动、静态应力刺激在大鼠跟腱损伤早期愈合过程中抗粘连作用及其机制的研究  

Study on anti-adhesion effect and mechanism of dynamic and static stress stimulation during early healing process of rat Achilles tendon injury

作　　者：吴佳妮 蒋滢梓 汪官玉 王礼燎 鲍捷[2] 王骏 WU Jiani;JIANG Yingzi;WANG Guanyu;WANG Liliao;BAO Jie;WANG Jun(Department of Rehabilitation Medicine,Wuxi Ninth People’s Hospital,Wuxi Jiangsu,214000,P.R.China;School of Physical Education,Soochow University,Suzhou Jiangsu,215000,P.R.China)

机构地区：[1]无锡市第九人民医院康复医学科,江苏无锡214000 [2]苏州大学体育学院,江苏苏州215000

出　　处：《中国修复重建外科杂志》2024年第11期1391-1398,共8页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：2021年度无锡市“太湖人才计划”。

摘　　要：目的探讨动、静态应力刺激对大鼠跟腱损伤早期愈合过程中的抗粘连作用及其机制。方法取15只4~6周龄雄性SD大鼠跟腱组织,采用酶消化法分离培养跟腱细胞;通过TNF-α处理大鼠跟腱细胞,构建损伤愈合早期跟腱细胞模型,分别施加动态应力刺激(动态组)和静态应力刺激(静态组),对照组未作处理。采用活/死细胞双染色检测细胞活性,ELISA法检测α平滑肌肌动蛋白(αsmooth muscle actin,α-SMA)表达,实时荧光定量PCR检测Ⅰ型胶原蛋白(collagen typeⅠ,COL1A1)、Ⅲ型胶原蛋白(collagen typeⅢ,COL3A1)和Scleraxis(SCX)的mRNA表达。对30只4~6周龄雄性SD大鼠施行跟腱断裂缝合术,随机分为动态组(动态应力刺激)、静态组(静态应力刺激)和对照组(未作处理),每组10只。术后8 d取材,进行HE染色及评分评价跟腱愈合情况,免疫组织化学染色检测COL1A1和COL3A1蛋白表达,Western blot检测α-SMA和SCX蛋白表达,生物力学拉伸测试检测最大肌腱断裂力和肌腱刚度。结果在体外细胞实验中,动态组较静态组活细胞数量明显增加,α-SMA蛋白表达下降,COL3A1 mRNA相对表达量下降、SCX mRNA相对表达量上升,差异均有统计学意义(P<0.05)。在体内动物实验中,动态组较静态组肌腱愈合情况更好,HE染色评分更低,COL1A1蛋白表达上升、COL3A1蛋白表达下降,SCX蛋白相对表达量上升、α-SMA蛋白相对表达量下降,肌腱刚度上升,差异均有统计学意义(P<0.05)。结论与静态应力刺激相比,动态应力刺激改善了大鼠跟腱瘢痕组织纤维化,促进了跟腱生物力学性能恢复,具有明显抗粘连作用。Objective To investigate the anti-adhesive effect and underlying mechanism of dynamic and static stress stimulation on the early healing process of rat Achilles tendon injury.Methods Achilles tendon tissues of 15 male Sprague Dawley(SD)rats aged 4-6 weeks were isolated and cultured by enzyme digestion method.Rat Achilles tendon cells were treated with tumor necrosis factorαto construct the Achilles tendon injury cell model,and dynamic stress stimulation(dynamic group)and static stress stimulation(static group)were applied respectively,while the control group was not treated.Live/dead cell double staining was used to detect cell activity,ELISA assay was used to detect the expression ofαsmooth muscle actin(α-SMA),and real-time fluorescence quantitative PCR was used to detect the mRNA expression of collagen typeⅠ(COL1A1),collagen typeⅢ(COL3A1),and Scleraxis(SCX).Thirty male SD rats aged 4-6 weeks underwent Achilles tendon suture and were randomly divided into dynamic group(treated by dynamic stress stimulation),static group(treated by static stress stimulation),and control group(untreated),with 10 rats in each group.HE staining and scoring were performed to evaluate the healing of Achilles tendon at 8 days after operation.COL1A1 and COL3A1 protein expressions were detected by immunohistochemical staining,α-SMA and SCX protein expressions were detected by Western blot,and maximum tendon breaking force and tendon stiffness were detected by biomechanical stretching test.Results In vitro cell experiment,when compared to the static group,the number of living cells in the dynamic group was higher,the expression ofα-SMA protein was decreased,the relative expression of COL3A1 mRNA was decreased,and the relative expression of SCX mRNA was increased,and the differences were all significant(P<0.05).In the in vivo animal experiment,when compared to the static group,the tendon healing in the dynamic group was better,the HE staining score was lower,the expression of COL1A1 protein was increased,the expression of COL3A1 p

关 键 词：动、静态应力刺激 跟腱细胞 大鼠 愈合早期 抗粘连 

分 类 号：R686.1[医药卫生—骨科学]