辛伐他汀对膝骨关节炎大鼠MMP-3、MMP-9、TGF-β1表达的影响及作用机制

Effect of simvastatin on expression levels of MMP-3,MMP-9 and TGF-β1 of rats with osteoarthritis of the knee

目的探讨辛伐他汀对膝骨关节炎大鼠基质金属蛋白酶(MMP)-3、MMP-9、转化生长因子(TGF)-β1表达水平的影响及其作用机制。方法 40只胶原诱导膝骨关节炎造模成功的大鼠随机分为模型组和低剂量组、中剂量组和高剂量组,另选取10只未造模大鼠作为对照组。分别给予低、中、高剂量组0.01、0.1、0.5 mg/ml辛伐他汀稀释液灌胃,剂量为0.2 ml/kg,同时给予对照组和模型组同等剂量无菌生理盐水灌胃。6 w后处死,采用免疫组化法检测组织中Caspase-3表达情况。对比5组给药前、3 w和6 w后血清MMP-3、MMP-9和TGF-β1水平变化及Caspase-3表达情况。结果用药前对照组血清MMP-3、MMP-9和TGF-β1表达水平均较其他各组显著降低(P<0.05),3 w后和6 w后对照组和模型组血清因子水平均无显著变化(P>0.05),而低、中、高剂量组均随着用药时间延长而显著降低(P<0.05),3 w和6 w后各项指标水平按照由高到低排列为模型组、低剂量组、中剂量组、高剂量组和对照组,且每两组间差异均有统计学意义(P<0.05);高剂量组Caspase-3 OD值远远高于中剂量组和低剂量组(P<0.05),中剂量组远远高于低剂量组(P<0.05),低剂量组较对照组和模型组高(P<0.05),对照组和模型组无显著差异(P>0.05)。结论辛伐他汀可改善膝骨关节炎大鼠血清MMP-3、MMP-9和TGF-β1水平,在一定范围内剂量越高作用效果越好,推测可能和Caspase-3异常激活有关。

Objective To discuss the effect of simvastatin on expression levels of matrix metalloproteinase （MMP） -3, MMP-9 and transforming growth factor （TGF） -β1 of rats with osteoarthritis of the knee, and to explore the mechanism of action. Methods 40 successful modeling rats with osteoarthritis of the knee induced by collagen were randomly divided into model, low dose, middle dose and high dose groups, and another 10 rats without modeling were selected as the control group. The low, middle and high dose groups were respectively given 0. 01,0.1,0. 5mg/ml of simvastatin diluted solution, with the dose of 0.2 ml/kg, then the control and model groups were given same dose of sterile saline to fill the stomach at the same time. 6 weeks later, the expression of Caspase-3 was detected by immunohistochemistry. The changes of serum levels of MMP-3, MMP-9, and TGF-IM before medication,after 3 and 6 weeks were compared among the 5 groups, as well as the expressions of Caspase-3, then the mechanism of action of simvastatin on rats with osteoarthritis of the knee was analyzed. Results The expression levels of serum MMP-3, MMP-9, and TGF-β1 in the control group before medication were significantly lower than that in the other groups （P〈0.05）, and the levds in the low, middle and high dose groups were significantly decreased with the increase of medication time （P〈0.05）, then the levels after 3 weeks and 6 weeks were ranked from high to low as follows：the model group,low dose group, middle dose group, high dose group and control group （ P〈0. 05 ）. The OD value of Caspase-3 in the high dose group was far higher than that in the middle and low dose groups （ P〈0.05 ）, while the value in the middle dose group was far higher than that in the low dose group （ P〈0.05 ）, as well as the value in the low dose group was far higher than that in the control group and model group （P〈0.05）, but there was no signifi- cant difference between the control group and model group （P〉0.05）. Conclusions Simvastatin can improve the serum levels of MMP-3, MMP-9, and TGF-β1 in rats with osteoarthritis of the knee, and in a certain range the higher the dose, the better the effect, and it can be speculated that it may be related to the abnormal activation of Caspase-3.