山东医药2023，Vol.63Issue(35)：15-18,4.DOI:10.3969/j.issn.1002-266X.2023.35.004
不同复温时点的热打击人骨骼肌细胞损伤情况观察及其机制
Effect and mechanism of heat stress at different rewarming time on damage of human skeletal muscle cells
摘要
Abstract
Objective To explore the the effect and possible mechanism of heat stress at different rewarming time on the damage of human skeletal muscle cells(HSKMC).Methods HSKMC in the logarithmic growth phase were divided into the heat stress group and control group.Cells in the control group were cultured normally,and the heat stress models were established in the heat stress group and then were rewarmed in 37℃and 5%CO2 cell incubator for 0,6,12 and 24 h,respectively.The cell survival rate was detected by CCK-8,the cell ultrastructure was observed by transmission elec-tron microscopy,TRPV4 protein was detected by Western blotting,and TRPV4 mRNA was detected by real-time fluores-cence quantitative PCR.HSKMC in the logarithmic growth phase were randomly divided into the HS group,GSK+HS group,HC+HS group,and NC group.Cells in the NC group were cultured normally,and the heat stress models were es-tablished in the other three groups,and the rewarming time was 12 h.Cells in the GSK+HS group and HC+HS group were given TRPV4 agonist GSK1016790A(500 nmol)and TRPV4 inhibitor HC-067047(5 μmol)at 0.5 h before heat stress,and Ca2+ level was detected by flow cytometry.Results Compared with the control group,the survival rate of cells in heat stress group decreased after rewarming for 0,6,12 and 24 h(P＜0.05).With the extension of rewarming time,the survival rate of cells in the heat stress group decreased successively,and significant difference was found between these two groups(all P＜0.05).Compared with the control group,ultrastructural changes were observed in the cells in the heat stress group after rewarming for 0 and 6 h,which were mainly manifested as cell swelling,enlarged volume,cytoplasmic vacuolation,obvious mitochondrial swelling,degranulation,double ridge loss,widening of perinuclear space,nucleus condensation and contraction,and interruption of some envelope continuity.After rewarming for 12 and 24 h,the cells in the heat stress group even showed nucleolysis and disappearance,nucleus fragmentation,chromatin boundary aggrega-tion,and cytoplasmic structure disintegration and granulation.Compared with the control group,the relative expression levels of TRPV4 mRNA and protein in heat stress group increased after rewarming for 0,6,12 and 24 h(all P＜0.05).The levels of intracellular Ca2+ in the NC group,HC+HS group,HS group and GSK+HS group increased successively,and significant difference was found between groups(all P＜0.05).Conclusions Heat stress can reduce HSKMC survival rate and damage HSKMC structure.The longer rewarming time after heat stress,the greater the damage to HSKMC cells.The mechanism may be related to the promotion of TRPV4 expression and its mediated Ca2+ influx.关键词
瞬时感受器电位香草酸受体亚型4/热打击/复温/钙离子/横纹肌溶解/中暑Key words
TRPV4/heat stress/rewarming/Ca2+/rhabdomyolysis/heatstroke分类
医药卫生引用本文复制引用
刘斌,袁芳芳,豆春丽,肖宝,李霖,李慧,苏磊..不同复温时点的热打击人骨骼肌细胞损伤情况观察及其机制[J].山东医药,2023,63(35):15-18,4.基金项目
湖南省卫健委科研项目重点指导课题项目(202110000190) （202110000190）
睿E(睿意)急诊医学研究专项基金项目(R2020009). （睿意）
