热打击诱导骨骼肌细胞胀亡而非凋亡  

Heat stroke induces oncosis rather than apoptosis in skeletal muscle cells

作　　者：豆春丽 袁芳芳 刘斌 李慧 苏磊 Dou Chun-Li;Yuan Fang-Fang;Liu Bin;Li Hui;Su Lei(Graduate School,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Department of Hematology,the Third Xiangya Hospital of Central South University,Changsha 410013,China;Department of Emergency,Changsha First Hospital,Changsha 410005,China;Department of Intensive Care Unit,Guangzhou General Hospital of Southern Theater Command,Guangzhou 510010,China;Key Laboratory of Tropical Zone Trauma Care and Tissue Repair of PLA,Guangzhou 510010,China)

机构地区：[1]广州中医药大学研究生院,广州510006 [2]中南大学湘雅三医院血液科,长沙410013 [3]长沙市第一医院急诊科,长沙410005 [4]南部战区总医院重症医学科广州,510010 [5]全军热区创伤救治与组织修复重点实验室,广州510010

出　　处：《解放军医学杂志》2020年第10期1047-1051,共5页Medical Journal of Chinese People's Liberation Army

基　　金：军队后勤科研计划项目(18CXZ030,CWH17L020,17CXZ008)。

摘　　要：目的探讨热应激过程中骨骼肌细胞发生损伤的具体形式,以及重症中暑合并横纹肌溶解的机制。方法将人骨骼肌细胞(HSKMC)分为对照组与热打击组,对照组细胞于37℃、5%CO2细胞培养箱中孵育;热打击组细胞于43℃、5%CO2细胞培养箱中孵育2 h构建热打击损伤模型,然后置于37℃、5%CO2细胞培养箱中孵育。待孵育0、6、12、24 h后,采用CCK-8法检测细胞活力,LDH法检测细胞毒性,透射电子显微镜观察细胞超微结构的变化情况,AnnexinⅤ-FITC/PI双染色法检测双阳性细胞率,Western blotting检测细胞中porimin、caspase-3蛋白的表达情况。结果与对照组比较,热打击后复温0h HSKMC细胞活力即开始下降,细胞毒性即增加,且均呈时间依赖性(P<0.05)。对照组细胞表面具有丰富的微绒毛,线粒体、内质网等细胞器结构清晰,核大浓染,形态不规则,染色质浓密,核仁大而清晰;热打击组细胞肿胀、体积变大,胞质空泡化,线粒体明显肿胀,脱颗粒,双嵴消失,核周间隙增宽,细胞核凝聚、固缩,部分包膜连续性中断,且后期伴随有核仁溶解消失,细胞核碎裂、染色质边集,以及胞质结构崩解呈颗粒状等超微结构改变,符合细胞胀亡样表现。与对照组比较,热打击组porimin蛋白的表达水平及双阳性细胞率明显增高(P<0.05),caspase-3蛋白的表达水平差异无统计学意义(P>0.05)。结论热打击可诱导骨骼肌细胞胀亡而非凋亡。Objective To investigate the oncosis of skeletal muscle cells during heatstroke,and the mechanism severe heatstroke associated with rhabdomyolysis.Methods The human skeletal muscle cells(HSKMC)were divided into control group and heat shock group.The control group was incubated at 37℃,5%CO2 cell incubator,while the heat shock group was exposed at 43℃,5%CO2 cell incubator for 2 hours,followed by incubating at 37℃,5%CO2 cell incubator.At incubation point of 0,6,12,and 24 hours,the CCK-8 method was used to detect cell viability;the LDH method was used to detect cytotoxicity;the transmission electron microscope was used to detect cell ultrastructure changes;AnnexinⅤ-FITC/PI double staining method was used to detect double-positive cell rate;Western blotting method was used to detect porimin and caspase-3 protein expression.Results Compared with the control group,HSKMC cell viability decreased with cytotoxicity increased at 0 h after rewarming in a timedependent manner(P<0.05).The ultrastructure of cells in the control group has abundant microvilli on the surface;mitochondria,endoplasmic reticulum,and other organelles have clear structures;densely stained nuclei,irregular shapes,dense chromatin,and large and clear nucleoli are observed.However,cells in the heat shock group are swollen and bulky;the cytoplasm is vacuolated with swollen degranulated mitochondria that lost the double cristae;the perinuclear space widens;the nucleus condenses and shrinks;the continuity of some envelopes is interrupted.In the later stage,the nucleolus is dissolved and disappeared,the nucleus is fragmented,chromatin is marginalized,and the cytoplasmic structure is disintegrated,showing granular ultrastructural changes,which are consistent with cell swelling and death.Compared with the control group,the porimin protein expression and double-positive cell rate in the heat shock group were significantly increased(P<0.05),but there was no significant difference in the caspase-3 protein(P>0.05).Conclusion Heat stroke-induced oncosis rathe

关 键 词：中暑 骨骼肌细胞 细胞胀亡 

分 类 号：R594.12[医药卫生—内科学]