摘要
目的探讨雷公藤多甙对异体神经移植后靶肌肉萎缩和细胞凋亡的影响。方法20只雄性Wistar大鼠为供体,体重200～250g,取双侧坐骨神经实验。50只雄性SD大鼠为受体,体重200～250g。将受体大鼠制备右侧坐骨神经10mm缺损模型后,随机分为A、B、C、D、E组(n=10):A、B组为新鲜异体神经移植组,C、D组为异体神经雷公藤多甙预处理后再移植组,E组为自体神经移植组。神经移植术后第2天A、E组予生理盐水,B、D组予雷公藤多甙5.0mg/(kg·d),C组予雷公藤多甙2.5mg/(kg·d),时间均为5周。术后3、6周行大体观察、骨骼肌HE染色观察,采用Image-ProPlusv5.2图像处理系统分析肌纤维直径,透射电镜观察骨骼肌超微结构,免疫组织化学检测Bax、Bcl-2表达,TUNEL法检测骨骼肌细胞凋亡。结果术后大鼠全部存活至实验完成。大体观察发现术后3周各组大鼠胫骨前肌均不同程度萎缩;术后6周A组肌肉萎缩严重,其余各组肌肉萎缩呈好转趋势。A组肌湿重、肌纤维直径及Bcl-2表达均显著低于B、C、D、E组(P<0.01),B、C、D组低于E组(P<0.05),B、C、D组间比较差异无统计学意义(P>0.05);A组细胞凋亡指数和Bax表达显著高于B、C、D、E组(P<0.01),B、C、D组高于E组(P<0.05),B、C、D组间差异无统计学意义(P>0.05)。术后3周,光镜下各组肌纤维变细;透射电镜示肌质网不同程度扩张。术后6周,A组光镜示肌纤维间隙增宽、结缔组织明显增生;透射电镜示肌小节结构消失,肌丝溶解严重和残存"Z"线片段;其余组肌原纤维均排列整齐,明带、暗带和肌小节结构清晰。结论异体神经移植术后予雷公藤多甙能减轻靶肌肉萎缩和细胞凋亡,供体神经经雷公藤多甙预处理能减少异体神经移植术后受体免疫抑制剂用量。
Objective To explore the effect of tripterygium glycoside （TG） on the skeletal muscle atrophy and apoptosis after nerve allograft. Methods Twenty Wistar male rats were adopted as donors, weighing 200-250 g, and the sciatic nerves were harvested. Fifty SD male rats were adopted as recipients, weighing 200-250 g. Fifty SD rats were made the models of 10 mm right sciatic nerve defect randomly divided into five groups （n=10）： group A, group B, group C, group D and group E. groups A and B received fresh nerve allograft, groups C and D received sciatic nerve allograft pretreated with TG, and group E received autograft. The SD rats were given medicine for 5 weeks from the second day after the transplantation： groups A and E were given physiological saline, groups B and D TG 5 mg/（kg·d）, and group C TG 2.5 mg/（kg·d）. At 3 and 6 weeks, respectively, after nerve transplantation, general observation was performed; the structure of skeletal muscles was observed by HE staining; the diameter of skeletal muscles was analyzed with Image-Pro Plus v5.2; the ultrastructure of skeletal muscles was observed by TEM; the expressions of Bax and Bcl-2 were detected by immunohistochemical staining; and the apoptosis of skeletal muscles was detected by TUNEL. Results All rats survived to the end of the experiment. In general observation, the skeletal muscles of SD rates atrophied to different degrees 3 weeks after operation. The muscular atrophy in group A was more serious at 6 weeks, and that in the other groups improved. The wet weight, fiber diameter and expression of Bcl-2 in group A were significantly lower than those in groups B, C, D and E （P 〈 0.01）; those in groups B, C and D were lower than those in group E （P 〈 0.05）; and there were no significant differences among groups B, C and D （P 〉 0.05）. The apoptosis index and expression of Bax in group A were significantly higher than those in groups B, C, D and E （P 〈 0.01）; those in groups B, C and D were higher than in group E （P 〈 0.05）; and there were no significant differences among groups B, C and D （P 〉 0.05）. Three weeks after nerve allograft, under the light microscope, the muscle fibers became thin; under the TEM, the sarcoplasmic reticulum was expanded. Six weeks after nerve allograft, Under the light microscope, the gap of the muscle fibers in group A was found to broaden and connective tissue hyperplasia occurred obviously; under the TEM, sarcomere damage, serious silk dissolution and fragmentary Z lines were seen in group A, but the myofibrils were arranged tidily in the other groups, and the light band, dark band and sarcomere were clear. Conclusion TG can decrease the skeletal muscle atrophy and apoptosis after nerve allograft. The donor＇s nerve that is pretreated with TG can reduce the dosage of immunosuppressant for the recipient after allograff.
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2009年第1期101-105,共5页
Chinese Journal of Reparative and Reconstructive Surgery
