低强度脉冲聚焦超声通过上调PGAM5蛋白表达促进软骨细胞线粒体自噬  

Focused low-intensity pulsed ultrasound promotes chondrocyte mitophagy via up-regulating PGAM5 protein

作　　者：叶海霞[1] 虞乐华[1] 贾朗[1] YE Haixia;YU Lehua;JIA Lang(Department of Rehabilitation Medicine,the Second Affiliated Hospital of Chongqing Medical University,Chongqing,400010,China)

机构地区：[1]重庆医科大学附属第二医院康复医学科,重庆400010

出　　处：《第三军医大学学报》2021年第5期403-410,共8页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81802234)。

摘　　要：目的探讨低强度脉冲聚焦超声(focused low-intensity pulsed ultrasound,FLIPUS)对软骨细胞线粒体自噬活化的影响。方法提取C57BL/6J乳鼠膝关节原代软骨细胞进行体外实验,IL-1β处理细胞模拟骨关节炎样软骨细胞损伤并进行验证。细胞分为对照组、IL-1β组和IL-1β+FLIPUS组。RT-PCR检测各组Col2α1和MMP13 mRNA表达;Western blot检测各组Col2α1、MMP13、LC3B-Ⅱ、TOM20、TIM23、PGAM5、FUNDC1及p-FUNDC1(p-ser13)蛋白表达。结果成功提取原代软骨细胞并模拟骨关节炎样软骨细胞损伤。与对照组比较,IL-1β组Col2α1 mRNA和蛋白表达明显降低(P<0.01),MMP13 mRNA和蛋白表达明显增加(P<0.01);与IL-1β组相比,IL-1β+FLIPUS组Col2α1表达明显上升(P<0.05),MMP13表达明显下降(P<0.01)。与对照组相比,IL-1β组LC3B-Ⅱ蛋白表达显著降低(P<0.05);与IL-1β组相比,IL-1β+FLIPUS组LC3B-Ⅱ、PGAM5蛋白表达显著上升(P<0.01),TOM20、TIM23、p-ser13蛋白表达则明显下降(P<0.05)。结论FLIPUS可能通过上调软骨细胞PGAM5蛋白表达,使FUNDC1第13位丝氨酸去磷酸化,活化线粒体自噬。Objective To investigate the effect of focused low-intensity pulsed ultrasound(FLIPUS)on the activation of mitophagy in chondrocytes.Methods Chondrocytes were isolated from the knee articular cartilage of C57 BL/6 J suckling mice,and then primarily cultured.Then osteoarthritis-like chondrocyte injury was inflicted by IL-1βsimulation in the chondrocytes and identified.The chondrocytes were divided into 3 groups,that is,control,IL-1βand IL-1β+FLIPUS group.The mRNA expression of Col2α1 and MMP13 was detected by quantitative real-time PCR,and the protein levels of Col2α1,MMP13,LC3 B-Ⅱ,TOM20,TIM23,PGAM5,FUNDC1 and p-FUNDC1(p-ser13)were detected by Western blotting.Results We cultured primary chondrocytes and established osteoarthritis-like chondrocyte injury model successfully.Compared with the control group,the mRNA and protein levels of Col2α1 were significantly decreased(P<0.01),while the levels of MMP13 were significantly increased(P<0.01)in the IL-1βgroup.The levels of Col2α1 were obviously enhanced(P<0.05),while those of MMP13 were reduced(P<0.01)in the IL-1β+FLIPUS group than the IL-1βgroup.Compared with the control group,the protein expression of LC3 B-Ⅱwas remarkably reduced in the IL-1βgroup(P<0.05).The protein levels of LC3 B-Ⅱand PGAM5 were significantly increased(P<0.01),while those of TOM20,TIM23 and p-Ser13 were notably decreased(P<0.05)in the IL-1β+FLIPUS group than the IL-1βgroup.Conclusion FLIPUS may activate the mitophagy in chondrocytes though dephosphorylation of FUNDC1 Ser13 via up-regulating PGAM5 expression.

关 键 词：低强度脉冲聚焦超声 膝骨关节炎 线粒体自噬 PGAM5 FUNDC1 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]