力学刺激通过整合素α5β1介导半月板纤维软骨细胞的凋亡  

Mechanical stimulation mediates apoptosis of meniscal fibrochondrocytes via integrin α5β1

作　　者：杨化群 付岳松 王法正[1] YANG Huaqun;FU Yuesong;WANG Fazheng(Department of Sports Medicine,First People’s Hospital of Kashi Area,Kashi 844000,China)

机构地区：[1]喀什地区第一人民医院运动医学科

出　　处：《山西医科大学学报》2019年第8期1134-1139,共6页Journal of Shanxi Medical University

基　　金：新疆维吾尔自治区自然科学基金资助项目(2016D01C021)

摘　　要：目的研究力学刺激对半月板纤维软骨细胞(meniscal fibrochondrocytes,MFs)凋亡的影响,并探讨可能的分子机制。方法通过酶消化法从大鼠半月板组织中分离MFs,实验首先分为两组:对照组(无力学刺激)和力学刺激组(10%牵张力,0.5 Hz,8 h/d),通过力学刺激仪器诱导MFs损伤细胞模型,CCK-8法和流式细胞术分别检测力学刺激对MFs活力和凋亡的影响。然后将实验分为四组:对照组(无力学刺激)、力学刺激组(10%牵张力,0.5 Hz,8 h/d)、对照siRNA(NC-siRNA)力学刺激组和干扰α5β1(α5β1 siRNA)力学刺激组,其中α5β1 siRNA和NC-siRNA力学刺激组细胞通过lipofectamine 2000将α5β1 siRNA和对照siRNA转染到MFs,然后进行力学刺激;CCK-8法和流式细胞术分别检测α5β1 siRNA和力学刺激对MFs活力和凋亡的影响;Western blot检测力学刺激和α5β1 siRNA对MFs Bcl2和cleaved caspase3蛋白表达的影响。结果CCK-8结果显示,与对照组细胞活力比较,力学刺激组MFs细胞活力显著降低;与NC-siRNA力学刺激组相比,α5β1 siRNA力学刺激组细胞活力显著升高,差异具有统计学意义(P<0.01)。流式检测结果显示,与对照组比较,力学刺激组MFs细胞凋亡率明显升高;与NC-siRNA力学刺激组相比,α5β1 siRNA力学刺激组细胞凋亡率显著降低,差异具有统计学意义(P<0.01)。Western blot结果显示,与对照组比较,力学刺激组MFs细胞Bcl2表达显著降低,α5β1和cleaved caspase3表达明显升高;而与NC-siRNA力学刺激组相比,α5β1 siRNA力学刺激组细胞Bcl2表达显著升高,α5β1和cleaved caspase3表达明显降低,差异具有统计学意义(P<0.01)。结论力学刺激可能通过促进整合素α5β1表达,抑制Bcl2表达,促进cleaved caspase3表达,激活线粒体凋亡通路,促进MFs凋亡。Objective To study the effects of mechanical stimulation on apoptosis of meniscal fibrochondrocytes(MFs)and to explore its possible molecular mechanism.Methods The MFs were isolated from rat meniscus tissue by enzymatic digestion.First,the experiment was divided into two groups:control group(no mechanical stimulation)and mechanical stimulation group(10%tension,0.5 Hz,8 h/d).CCK-8 method and flow cytometry were used to detect the effects of mechanical stimulation on MFs activity and apoptosis.Then,the experiment was divided into four groups:control group(no mechanical stimulation),mechanical stimulation group(10%tension,0.5 Hz,8 h/d),negative control siRNA(NC-siRNA)mechanical stimulation group andα5β1(α5β1 siRNA)mechanical stimulation group.Inα5β1 siRNA group and NC-siRNA mechanical stimulation group,α5β1 siRNA and NC-siRNA were transfected into MFs cells by lipofectamine 2000.The effects of mechanical stimulation andα5β1 siRNA on the proliferation and apoptosis of MFs were detected by CCK8 and flow cytometry.The effects of mechanical stimulation andα5β1 siRNA on the expression of Bcl2 and cleaved caspase3 proteins were detected by Western blotting.Results The results of CCK8 showed that compared with control group,the cell viability of the MFs was significantly decreased in mechanical stimulation group(P<0.01),while compared with NC-siRNA mechanical stimulation group,the cell viability was significantly increased inα5β1 siRNA mechanical stimulation group(P<0.01).Flow cytometry results showed that the apoptosis rate of MFs cells in mechanical stimulation group was significantly higher than that in control group(P<0.01),while the apoptosis rate inα5β1 siRNA mechanical stimulation group was significantly lower than that of NC-siRNA mechanical stimulation group(P<0.01).Western blotting results showed that compared with control group,the expression of Bcl2 in MFs cells was significantly decreased and the expression ofα5β1 and cleaved caspase3 was significantly increased in mechanical stimulation grou

关 键 词：力学刺激 半月板纤维软骨细胞 整合素Α5Β1 细胞凋亡 

分 类 号：R363[医药卫生—病理学]