【摘要】 目的:通过光镜组织学观察评分、电镜超微机构观察和应用逆转录聚合酶链式反应(RT-PCR)技术对内源性胰岛素样生长因子(IGF-Ⅰ、IGF-Ⅱ)mRNA的测定来探讨活血止痛膏剂对骨骼肌钝挫伤修复的作用及其可能机制。方法:将63只兔子随机分为活血止痛膏高剂量组、中剂量组、低剂量组、青鹏膏对照组、外伤对照组,每组12只,另取3只做正常对照组。采用"重物自由落体打击法"制作大白兔骨骼肌急性钝挫伤模型。分别在3天、6天、10天、13天处死大白兔,每组每个时间点处死3只兔子,每只兔子在两条后腿腓肠肌取标本,共计6个标本。结果:(1)光镜组织学观察评分:各个时间点与外伤对照组比较,青鹏膏剂组、活血止痛膏高、中剂量组(P<0.05);青鹏膏和活血止痛膏高剂量组优于活血止痛膏中剂量组(P<0.05)。(2)电镜超微结构观察结果:青鹏膏组、活血止痛膏高、中剂量组较低剂量组和外伤组在3、6天可以更早更多看见激活的肌卫星细胞,在10天和13天青鹏膏组、活血止痛膏高、中剂量组肌组织趋于正常,活血止痛膏低剂量组和外伤组有瘢痕形成。(3)RT-PCR技术检测内源性胰岛素样生长因子(IGF-Ⅰ、IGF-Ⅱ)mRNA的表达量:各个时间点内与外伤对照组比较,活血止痛膏高、中剂量组(P<0.05);活血止痛膏高剂量组优于活血止痛膏中剂量组(P<0.05)。伤后各组IGF-Ⅰ、IGF-ⅡmRNA含量在3、6、10、13天升高,其中第6天达高峰值。结论:(1)活血止痛膏可有效的改善兔骨骼肌钝挫伤后的组织学愈合过程和愈合质量。(2)活血止痛膏能促进骨骼肌钝挫伤后卫星细胞的激活,进一步促进新肌生成。(3)内源性IGF-I,IGF-II的mRNA随着兔骨骼肌钝挫伤不同修复时期,表达量也相应变化。(4)活血止痛膏对兔骨骼肌钝挫伤后的内源性IGF-I,IGF-II的mRNA表达量上调有明显的促进作用,从而有利于骨骼肌挫伤后愈合;并且该表达量与活血止痛膏有效浓度呈正相关。更多还原

【Abstract】 Objective:To investigate the role and possible mechanism of Huoxue Zhitong Plaster by light microscope,electron microscope and the measurement of endogenous insulin-like growth factor(IGF-Ⅰ,IGF-Ⅱ) mRNA using Reverse Transcription Polymerase Chain Reaction(RT-PCR)technique.Methods:63 rabbits were randomly divided into Huoxue Zhitong Plaster high dose group,middle dose group,low dose group,Qingpeng Plaster group,trauma control group.Each group contained 12 rabbits,and there were 3 rabbits in normal control.The method of Free-falling weights was used to make the model with acute skeletal muscle contusion.The rabbits were killed on the 3rd,6th,10th and 13th day respectively,3 rabbits were killed at each time point in each group,specimens were taken on the gastrocnemius of the legs.Results:(1)Light microscope histological score:compared with trauma control group,Qingpeng Plaster group,Huoxue Zhitong Plaster high dose group and middle dose group had significant difference(P<0.05);Qingpeng Plaster group and Huoxue Zhitong Plaster high dose group were better than Huoxue Zhitong Plaster middle dose group(P<0.05).(2)Ultrastructural electron microscope observations: compared with low dose group and trauma control group,more activated muscle satellite cells can be found on Qingpeng Plaster group,Huoxue Zhitong Plaster high dose group and middle dose group in 3rd or 6th day.Muscle tissues were nomal in Qingpeng Plaster group,Huoxue Zhitong Plaster high dose group and middle dose group and scar formation can be seen in Huoxue Zhitong Plaster low dose group and trauma control group.(3)RT-PCR detection of endogenous insulin-like growth factor(IGF-Ⅰ,IGF-Ⅱ) mRNA expression:at each time point compared with trauma control group,Huoxue Zhitong Plaster high dose group and middle dose group had significant difference(P<0.05);Huoxue Zhitong Plaster high dose group was better than Huoxue Zhitong Plaster middle dose group(P<0.05).IGF-1 and IGF-2 levels increased on 3rd,6th,10th,13th day in each group,and reached a peak value on the 6th day.Conclusion:(1)Huoxue Zhitong Plaster can effectively improve the rabbit skeletal muscle tissue after contusion healing process and healing quality.(2)Huoxue Zhitong Plaster can promote the activation of skeletal muscle satellite cells after injury,and further promote the formation of new muscle.(3)the expression of mRNA by endogenous IGF-I and IGF-II changes are according with different recovery periods.(4)Huoxue Zhitong Plaster can significantly promote the expression of mRNA by endogenous IGF-I of rabbit skeletal muscle contusion,thus contributing to skeletal muscle contusion healing;and the expression amount was positively correlated with effective concentration of Huoxue Zhitong Plaster.更多还原