远隔缺血预适应血清外泌体增加SH-SY5Y细胞糖氧剥夺耐受的生物信息学分析  

Bioinformatics analysis of RIPC serum exosomes to increase oxygen-glucose deprivation tolerance in SH-SY5Y cells

作　　者：邹明新[1] 刘晓蕾[2] 刘娜 崔钧贺 张璞 邵国[3] 时静华[2] Zou Mingxin;Liu Xiaolei;Liu Na;Cui Junhe;Zhang Pu;Shao Guo;Shi Jinghua(School of Biological Science and Technology,Baotou Normal College,Inner Mongolia University of Science and Technology,Baotou 014030,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古科技大学包头师范学院生物科学与技术学院,014030 [2]包头医学院内蒙古自治区低氧转化医学重点实验室 [3]首都医科大学宣武医院低氧适应转化医学北京重点实验室

出　　处：《中华老年心脑血管病杂志》2021年第3期306-309,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(81660307,82060337);内蒙古自治区自然科学基金(2018LH03029);内蒙古自治区高等学校科学研究项目(NJZY18384)。

摘　　要：目的研究远隔缺血预适应(RIPC)血清外泌体对SH-SY5Y细胞基因表达的影响,探讨差异表达基因对神经细胞在糖氧剥夺(OGD)耐受中的作用。方法RIPC前后分离人血清外泌体,取RIPC前外泌体(HuE-C)和RIPC后外泌体(HuE-RIPC)。将SH-SY5Y细胞分为空白组(正常培养细胞),对照组(培养液加入HuE-C)和RIPC组(培养液加入HuE-RIPC),采用高通量测序各组细胞基因表达并进行生物信息学分析,将3组细胞置于OGD条件下,检测细胞活力,采用实时定量PCR及Western blot检测与神经低氧/缺血相关的基因表达。结果生物信息学显示,RIPC组热休克蛋白(HSP)基因差异表达增加。MTS检测发现,OGD条件下,RIPC组细胞活力明显高于空白组(0.673±0.056 vs 0.481±0.027,P<0.01)。实时定量PCR和Western blot检测显示,OGD条件下,RIPC组HSP70 mRNA表达和蛋白表达明显高于空白组和对照组(P<0.05)。结论人RIPC血清外泌体可引起众多基因表达变化,其中HSP70表达增加可能是其低氧耐受增加的一个原因。Objective To study the effect of RIPC serum exosomes on gene expression in SH-SY5Y cells and the role of differentially expressed genes in OGD tolerance of SH-SY5Y cells.Methods HuE-C and HuE-RIPC were obtained.The SH-SY5Y cells were divided into blank group,control group and RIPC group.The cells in blank group were cultured in normal medium,those in control group were cultured in culture fluid+HuE-C and those in RIPC group were cultured in culture fluid+HuE-RIPC.The gene expressions in different groups were detected by high throughput sequencing and analyzed by bioinformatics analysis.The vitality of SH-SY5Y cells under OGD was measured by MTS.The hypoxia/ischemia-related gene expressions were detected by RT-PCR and Western blot respectively.Results Bioinformatics analysis showed that the expression of HSP gene was increased in RIPC group.MTS displayed that the vitality of SH-SY5Y cells under OGD was significantly higher in RIPC group than in blank group(0.673±0.056 vs 0.481±0.027,P<0.01).RT-PCR and Western blot demonstrated that the expression level of HSP70 mRNA and protein was significantly higher in RIPC group than in blank group(P<0.05).Conclusion RIPC serum exosomes can lead to changes in expression of many genes and elevated expression of HSP70 is one of the reasons why the hypoxia tolerance in RIPC serum exosomes is increased.

关 键 词：缺血预处理 外泌体 基因表达 HSP70热休克蛋白质类 

分 类 号：Q811.4[生物学—生物工程] R743.3[医药卫生—神经病学与精神病学]