雷公藤甲素对同种异体肌腱移植修复鸡肌腱缺损的作用

EFFECT OF TRIPTOLIDE ON ALLOGENIC TENDON TRANSPLANTATION IN REPAIRING TENDON DEFECT IN CHICKEN

目的雷公藤甲素具有抗排斥作用,探讨其在同种异体肌腱移植修复鸡肌腱缺损中的作用。方法 4月龄健康清洁级雄性来亨鸡64只,体重1.9～2.3 kg,取右足第3足趾肌腱制备肌腱缺损模型并行同种异体肌腱修复。根据是否给予雷公藤甲素,随机分为两组(n=32)。实验组术后给予雷公藤甲素100μg/(kg.d),共3周;对照组正常喂养。术后观察动物一般情况,于1、2、3、4周各组取4只动物大体观察移植肌腱情况,其中1、3周取材行组织学观察、2、4周行透射电镜观察。术后3、6周各组另取8只动物采血行流式细胞学检测,取肌腱行生物力学检测。结果术后实验动物均存活至实验完成。标本大体观察显示,随时间延长,两组肌腱周围充血、水肿缓解,实验组见疏松粘连带,对照组为广泛致密纤维组织粘连带。组织学观察示,术后1、3周实验组炎性反应均较对照组轻。透射电镜观察示,术后2、4周,实验组成纤维细胞核大,常染色质丰富,异染色质较少;对照组成纤维细胞细胞质内有少量粗面内质网,腔小,内容物少。术后3、6周,实验组CD4+、CD8+T淋巴细胞均较对照组少,CD4+/CD8+T淋巴细胞比值低于对照组;实验组肌腱最大拉伸断裂强度大于对照组,拉断粘连带功耗小于对照组;以上指标比较差异均有统计学意义(P<0.05)。结论雷公藤甲素可以降低鸡同种异体肌腱移植中的免疫排斥反应,增强肌腱愈合强度,减轻肌腱粘连程度。

Objective Triptolide can suppress immunological rejection reaction. To investigate the effect of triptolide on allogenic tendon transplantation in repairing tendon defect in chicken. Methods The defect model of the third toes tendon was established in 64 healthy-cleaning male Leghorn chickens （4-month-old, weighing 1.9-2.3 kg）, which underwent allogenic tendon transplantation for repairing and were divided into 2 groups randomly （n=32）. Triptolide feeding [100 μg/（kg·d）] was given for 3 weeks in the experimental group and normal feeding in the control group. General condition of the chickens was observed after operation. The transplanted tendons were harvested from 4 chickens in each group for gross observation at 1, 2, 3, and 4 weeks after operation; the histological observation was performed at 1 and 3 weeks, and transmission electron microscope observation at 2 and 4 weeks. The blood and tendon were harvested from another 8 chickens in each group for flow cytometry and biomechanical tests respectively at 3 and 6 weeks. Results All chickens survived to the experiment end. Gross observation： with time extending, hyperemia and edema around transplanted tendon were relieved. Rarefaction adhering zone was seen in experimental group, and pyknotic adhering zone in control group. Histological observation： inflammatory reaction in experimental group was slighter than that in control group at 1 and 3 weeks. Transmission electron microscope observation： at 2 and 4 weeks, fibroblasts had big cell nucleus, more euchromatin, and little heterochromatin in experimental group; however, there were small amount of rough endocytoplasmic reticulums with gentle expanded capsular space in control group, which contained sparse content. Flow cytometry test： at 3 and 6 weeks, peripheral blood contained less CD4^＋ and CD8^＋ T lymphocytes in experimental group than in control group, and the ratio of CD4^＋ to CD8^＋ T lymphocyte significantly decreased in experimental group when compared with control group （P 〈 0.05）. Biomechanical examination： at 3 and 6 weeks, the maximum tensile strength in experimental group was bigger than that in control group, and tensile adhesion power in experimental group was smaller than that in control group. There were significant differences in the indexes between 2 groups （P 〈 0.05）. Conclusion Triptolide can suppress immunological rejection reaction, strengthen tendon healing strength, and reduce tendon adhesion in allogenic tendon transplantation.