青藤碱对兔膝骨关节炎模型软骨Toll样受体2、4及髓样分化因子88表达的影响

Effects of Sinomenine on Expressions of TLR-2, TLR-4 and MyD88 in Cartilage of Rabbit Knee Osteoarthritis Model

目的观察不同剂量青藤碱对兔膝骨关节炎模型关节软骨中Toll样受体(TLR)2、4和髓样分化因子88(My D88)表达的影响,探讨其相关作用机制。方法采用Hulth法建立膝骨关节炎模型。实验兔随机分为空白组、模型组和青藤碱低、中、高剂量组,空白组不予任何处理,模型组和青藤碱低、中、高剂量组膝关节腔分别注射生理盐水和0.2 mL(5 mg)、0.35 mL(8.75 mg)、0.5 mL(12.5 mg)青藤碱,共干预10次。实时荧光定量PCR和Western blot检测关节软骨TLR2、TLR4和My D88的mRNA和蛋白表达。结果模型组TLR2、TLR4、My D88的mRNA和蛋白表达较空白组均显著升高(P<0.01);与模型组比较,青藤碱中、高剂量组TLR2、TLR4、My D88的mRNA和蛋白表达均显著下调(P<0.05,P<0.01)。结论青藤碱可通过下调TLR/My D88通路中关键分子的表达抑制兔膝骨性关节炎软骨免疫反应。

Objective To observe the effects of different doses of sinomenine on toll-like receptor（TLR）2, TLR4 and myeloid differentiation factor 88（Myd88） in cartilage of rabbit knee osteoarthritis model; To discuss its mechanism of action. Methods Knee osteoarthritis model was established by Hulth method. Experimental rabbits were randomly divided into control group, model group, sinomenine low-, medium-and high-dose groups. Blank group received no processing. Model group, sinomenine low-, medium-and high-dose groups received intra-articular injection of normal saline and sinomenine 0.2 mL（5 mg）, 0.35 mL（8.75 mg） and 0.5 mL（12.5 mg） respectively, for 10 times. Real-time fluorescence quantitative PCR and Western blot were used to detect m RNA and protein expressions of TLR2, TLR4 and My D88 in cartilage. Results m RNA and protein expressions of TLR2, TLR4 and MyD88 in model group significantly increased compared with control group（P0.01）; compared with model group, m RNA and protein expressions of TLR2, TLR4 and My D88 in sinomenine medium-and high-dose groups decreased significantly（P0.05, P0.01）. Conclusion Sinomenine can inhibit the cartilage immune response of rabbit knee osteoarthritis by down-regulating expressions of key molecules in the TLR/My D88 pathway.