丹曲林对严重烧伤大鼠骨骼肌损害的治疗作用及机制  

Effect and mechanism of dantrolene on skeletal muscle of rats with severe scald injury

作　　者：马丽[1] 柴家科[1] 褚万立[1] 段红杰[1] 李大伟[1] 郁永辉[1] 

机构地区：[1]解放军总医院第一附属医院烧伤整形科,北京100048

出　　处：《中华医学杂志》2014年第14期1087-1091,共5页National Medical Journal of China

基　　金：国家自然科学基金（81071544、81120108014）

摘　　要：目的：探讨丹曲林对严重烧伤大鼠骨骼肌损害的治疗作用及可能机制。方法选取清洁级Wistar雄性大鼠56只，按照随机数字表法分为对照组8只、致伤组及丹曲林组各24只（两组3个时相点各8只）。致伤组和丹曲林组大鼠在麻醉后与其背部及腹部行50％总体表面积（ TBSA ）Ⅲ度烫伤，伤后腹腔注射林格平衡液抗休克，致伤组尾静脉注射5％甘露醇，丹曲林组尾静脉注射丹曲林钠2 mg／kg（溶于5％甘露醇）。对照组行假烫伤，其余操作同致伤组。于伤后第1、4、7天收集各组大鼠胫骨前肌组织，透射电镜观察组织超微结构，电子探针X射线显微分析法检测肌细胞微区Ca2＋含量，蛋白印迹方法检测胫骨前肌总蛋白中钙蛋白酶1、2表达，酶法检测胫骨前肌中钙蛋白酶的活性。结果致伤组胫骨前肌肌丝伤后第1天出现排列混杂，第7天出现局部Z线消失。而丹曲林组第1、4天肌组织超微结构与对照相比无显著变化，仅在第7天肌丝存现排列卷曲、轻度断裂。伤后第1、4天致伤组胫骨前肌细胞质Ca2＋水平显著高于对照组[（0.964±0.060）、（0.639±0.067）比（0.266±0.029）μmol／L]，肌浆网Ca2＋水平显著低于对照组[（0.368±0.060）、（0.814±0.089）比（1.337±0.112）μmol／L]（均P＜0.05）；而丹曲林组细胞质和肌浆网Ca2＋水平[（0.310±0.069）、（0.490±0.039）和（1.241±0.073）、（1.161±0.094）μmol／L]与对照组差异均无统计学意义（均P＞0.05）。伤后第1、4天致伤组胫骨前肌钙蛋白酶1和钙蛋白酶2相对表达量均显著高于对照组（1.371±0.034、1.214±0.030比0.838±0.017和1.464±0.015、1.390±0.023比0.806±0.026）（均P＜0.05），而丹曲林组钙蛋白酶1和钙蛋白酶2相对表达量（0.984±0.031、0.935±0.023和0.836±0.014与0.741±0.020）均显著低于致伤组（均P＜0.05）。伤后第1、4、7天致伤组和丹曲林组钙蛋白酶活性分�Objective To explore the effect and mechanism of ryanodine receptor antagonist dantrolene on skeletal muscle of rats with severe scald injury.Methods A total of 56 Wistar rats were divided into control , scald and dantrolene treatment groups according to a random digital table.Rats in scald and dantrolene treatment groups were subject to 50%total body surface area （ TBSA） full-thickness scald by a 12-second immersion of back and a 6-second immersion of abdomen in 94 ℃water and then received an intraperitoneal injection of Ringer′s solution.At the same time , the rats in scald group received 5%mannitol through caudal vein while those in dantrolene treatment group received dantrolene 2 mg/kg （ dissolved in 5%mannitol ）.Rats in control group were sham-injured through an immersion of back and abdomen into 37 ℃warm water.Tibialis anterior muscle samples were harvested at Days 1, 4 and 7 post-scalding.Changes of skeletal muscle ultrastructure were observed by transmission electron microscope , subcellular calcium ion （ Ca2＋） contents of skeletal muscle （ including cytoplasm , mitochondria ＆amp; sarcoplasm reticulum ） were detected by electron probe X-ray microanalysis （ EPMA） and the levels of calpain-1 and calpain-2 protein were determined by Western blot.And the activities of calpain were detected by enzyme-linked immunosorbent assay.Results In scald group , assorted arrangement appeared immediately at Day 1 post-injury and partial disappearance of Z lines at Day 7 post-injury.There were no significant ultrastructure changes in dantrolene treatment group at Day 1 and 4 post-injury.Curled filament and mild fracture occurred merely in dantrolene treatment group at Day 7 post-injury.The cytoplasmic contents of Ca 2＋ were significantly higher in scald group than those in control group at Day 1 and 4 （（0.964 ±0.060）, （0.639 ± 0.067） vs （0.266 ±0.029） μmol/L respectively, all P〈0.05） while the contents of Ca2＋within sarcoplasm reticulum were obviously lower in scald g

关 键 词：丹曲林 烧伤 肌 骨骼 大鼠 钙蛋白酶 

分 类 号：R644[医药卫生—外科学]