Huang Yingru, Jiang Dianming, Chen Lu, Jiang Hua
Department of Orthopaedics, Traditional Chinese Medical College of Chongqing Medical University, Chongqing, 401331, P.R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2009 Jan;23(1):101-5.
To explore the effect of tripterygium glycoside (TG) on the skeletal muscle atrophy and apoptosis after nerve allograft.
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 x d), and group C TG 2.5 mg/(kg x 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.
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.
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 allograft.
探讨雷公藤多苷(TG)对异体神经移植后骨骼肌萎缩及凋亡的影响。
选取20只体重200 - 250 g的Wistar雄性大鼠作为供体,切取坐骨神经。选取50只体重200 - 250 g的SD雄性大鼠作为受体。将50只SD大鼠制作成10 mm右侧坐骨神经缺损模型,随机分为五组(n = 10):A组、B组、C组、D组和E组。A组和B组接受新鲜神经异体移植,C组和D组接受经TG预处理的坐骨神经异体移植,E组接受自体移植。SD大鼠于移植后第二天开始给药5周:A组和E组给予生理盐水,B组和D组给予TG 5 mg/(kg·d),C组给予TG 2.5 mg/(kg·d)。分别于神经移植后3周和6周进行一般观察;采用苏木精-伊红(HE)染色观察骨骼肌结构;用Image-Pro Plus v5.2分析骨骼肌直径;通过透射电子显微镜(TEM)观察骨骼肌超微结构;采用免疫组织化学染色检测Bax和Bcl-2的表达;用TUNEL法检测骨骼肌凋亡情况。
所有大鼠均存活至实验结束。一般观察发现,术后3周SD大鼠骨骼肌均有不同程度萎缩。6周时A组肌肉萎缩更严重,其他组有所改善。A组的湿重、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组之间差异无统计学意义(P > 0.05)。A组的凋亡指数及Bax表达均显著高于B、C、D、E组(P < 0.01);B、C、D组高于E组(P < 0.05);B、C、D组之间差异无统计学意义(P > 0.05)。异体神经移植后3周,光镜下可见肌纤维变细;电镜下可见肌浆网扩张。异体神经移植后6周,光镜下发现A组肌纤维间隙增宽,结缔组织增生明显;电镜下可见A组肌节损伤、肌丝严重溶解及Z线断裂,而其他组肌原纤维排列整齐,明带、暗带及肌节清晰。
TG可减轻异体神经移植后骨骼肌萎缩及凋亡。经TG预处理的供体神经可降低异体移植后受体的免疫抑制剂用量。