强骨通痹胶囊对大鼠骨关节炎炎症因子的影响及作用机制

Effect and Mechanism of Qianggu Tongbi Capsule on Osteoarthritis Factors in Rats

目的探讨强骨通痹胶囊对大鼠骨关节炎炎症因子的影响及作用机制。方法使用前交叉韧带切断法构建骨关节炎(osteoarthritis,OA)大鼠模型。将大鼠随机分为对照组、模型组、强骨通痹胶囊组和葡萄糖胺组。苏木精-伊红(HE)染色进行模型鉴定;甲苯胺蓝染色观察软骨组织变化;qRT-PCR检测大鼠关节炎软骨组织中Toll样受体4(TLR4)和核因子κB(NF-κB)mRNA的表达水平;酶联免疫(ELISA)试剂盒检测IL-1β、TNF-α和IL-6水平;免疫组化法检测TLR4、p-NF-κB水平。结果HE染色表明,与对照组相比,模型组膝关节内可见大量炎性细胞浸润,滑膜组织明显增厚,而强骨通痹胶囊和葡萄糖胺组的炎性细胞浸润和滑膜组织增厚不明显。甲苯胺蓝染色结果表明,与对照组相比,模型组软骨组织中的硫酸软骨素明显减少,而强骨通痹胶囊组和葡萄糖胺组的硫酸软骨素较模型组明显增加。与对照组相比,模型组的TLR4和NF-κB的mRNA表达水平和组织中蛋白含量以及组织IL-1β、TNF-α和IL-6水平均显著上升(均P<0.05);与模型组相比,强骨通痹胶囊组和葡萄糖胺组的TLR4和NF-κB的mRNA和蛋白表达水平以及组织IL-1β、TNF-α和IL-6水平均显著下降(均P<0.05)。结论强骨通痹胶囊能够通过抑制TLR4和NF-κB的表达,从而抑制炎症因子的产生,达到治疗骨关节炎的效果。

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 randomly 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 glucosamine 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 results of TLR4 and NF-κB,and the levels of IL-1β,TNF-αand 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-κB,and the levels of IL-1β,TNF-αand 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 achieve the effect of treatmenting osteoarthritis.