外泌体Homer 1a对氧糖剥夺诱导的PC12细胞损伤的保护作用及机制  

Role of exosome homer-1a in protecting against OGD-induced injury of PC12 cell and its mechanism

作　　者：王远[1] 段发亮[1] 罗明[1] 张严国[1] Wang Yuan;Duan Faliang;Luo Ming;Zhang Yanguo(Department of Neurosurgery,Wuhan First Hospital,Wuhan 430000,Hubei Province,China)

机构地区：[1]武汉市第一医院神经外科,430000

出　　处：《中华老年心脑血管病杂志》2022年第1期87-91,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(81601076);武汉市卫生和计划生育委员会重点项目(WX17B01)。

摘　　要：目的探究外泌体Homer 1a通过Notch信号通路对氧糖剥夺(OGD)诱导的PC12细胞损伤的保护作用及机制。方法选取胰蛋白酶消化传代PC12细胞分为对照组、模型组、空载体组和Homer 1a组(n=9)。对照组正常培养PC12细胞,模型组建立OGD损伤模型,空载体组和Homer 1a组分别将空载体和过表达Homer 1a载体转染到OGD的PC12细胞。采用实时荧光定量PCR检测Homer 1a mRNA表达,MTT法检测细胞活性,流式细胞术检测细胞凋亡率,试剂盒测定乳酸脱氢酶活性、丙二醛水平、超氧化物歧化酶活性,ELISA检测白细胞介素(IL)6、IL-1β、TNF-α水平,Western blot检测Hormer 1a、Notch1、Notch基因细胞内区(NICD)蛋白表达。结果与对照组比较,模型组、空载体组和Homer 1a组Homer 1a表达、细胞凋亡率、乳酸脱氢酶活性、丙二醛、IL-6、IL-1β、TNF-α、Notch1、NICD蛋白表达明显升高,细胞活性、超氧化物歧化酶活性明显降低,差异有统计学意义(P<0.05)。与模型组比较,Homer 1a组Homer 1a mRNA和蛋白表达、细胞活性、超氧化物歧化酶活性明显升高[3.96±0.45 vs 2.18±0.19,3.68±0.72 vs 2.36±0.33,(78.01±8.22)%vs(52.82±7.34)%,(30.80±1.95)U/mg vs(13.67±0.20)U/mg,P<0.05]。Homer 1a组细胞凋亡率、乳酸脱氢酶活性、丙二醛、IL-6、IL-1β、TNF-α、Notch1、NICD蛋白表达明显低于模型组(P<0.05)。结论上调外泌体Homer 1a表达通过抑制Notch信号通路提高OGD诱导的PC12细胞活性,抑制凋亡,同时降低氧化应激和炎症水平。Objective To study the role of exosome Homer-1a in protecting against oxygen glucose deprivation(OGD)-induced injury of PC12 cells by regulating the Notch signaling pathway and its mechanism.Methods Trypsinization passaged 36 PC12 cells were divided into control group,model group,empty vector group and Homer-1a group(9 in each group).The PC12 cells in con-trol group were cultured in normal condition,the OGD-induced PC12 cell injury model was estab-lished for model group,the empty vector and overexpressed Homer-1a vector were transfected in-to OGD-induced PC12 cells of empty vector group and Homer-1a group respectively.The expres-sion of Homer-1a mRNA was detected by RT-qPCR.The viability of PC12 cells was measured with MTT staining.The apoptosis rate of PC12 cells was assayed by flow cytometry.The activity of lactate dehydrogenase and superoxide dismutase and serum level of malondialdehyde were measured with a reagent kit.The expressions of IL-6,IL-1β,TNF-αand Homer-1a,Notch1,Notch gene were detected by ELISA and Western blot respectively.Results The expression levels of Homer-1a,IL-6,IL-1β,TNF-αand Notch1 proteins,opoptosis rate of PC12 cells,activity of lactate dehydrogenase and serum level of malondialdehyde were significantly higher while the activity of PC12 cells and superoxide dismutase was significantly lower in model group,empty vector group and Homer-1a group than in control group(P<0.05).The expression levels of Homer-1a mRNA and protein,activity of PC12 cells and superoxide dismutase were significantly higher in Homer-1a group than in model group(3.96±0.45 vs 2.18±0.19,3.68±0.72 vs 2.36±0.33,78.01%±8.22%vs 52.82%±7.34%,30.80±1.95 U/mg vs 13.67±0.20 U/mg,P<0.05).The opopto-sis rate of PC12 cells,activity of lactate dehydrogenase,serum level of malondialdehyde,and ex-pression levels of IL-6,IL-1β,TNF-αand Notch1 proteins were significantly lower in Homer-1a group than in model group(P<0.05).Conclusion Upregulating the expression of exosome Ho-mer-1a increases the viability of PC12 cells by

关 键 词：外泌体 PC12细胞 L-乳酸脱氢酶 丙二醛 超氧化物歧化酶 白细胞介素6 肿瘤坏死因子α 

分 类 号：R741.02[医药卫生—神经病学与精神病学]