肩袖损伤患者肌腱和骨组织中氧化应激状态及Beclin1、mTOR的表达分析

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摘要目的观察肩袖损伤发生时肌腱和肩袖止点足印区骨组织中氧化应激反应及Beclin1、雷帕霉素靶蛋白(mammalian target of rapamycin，mTOR)的差异表达情况。方法选取20例肩袖损伤患者，男10例，女10例；年龄30~75岁，平均(61.52±7.32)岁。全部患者均行肩关节镜下肩袖锚钉修补术，术中提取患者损伤的肌腱组织和肩袖止点足印区骨组织样本为损伤组，取正常的肌腱组织和足印区旁骨组织样本为对照组。使用2'，7'二氯二氢荧光素二乙酸酯(2'，7'dichlorodihydrofluorescein diacetate，DCFH-DA)法检测肌腱和骨组织样本中的总活性氧(reactive oxygen species，ROS)水平，二氢乙锭(dihydroethidium，DHE)法检测超氧化物歧化酶(superoxide dismutase，SOD)含量；逆转录聚合酶链式反应(reverse transcriotion-polymerase chain reaction，RT-PCR)检测肌腱和骨组织中Beclin1、mTOR mRNA的表达情况，免疫蛋白印迹(western blot，WB)检测Beclin1、p-mTOR/mTOR蛋白表达。结果与对照组比较，损伤组肌腱和骨组织中总ROS、Beclin1基因和蛋白上调(P＜0.05)，mTOR基因、p-mTOR蛋白下调(P＜0.05)，SOD在肌腱中下调、在骨组织中上调(P＜0.05)；损伤组的mTOR蛋白上调不明显(P＞0.05)。结论肩袖损伤时激发了肌腱和肩袖止点足印区骨组织中的氧化应激反应，同时可调节自噬因子Beclin1、mTOR的表达。

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	刘治军
	刘少津
	郑维蓬
	魏合伟
	廖志浩
	陈胜

关键词 ：肩袖损伤, 氧化应激, 自噬

Abstract：Objective Tobserve the oxidative stress response and the differential expression of Beclin1 and mTOR in the bone tissues of the tendon and the footprint area of the rotator cuff when the rotator cuff injury occurs.Methods Twenty patients with rotator cuff injury were selected，including 10 males and 10 females.The average age was(61.52±7.32).All patients underwent shoulder arthroscopic rotator cuff anchor repair.The injured tendon tissue and bone tissue samples at rotator cuff insertion were taken as the injured group，and the normal tendon tissue and bone tissue samples beside the footmark were taken as the control group.The DCFHDA method was used to detect the total reactive oxygen species(ROS) level in the tendon and bone tissue samples，the DHE method was used to detect the superoxide dismutase(SOD) content；the RTPCR technique was used to detect the expression of Beclin1 and mTOR mRNA in the tendon and bone tissue.Western blot(WB) detects Beclin1，p-mTOR/mTOR protein expression.Results Compared with the control group，in the injured group，total reactive oxygen species(ROS)，Beclin1 gene and protein were up-regulated(P＜0.05)，mTOR gene and p-mTOR protein were down-regulated(P＜0.05)，and superoxide dismutase(SOD)was down-regulated in the tendon and up-regulated in the bone tissue(P＜0.05).mTOR protein in the injury group was not significantly up-regulated(P＞0.05).Conclusion Rotator cuff injury stimulates the oxidative stress response in the tendon and in the bone tissue of rotator cuff stop point and foot print area，and can also regulate the expression of autophagy factors Beclin1 and mTOR.It affects the healing of the rotator cuff，indicating oxidative stress and autophagy，and has an important regulatory effect on tendonbone healing.

基金资助:广东省医学科学技术研究基金项目(B2020030)
广东省中医药局中医药科研项目(20203011)

通讯作者: 魏合伟

作者简介: 刘治军(1982－ )，男，主治医师，广州中医药大学第三附属医院运动医学科，doctoransonliu@qq.com

引用本文:

刘治军，刘少津，郑维蓬，魏合伟，廖志浩，陈胜. 肩袖损伤患者肌腱和骨组织中氧化应激状态及Beclin1、mTOR的表达分析[J]. 实用骨科杂志, 2022, 28(12): 1079-.

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http://www.sygkzz.com/CN/ 或 http://www.sygkzz.com/CN/Y2022/V28/I12/1079

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