摘要
目的:建立大鼠胫前肌深度冻伤模型了解其病理变化特点,探索组织工程化成肌细胞移植的适宜动物模型。方法:实验于2003-09/2004-10在四川大学华西医院修复重建实验室完成。选择5月龄SD大鼠30只,随机分10组,正常对照组、术后1,3,5,7,10d,2,4,8,12周组,每组3只,6个样本。采用液氮冷冻后的金属条按一定时间放置于大鼠胫前肌表面,造成胫前肌深度冻伤,测定大鼠胫前肌冻伤前后肌重、磷酸肌酸激酶及同工酶的变化。采用苏木精-伊红染色观察骨骼肌受冻伤后肌组织病理变化;采用免疫组织化学Ⅳ型胶原染色观察受冻肌组织基膜损伤及再生情况;采用Mallory三色染色法观察冻伤组织修复时胶原增生情况,以了解其纤维化愈合的程度。结果:纳入动物30只,均进入结果分析。①肌重变化:冻伤前,冻伤后1,3,5,7,10d,2,4,8,12周肌重平均值分别为0.5889,0.7066,0.7302,0.6735,0.5929,0.5687,0.5496,0.4099,0.3493,0.3036g。术后1,3,5d组肌重均增高(P<0.05);术后7,10d组无显著差异(P>0.05);而术后2,4,8,12周组均降低(P<0.05)。即冻伤后的胫前肌经历最初5d肌肉肿胀,肌重增加,2周开始肌重呈明显下降的过程。②磷酸肌酸激酶及磷酸肌酸激酶-MM含量:术后磷酸肌酸激酶及磷酸肌酸激酶-MM较冻伤前明显下降,术后3d呈逐渐降低趋势。③组织学观察:大鼠胫前肌经深度冻伤后,肌组织变性、坏死,肌纤维的基膜保持较完整,深度冻伤局部缺乏成肌细胞,修复以脂肪性变和纤维增生为主,后期肌肉明显萎缩。结论:大鼠胫前肌深度冻伤模型适合作为成肌细胞移植的动物模型。
AIM： To establish the models of severely cryodamaged tibialis anterior muscles in rats, study the characters of pathological changes and investigate the suitable animal models of myoblast transplantation.
METHODS： The experiment was carried out at the Repair and Reconstrution Laboratory of Huaxi Hospital, Siehuan University from September 2003 to October 2004. Thirty SD rats of five-month old were divided into 10 groups randomly （1, 3, 5, 7, 10 days, 2, 4, 8, 12 weeks after treating and control group）, with three rats and six samples in each group. The tibialis anterior muscles of rats were severely cryodamaged by placing the metal strip （freezed in liquid nitrogen previously） on the surface. The changes of tibialis anterior muscles＇weights, creatine phosphokinase （CK） and creatine kinase-MM isoenzyme （CK-MM） were detected before and after cryodamage respectively. The pathologic changes of skeletal muscle tissues were observed with the hematoxylin-eosin staining; The injury and regeneration status of cryodamaged muscle tissues were observed by the immunohistochemistry Tape-Ⅳ collagen staining; The Mallory trichrome staining was used to observe the proliferation status of collagen when the cryodamaged tissues repaired so as to comprehend the healing degree of fibrosis.
RESULTS： Thirty rats were entered into the result analysis.①The changes of muscle weights： Before eryodamage and at 1, 3, 5, 7, 10 days, 2, 4, 8, 12 weeks after cryodamage, the average values were 0.588 9 g, 0.706 6 g, 0.730 2 g, 0.673 5 g, 0.592 9 g, 0,568 7 g, 0.549 6 g, 0.409 9 g, 0.349 3 g, 0.303 6 g. The muscle weights increased at 1,3,5 days after operation （P 〈 0.05）, and decreased at 2,4,8,12 weeks after operation （P 〈 0.05）. There was no significant difference between 7 days and 10 days after operation （P 〉 0.05）. The eryodamaged tibialis anterior muscles swelled at the initial 5 days with the increased weights, and the weights remarkably decreased from the second weeks. ②The values of CK and CK-MM： Compared with before cryodamage, the values of CK and CK-MM descended obviously after operation, and showed the gradual decrease tendency at 3 days after operation. ③Histological observation： After the severe cryodsmage, the tibialis anterior muscles underwent degeneration and necrosis. The basement membranes of muscle fibers were kept intact. With the deficiency of myoblast in local severely cryodamaged area, fatty and fibrinoid degeneration were the major modes in the repair process. The muscle atrophy was clearly showed at the later period.
CONCLUSION： Severely cryodamaged tibialis anterior muscle in rats can be used for the animal models of transplanted myoblast.
出处
《中国临床康复》
CAS
CSCD
北大核心
2006年第16期82-85,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家高技术研究发展计划(八六三计划)(2001AA216011)~~
