0.25 g·kg-1 Cangxi Tongbi capsule solution by gavage every day, the SB203580 group was given 0.015 g·kg-1 SB203580 solution by gavage, the Cangxi Tongbi capsule combined with SB203580 group was given a mixed solution containing 0.015 g·kg-1 SB203580 and 0.25 g·kg-1 Cangxi Tongbi capsule by gavage, the DMSO group was given 1% DMSO solution by gavage, the model group and blank group were given normal saline by gavage The stomach was killed and the material was taken after 4 weeks of drug intervention. The expression levels of interleukin-1β （IL-1β） and tumor necrosis factor-α（TNF-α）in the supernatant of peripheral blood were detected by ELISA. p38, p-p38, matrix metalloproteinase-13(MMP-13), Collagen Ⅱ mRNA and protein in p38 MAPK signal pathway were detected by Real-time fluorescence quantification PCR （Real-time PCR） and Western blot, and the localization expression of p-p38 was detected by immunohistochemistry. Result:Compared with normal group，the expression levels of p38，p-p38，MMP-13 in articular cartilage of the model group were up-regulated (P<0.01), the expression levels of CollagenⅡ was down-regulated (P<0.01). The contents of IL-1β and TNF-α in serum were significantly increased (P<0.01). Compared with model group, the expression levels of p38，p-p38，MMP-13 in articular cartilage of the Cangxi Tongbi capsule group, SB203580 group and Cangxi Tongbi capsule combined with SB203580 group were down-regulated (P<0.01), the expression levels of CollagenⅡ was up-regulated (P<0.01). The contents of IL-1β and TNF-α in serum were significantly decreased (P<0.01). Conclusion:Cangxi Tongbi capsule can effectively protect the cartilage of KOA rats, and its mechanism may be related to the targeted blocking of p38 MAPK signal pathway.

The clinical study of Tongbi capsule combined with acupuncture in the treatment of knee osteoarthritis / 国际中医中药杂志

Objective To explore the clinical effect of Tongbi capsule combined with acupuncture for the patients with knee osteoarthritis and explore the mechanism of inflammatory response and hemorrheology. Methods According to the random table method, 88 KOA patients in our hospital from February 2016 to January 2017 were divided into the control group and the research group with 44 in each group. Patients in the control group were treated bycelecoxib capsule, while the patients in the research group were treated by Tongbi capsule combined with acupuncture. The treatment courses of two groups were two months. The clinical total effective rate of two groups of patients were compared after treatment. At the same time, the osteoarthritis index visual scale (WOMAC) and morning stiffness, joint tenderness index score were detected and compared after the treatment.The levels of serum tumor necrosis factor-α(TNF-α), interleukin (IL)-1β and IL-6 were detected by enzyme linked immunosorbent assay before and after the treatment. The hemorheology indexes were tested by automatic biochemical analyzer. In addition, the incidence of adverse reactions of two groups of patients was observed during treatment. Results The clinical total effective rate of the research group was 93.18％ (41/44), which was significantly higher than the control group72.73％ (32/44), and the difference was statistically significant (χ2=6.510, P=0.011). After treatment, the WOMAC (31.26 ± 6.12 vs. 36.17 ± 7.09, t=-3.477), joint tenderness index (1.75 ± 0.46 vs. 2.11 ± 0.54, t=-3.366) and morning stiffness score (1.62 ± 0.53 vs. 2.14 ± 0.51, t=-4.690) of the research group were significantly lower than those of the control group (P<0.05). The levels of TNF-α (7.36 ± 1.45 pg/ml vs. 8.02 ± 1.54 pg/ml, t=-2.070), IL-1β (27.82 ± 5.22 μg/ml vs. 33.05 ± 5.17 μg/ml, t=-4.722), IL-6 (2.81 ± 0.58 ng/ml vs. 3.34 ± 0.53 ng/ml, t=-4.475) of the research group were significantly lower than those of the control group (P<0.05). The whole blood high shear viscosity (12.09 ± 3.46 mPa?s vs. 14.22 ± 3.51 mPa?s, t=-2.867), whole blood low shear viscosity (2.53 ± 0.68 mPa?s vs. 3.32 ± 0.71 mPa?s, t=-5.330), whole blood viscosity (1.35 ± 0.29 mPa?s vs. 1.60 ± 0.41 mPa?s, t=-3.302), erythrocyte sedimentation rate (7.10 ± 2.06 mm/h vs. 8.02 ± 2.13 mm/h, t=-2.059), fibrinogen (2.71 ± 0.53 g/L vs. 3.42 ± 0.65 g/L, t=-5.615) and erythrocyte deformation index (0.57 ± 0.16 vs. 0.71 ± 0.19, t=-3.739) of the research group were significantly lower than those of the control group (P<0.05). The incidence of adverse reactions of control group was 11.37％ (5/44), while the incidence of adverse reactions of resarch group was 6.82％ (3/44), which the difference of the incidence of adverse reactions between two groups showed no statistical significance (χ2=0.550, P=0.458). Conclusions The Tongbi capsule combined with acupuncture can reduce the degree of inflammation, and also improve hemorheology indexes of KOA patients.

[Effects of Tongbi capsule on joint lesions in rabbits with rheumatoid arthritis].

The purpose of this experiment is to observe the effects of Tongbi capsule on joint lesions in rabbit with rheumatoid arthritis induced by ovalbumin and explore the mechanism in order to provide reference for clinical application of Tongbi capsule. Rheumatoid arthritis in rabbits was induced by subcutaneous injection of emulsions of ovalbumin and Freund's complete adjuvant and intra articular injection of ovalbumin. After successful modeling, 30 New Zealand rabbits with arthritis were randomly divided into model control group, the high, medium and low dose groups of Tongbi capsule (90, 45, 22.5 mg·kg⁻¹) and prednisone group (5 mg·kg⁻¹). Another six normal rabbits were used as normal control group. After 24 hours of modeling, the rabbits in Tongbi capsule groups received intragastric (i.g.) administrations of Tongbi capsule at 90, 45, 22.5 mg·kg⁻¹·d⁻¹, and the rabbits of prednisone group received i.g. administrations of prednisone at 5 mg·kg⁻¹·d⁻¹ for 2 weeks. The rabbits in normal and model groups received the same volume of distilled water at the same time. The swelling degree of rabbit knee joint and local skin temperature were observed daily. After two weeks of administration, pathological changes of rabbit knee joint were examined by magnetic resonance imaging (MRI); the morphological changes of articular cartilage and synovial membrane were observed by microscope; and the contents of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in serum were detected by enzyme linked immunosorbent assay (ELISA).The results showed that 24 h after modeling, the knee joints of the rabbits were swollen, with red or dark redlocal skin, and fever, elevated local skin temperature and increased diameters of knee joints. Two weeks after modeling, the swelling of rabbit knee joints was obvious in model group; the joint cavities were filled with purulent fluid; joint synovial membranes were obviously thickened, and even joint cavities were fibrotic and cartilage surfaces showed slight defect; the surface of articular cartilage was obvious fibrosis; synovial epithelial cell proliferation was obvious and accompanied by extensive inflammatory cell infiltration; the levels of IL-1 and TNF-α were significantly higher as compared with those seen in model rabbits (P<0.05, P<0.01). After 1 and 2 weeks of administration, knee joint diameters and local skin temperatures were smaller or lower than thosein model group (P<0.05, P<0.01); The lesions of joint cartilage and synovial of all rabbits in each group were less than those in model group; IL-1 and TNF-α levels in serum were also lower than those in model group (P<0.05, P<0.01). The results reveal that high and medium doses of Tongbi capsule can suppress rheumatoid arthritis induced by ovalbumin in rabbits, reduce joint swelling, inhibit synovial epithelial and fiber hyperplasia and inflammatory cell infiltration, and alleviate articular cartilage damage. The mechanism may be associated with decreasing IL-1 and TNF-α levels in serum.

Effects of Tongbi capsule on joint lesions in rabbits with rheumatoid arthritis / 中国中药杂志

The purpose of this experiment is to observe the effects of Tongbi capsule on joint lesions in rabbit with rheumatoid arthritis induced by ovalbumin and explore the mechanism in order to provide reference for clinical application of Tongbi capsule. Rheumatoid arthritis in rabbits was induced by subcutaneous injection of emulsions of ovalbumin and Freund's complete adjuvant and intra articular injection of ovalbumin. After successful modeling, 30 New Zealand rabbits with arthritis were randomly divided into model control group, the high, medium and low dose groups of Tongbi capsule (90, 45, 22.5 mg·kg⁻¹) and prednisone group (5 mg·kg⁻¹). Another six normal rabbits were used as normal control group. After 24 hours of modeling, the rabbits in Tongbi capsule groups received intragastric (i.g.) administrations of Tongbi capsule at 90, 45, 22.5 mg·kg⁻¹·d⁻¹, and the rabbits of prednisone group received i.g. administrations of prednisone at 5 mg·kg⁻¹·d⁻¹ for 2 weeks. The rabbits in normal and model groups received the same volume of distilled water at the same time. The swelling degree of rabbit knee joint and local skin temperature were observed daily. After two weeks of administration, pathological changes of rabbit knee joint were examined by magnetic resonance imaging (MRI); the morphological changes of articular cartilage and synovial membrane were observed by microscope; and the contents of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in serum were detected by enzyme linked immunosorbent assay (ELISA).The results showed that 24 h after modeling, the knee joints of the rabbits were swollen, with red or dark redlocal skin, and fever, elevated local skin temperature and increased diameters of knee joints. Two weeks after modeling, the swelling of rabbit knee joints was obvious in model group; the joint cavities were filled with purulent fluid; joint synovial membranes were obviously thickened, and even joint cavities were fibrotic and cartilage surfaces showed slight defect; the surface of articular cartilage was obvious fibrosis; synovial epithelial cell proliferation was obvious and accompanied by extensive inflammatory cell infiltration; the levels of IL-1 and TNF-α were significantly higher as compared with those seen in model rabbits (<0.05, <0.01). After 1 and 2 weeks of administration, knee joint diameters and local skin temperatures were smaller or lower than thosein model group (<0.05, <0.01); The lesions of joint cartilage and synovial of all rabbits in each group were less than those in model group; IL-1 and TNF-α levels in serum were also lower than those in model group (<0.05, <0.01). The results reveal that high and medium doses of Tongbi capsule can suppress rheumatoid arthritis induced by ovalbumin in rabbits, reduce joint swelling, inhibit synovial epithelial and fiber hyperplasia and inflammatory cell infiltration, and alleviate articular cartilage damage. The mechanism may be associated with decreasing IL-1 and TNF-α levels in serum.