吉非替尼对兔膝骨性关节炎的病理特征及相关因子的影响

Effects of gefitinib on pathological features of rabbit knee osteoarthritis and related factors

目的探究吉非替尼对兔膝骨性关节炎的病理特征及相关因子的影响。方法选取雌性新西兰大白兔60只,采用随机分组法将大白兔分为空白对照组、模型组、低剂量吉非替尼组和高剂量吉非替尼组各15只。除15只空白对照组,其他各组大白兔制成骨性关节炎(OA)模型,造模1周后,低剂量吉非替尼组使用4×10-1 mg/kg吉非替尼处理,高剂量吉非替尼组使用8×10-1 mg/kg吉非替尼处理。分别使用蛋白质印记法检测各组大白兔自噬蛋白Beclin1、LC3、p62表达情况;采用免疫荧光检测LC3含量;使用Elisa试剂盒检测大白兔相关炎症因子IL-1β、Il-6、TNF-α水平;检测软骨降解酶MMP-13及Adamts5表达水平;HE染色观察各组大白兔软骨形态。结果相比空白对照组,模型组Beclin1、LC3、p62水平无显著变化(P>0.05),IL-1β、Il-6、TNF-α水平显著升高,MMP-13及Adamts5表达水平显著降低(P>0.05);相比模型组,低剂量吉非替尼组Beclin1、LC3、MMP-13及Adamts5表达水平显著升高,p62、IL-1β、Il-6、TNF-α水平显著降低(P>0.05);相比低剂量吉非替尼组,高剂量吉非替尼组Beclin1、LC3、MMP-13及Adamts5表达水平显著升高,p62、IL-1β、Il-6、TNF-α水平显著降低(P>0.05)。结论吉非替尼可有效治疗骨性关节炎,且在一定剂量范围内呈剂量依赖性,其作用机制与提高自噬能力、降低炎症反应相关。

Objective To explore effects of gefitinib on pathological features of rabbit knee osteoarthritis(KOA)and related factors.Methods 60 female New Zealand white rabbits were enrolled.They were randomly divided into blank control group,model group,low-dose gefitinib group and high-dose gefitinib group,15 cases in each group.Except blank control group,white rabbits in the other groups were made into osteoarthritis(OA)models.At 1 week after modeling,low-dose gefitinib group and high-dose gefitinib group were treated with 4×10-1 mg/kg and 8×10-1 mg/kg gefitinib,respectively.The expression of autophagy proteins Beclin1,LC3 and p62 in each group were detected by Western blotting.The content of LC3 was detected by immunofluorescence.The levels of IL-1β,Il-6 and TNF-αwere detected by Elisa kit.The expression levels of cartilage degrading enzymes MMP-13 and Adamts5 were detected.HE staining was performed to observe cartilage morphology of rabbits in each group.Results Compared with blank control group,there was no significant change in Beclin1,LC3 or p62 level in model group(P>0.05),levels of IL-1β,Il-6 and TNF-αwere significantly increased,and expression levels of MMP-13 and Adamts5 were significantly decreased(P>0.05).Compared with model group,expression levels of Beclin1,LC3,MMP-13 and Adamts5 were significantly increased in low-dose gefitinib group,while levels of p62,IL-1β,Il-6 and TNF-αwere significantly decreased(P>0.05).Compared with low-dose gefitinib group,expression levels of Beclin1,LC3,MMP-13 and Adamts5 were significantly increased in high-dose gefitinib group,while levels of p62,IL-1β,Il-6 and TNF-αwere significantly decreased(P>0.05).Conclusion Gefitinib can effectively treat OA,showing dose-dependence within certain dose range.And its mechanism of action is related to improving autophagy ability and reducing inflammatory response.