模拟微重力对HaCaT细胞创伤修复相关蛋白和基因表达的影响  

Effects of Simulated Microgravity on Expressions of Wound Healing Related Proteins and Genes in HaCaT Cells

作　　者：林晶晶 孙培鸣[1] 张涛[1] 徐冰心[2] 段育忠[1] 周金莲[3] 杨鹤鸣[1] 李成林[1] 崔彦[1] LIN Jing-jing;SUN Pei-ming;ZHANG Tao;XU Bing-xin;DUAN Yu-zhong;ZHOU Jin-lian;YANG He-ming;LI Cheng-lin;CUI Yan(Department of General Surgery,306 Hospital of PLA,Beijing 100101,China;Special Medical Laboratory Center,306 Hospital of PLA,Beijing 100101,China;Department of Pathology,306 Hospital of PLA,Beijing 100101,China)

机构地区：[1]解放军第306医院普通外科,北京100101 [2]解放军第306医院特种医学实验研究中心,北京100101 [3]解放军第306医院病理科,北京100101

出　　处：《解放军医药杂志》2020年第1期1-5,共5页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：全军试验技术研究计划重点项目(SYFD1500128)

摘　　要：目的研究模拟微重力环境对人角质细胞(HaCaT细胞)创伤修复相关蛋白和基因表达的影响。方法应用RCCS系统进行模拟微重力细胞培养实验,将HaCaT细胞随机分为模拟微重力(SMG)组和正常重力对照(NG)组,培养32、36和42 h后分别收集细胞,检测热休克蛋白70(HSP70)、基质金属蛋白酶-9(MMP-9)和Caspase-3的mRNA及蛋白表达水平变化,透射电镜观察2组培养42 h后细胞超微结构变化。结果SMG组HaCaT细胞培养32、36 h HSP70 mRNA表达量,培养32、36和42 h MMP-9 mRNA及培养36 h Caspase-3 mRNA表达水平均高于NG组(P<0.05)。蛋白免疫印迹法检测结果显示,HaCaT细胞经微重力培养32、36和42 h后HSP70和MMP-9蛋白表达趋势与相应蛋白mRNA表达一致,而Caspase-3蛋白表达趋势与相应mRNA表达存在一定差异。透射电镜观察发现,模拟微重力环境下培养42 h后的HaCaT细胞出现皱缩表现,线粒体嵴出现断裂,结构模糊,部分线粒体出现空泡。结论模拟微重力环境下HaCaT细胞HSP70、MMP-9及Caspase-3表达水平上调,细胞超微结构发生明显病理性改变。Objective To investigate effects of simulated microgravity on expressions of wound healing related proteins and genes in human keratinocyte HaCaT cells.Methods The rotary cell culture system(RCCS)was used to simulate microgravity environment.Human keratinocyte HaCaT cells were randomly divided into simulated microgravity group(SMG)and normal gravity group(NG),and cells were collected after culture for 32,36 and 42 h,and then the mRNA and protein expression levels of HSP70,MMP-9 and Caspase-3 were detected,and ultrastructure changes of HaCaT cells after culture for 42 h were observed by transmission electron microscopy(TEM)in two groups.Results HSP70 mRNA expressions after culture for 32 and 36 h,MMP-9 mRNA expressions after culture for 32,36 and 42 h and Caspase-3 mRNA expression after culture for 36 h of HaCaT cells in SMG group were significantly higher than those in NG group(P<0.05).Western blotting results revealed that tendency of HSP70 and MMP-9 protein expressions were consistent with those of corresponding protein expressions,while tendency of Caspase-3 protein expressions were significant differences compared with those of corresponding mRNA expressions after 32,36 and 42 h of culture under simulated microgravity.TEM observation showed that HaCaT cells after 42 h culture under simulated microgravity were shrank,and mitochondrial cristae was broken with blurred structure and vacuoles in some mitochondria.Conclusion RCCS simulated microgravity environment may enhance expressions of HSP70,MMP-9 and Caspase-3 of HaCaT cells and induce obviously pathological changes in cell ultrastructure.

关 键 词：失重模拟 HACAT细胞 热休克蛋白70 基质金属蛋白酶-9 Caspase-3 超微结构 

分 类 号：R851[医药卫生—航空、航天与航海医学]