体外冲击波干预骨关节炎大鼠软骨细胞的增殖和自噬  

Effects of extracorporeal shock wave on the proliferation and autophagy of chondrocytes from osteoarthritis rats

作　　者：王欢欢 王青 唐鹏 张睿 闵红巍[1,3] Wang Huanhuan;Wang Qing;Tang Peng;Zhang Rui;Min Hongwei(China Rehabilitation Research Center,School of Rehabilitation,Capital Medical University,Beijing 100068,China;Chinese Institute of Rehabilitation Science,Beijing 100068,China;Joint Disease Treatment Center,Beijing Bo’ai Hospital,China Rehabilitation Research Center,Beijing 100068,China)

机构地区：[1]首都医科大学康复医学院,中国康复研究中心,北京市100068 [2]中国康复科学所,北京市100068 [3]中国康复研究中心北京博爱医院关节病诊疗中心,北京市100068

出　　处：《中国组织工程研究》2023年第2期252-257,共6页Chinese Journal of Tissue Engineering Research

基　　金：中央级公益性科研院所基本科研业务费专项基金项目(2019CZ-12),项目负责人:王青。

摘　　要：背景:体外冲击波是一种新兴的治疗方法,被广泛应用于各个医疗领域,对骨关节炎的发生发展具有重要作用。目的:通过研究体外冲击波对大鼠骨关节炎软骨细胞增殖和自噬的影响,探讨体外冲击波在治疗骨性关节炎过程中的作用机制。方法:选取清洁级1周龄SD大鼠,获取四肢关节软骨进行原代软骨细胞的分离与培养。将第3代软骨细胞按随机数表法随机分为对照组(未加任何处理)、白细胞介素1β组(10μg/L白细胞介素1β刺激24 h)、体外冲击波组(1.5×10^(5)Pa体外冲击波干预500次)、白细胞介素1β+体外冲击波组(10μg/L白细胞介素1β刺激24 h后1.5×10^(5)Pa体外冲击波干预500次)。CCK-8法检测各组软骨细胞活力和增殖情况,实时荧光定量PCR法检测各组软骨细胞自噬基因Beclin 1、P62、Atg5和LC3 mRNA的表达。结果与结论:①与对照组相比,白细胞介素1β组软骨细胞活力和增殖能力下降(P<0.05);与白细胞介素1β组相比,白细胞介素1β+体外冲击波组软骨细胞活力和增殖能力有所提高(P<0.05);②与对照组相比,白细胞介素1β组自噬基因Beclin 1、Atg5和LC3的表达下降,P62的表达增加(P<0.05);与白细胞介素1β组相比,白细胞介素1β+体外冲击波组自噬基因Beclin 1、Atg5和LC3的表达显著增加,P62表达下降(P<0.05);③结果表明,体外冲击波可促进骨关节炎软骨细胞增殖,并提高骨关节炎软骨细胞的自噬水平,从而参与调节骨关节炎的进展。BACKGROUND:Extracorporeal shock wave is a new treatment method that has been widely used in various medical fields and plays an important role in the occurrence and development of osteoarthritis.OBJECTIVE:To investigate the effects of extracorporeal shock wave on the proliferation and autophagy of chondrocytes from osteoarthritis rats in vitro and to explore the mechanism of extracorporeal shock wave in the treatment of osteoarthritis.METHODS:Primary chondrocytes were isolated and cultured from the articular cartilage of extremities of 1-week-old clean-grade Sprague-Dawley rats.Chondrocytes at passage 3 were randomly divided into control group(without any treatment),interleukin-1βgroup(10μg/L interleukin-1βfor 24 hours),extracorporeal shock wave group(1.5×10^(5)Pa extracorporeal shock wave for 500 times),interleukin-1β+extracorporeal shock wave group(10μg/L interleukin-1βfor 24 hours+1.5×10^(5)Pa extracorporeal shock wave for 500 times).The viability and proliferation of chondrocytes were detected by cell counting kit-8 method.The mRNA expressions of Beclin 1,P62,Atg5 and LC3 were detected by real-time fluorescence quantitative PCR.RESULTS AND CONCLUSION:Stimulation with interleukin-1βfor 24 hours decreased the viability and proliferation of chondrocytes compared with the control group(P<0.05).Compared with the interleukin-1βgroup,chondrocyte viability and proliferation were increased in the interleukin-1β+extracorporeal shock wave group(P<0.05).The expression of Beclin 1,Atg5 and LC3 was down-regulated and P62 expression was increased in the interleukin-1βgroup compared with the control group(P<0.05),whilst the expression of Beclin 1,Atg5 and LC3 was significantly increased and the expression of P62 was decreased in the interleukin-1β+extracorporeal shock wave group compared with the interleukin-1βgroup(P<0.05).These results suggest that extracorporeal shock wave can promote the proliferation and autophagy of osteoarthritic chondrocytes,thus regulating the progression of osteoarthritis.

关 键 词：体外冲击波 骨关节炎 软骨细胞 增殖 自噬 大鼠 

分 类 号：R459.9[医药卫生—治疗学] R-33[医药卫生—临床医学]