针刀调控软骨细胞自噬维持骨关节炎软骨细胞的稳态  

Regulation of chondrocyte autophagy by acupotomy to promote chondrocyte homeostasis in osteoarthritis

作　　者：贾小飞 冉丽 马小双 黑晓燕 刘佳妮 杨楠 马海滨[1] 常敬鹏 Jia Xiaofei;Ran Li;Ma Xiaoshuang;Hei Xiaoyan;Liu Jiani;Yang Nan;Ma Haibin;Chang Jingpeng(Rehabilitation Medicine Center,People’s Hospital of Ningxia Hui Autonomous Region(People’s Hospital of Ningxia Hui Autonomous Region,Ningxia Medical University),Yinchuan 750001,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏回族自治区人民医院(宁夏医科大学附属自治区人民医院)康复医学科,宁夏回族自治区银川市750001

出　　处：《中国组织工程研究》2024年第34期5452-5457,共6页Chinese Journal of Tissue Engineering Research

基　　金：宁夏重点研发项目(引才专项)(2020BEB04045),项目负责人:贾小飞。

摘　　要：背景:针刀是治疗骨关节炎的有效方法,临床疗效肯定,但具体机制尚未明确。目的:探讨针刀调控软骨细胞自噬维持骨关节炎软骨细胞稳态的作用。方法:采用随机数字表法将28只新西兰兔分为对照组、骨关节炎组、针刀组和透明质酸组,每组7只,后3组保持左后肢过伸位采用石膏绷带固定6周建立膝骨关节炎模型,建模成功后,对照组与骨关节炎组不进行任何治疗,针刀组给予针刀治疗(15 min/次,1次/周),透明质酸组膝关节腔内注射透明质酸钠溶液(1次/周),均连续治疗5周。末次治疗后第2天,采用DR片观察兔膝关节宏观结构,透射电镜观察软骨细胞超微结构,Tunel染色检测软骨细胞凋亡,Western blot检测PI3K/Akt/mTOR通路及自噬相关蛋白表达。结果与结论:(1)DR片显示,骨关节炎组兔膝关节间隙变窄、周围骨赘形成;相较于骨关节炎组,透明质酸组、针刀组兔膝关节间隙增宽,周围骨赘减少;(2)透射电镜下可见骨关节炎组兔膝关节软骨细胞内自噬体数量减少,细胞核皱缩、核膜破裂,细胞器形态不完整、细胞呈现明显死亡趋向;相较于骨关节炎组,透明质酸组与针刀组自噬体明显增加,细胞核核膜完整,细胞生存状态良好;(3)Tunel染色结果显示,相较于骨关节炎组,透明质酸组、针刀组凋亡软骨细胞明显减少;(4)Western blot检测结果显示,与对照组比较,骨关节炎组Beclin1、Cath D和LC3Ⅱ/LC3Ⅰ蛋白表达降低(P <0.05),p-Akt/Akt、p-mTOR/mTOR蛋白表达升高(P <0.05);与骨关节炎组比较,透明质酸组和针刀组Beclin1、Cath D和LC3Ⅱ/LC3Ⅰ蛋白表达升高(P <0.05),p-Akt/Akt、p-mTOR/mTOR蛋白表达降低(P <0.05);(5)结果表明,针刀可通过抑制PI3K/Akt/mTOR信号通路传导来提高软骨细胞自噬水平,维持软骨细胞稳态,最终延缓软骨退变。BACKGROUND:Acupotomy is an effective method for the clinical treatment of osteoarthritis,with affirmed clinical outcomes,but the specific mechanisms remain unclear OBJECTIVE:To investigate the role of acupotomy in modulating chondrocyte autophagy to promote chondrocyte homeostasis in osteoarthritis.METHODS:Twenty-eight New Zealand rabbits were randomly divided into control group,osteoarthritis group,acupotomy group,and hyaluronic acid group,with seven rabbits in each group.The knee osteoarthritis rabbit model was prepared using the Videman method in the latter three groups.After modeling,the control group and osteoarthritis group received no interventions.The acupotomy group received acupotomy treatment 15 minutes per time,once a week,while the hyaluronic acid group received intra-articular injection of hyaluronic acid once a week,with a continuous treatment duration of 5 weeks.The day after the final intervention,knee joint macrostructure was observed using DR imaging,chondrocyte ultrastructure was examined through transmission electron microscopy,apoptosis of chondrocytes was assessed using Tunel staining,and western blot analysis was used to detect the expression of proteins related to the PI3K/Akt/mTOR pathway.RESULTS AND CONCLUSION:The DR imaging results revealed that the osteoarthritis group exhibited narrowed knee joint spaces and the formation of periarticular osteophytes,while the hyaluronic acid group and acupotomy group showed widened knee joint spaces with a reduction in periarticular osteophytes.Transmission electron microscopy results demonstrated a decreased number of autophagosomes in chondrocytes in the osteoarthritis group,along with nuclear shrinkage,nuclear membrane rupture,incomplete organelle morphology,and a clear tendency towards cell death.In contrast,both the hyaluronic acid group and acupotomy group exhibited a significant increase in autophagosomes,intact nuclear membranes,and a well-preserved cellular state.Tunel staining results indicated a considerable decrease in the number of apop

关 键 词：针刀 软骨细胞 自噬 骨关节炎 软骨细胞稳态 透明质酸 

分 类 号：R493[医药卫生—康复医学] R318[医药卫生—临床医学] R244.9