川芎嗪对膝骨性关节炎大鼠软骨BMP-2、Smad1及BMP-2mRNA、Smad1mRNA表达的影响

摘要

目的 观察川芎嗪治疗膝骨关节炎(knee osteoarthritis, KOA)大鼠模型关节软骨中骨形态发生蛋白-2 (bone morphogenetic protein-2,BMP-2)、Smad1表达的变化,并探讨川芎嗪治疗KOA的作用机制.方法 制作大鼠膝关节炎模型,将建模成功的大鼠纳入模型对照组,常规饲养,川芎嗪高剂量组、川芎嗪低剂量组、阳性对照组行灌,正常组和模型组行等量生理盐水治疗6 w,实验结束后,各组大鼠分别行HE切片软骨Mankin评分、Western Blot 检测软骨组织BMP-2、Smad1及实时荧光定量PCR检查BMP-2mRNA、Smad1mRNA的变化.结果 各组软骨Mankin评分中,川芎嗪各剂量组和阳性对照组均较空白组高(P＜0.05),均较模型组低(P＜0.05),且川芎嗪高剂量组＜阳性对照组＜川芎嗪低剂量组(P＜0.05).正常组、川芎嗪各剂量组及阳性对照组中软骨BMP-2、Smad1及BMP-2mRNA、Smad1mRNA相对表达量较模型组均高于模型组,差异有统计学意义(P＜0.05);与阳性对照组相比,川芎嗪高剂量组BMP-2、Smad1及BMP-2mRNA、Smad1mRNA表达量明显高于阳性对照组(P＜0.05),而川芎嗪低剂量组其表达则均低于阳性对照组.结论 川芎嗪可能通过上调早期KOA大鼠BMP-2mRNA及Smad1mRNA基因的表达,从而促进BMP-2及Smad1蛋白的表达,这可能是在某种程度上减轻关节软骨退变、修复软骨损伤作用的机制之一.%Objective To observe the effect of Ligustrazine on the expression of bone morphogenetic protein-2 (BMP-2) and Smad1 in the articular cartilage of knee osteoarthritis rats (KOA), and to explore the mechanism of Ligustrazine in the treatment of KOA.Methods Establishing knee arthritis rat model, and the successfully established KOA model rats were selected and divided randomly into model control group, routine feeding group, high dose Ligustrazine group, low dose Ligustrazine group and positive control group.The normal group and the model group were treated with the same amount of physiological saline.At the end of the experiment, Mankin score was performed on HE slices of cartilage in each group.BMP-2 and Smad1 were detected by Western Blot and real-time quantitative PCR were used to detect the changes of BMP-2 mRNA and Smad1 mRNA in cartilage tissue.Results For Mankin score, the two dose groups of Ligustrazine and the positive control group had higher score than the blank group (P＜0.05), but lower score than the model group (P＜0.05), with the score of the high dose Ligustrazine group significantly greater than the positive control group, and the positive control group significantly greater than the low dose group (P ＜0.05).The expression of BMP-2, Smad1, BMP-2mRNA and Smad1mRNA in the cartilage of normal group and Ligustrazine groups were higher than that in the model group, the differences were statistically significant (P＜0.05).Compared with the positive control group, the expression of BMP-2, Smad1, BMP-2mRNA and Smad1mRNA in the high dose Ligustrazine group was significantly higher (P＜0.05), while the expression in the low dose Ligustrazine group was significantly lower.Conclusion Ligustrazine may promote the expression of BMP-2 and Smad1 protein by up-regulating the expression of BMP-2mRNA and Smad1mRNA gene in early KOA rats, which may be a mechanism to alleviate cartilage degeneration and repair cartilage injury to some extent.