Circ_HECTD1调节miR-135a-5p/TP53INP1轴对OGD/R诱导的海马神经元损伤的影响  

Influence of Circ_HECTD1on OGD/R-induced hippocampal neuronal damage by regulating the miR-135a-5p/TP53INP1 axis

作　　者：逯琴[1] 张文敏[1] 赵敏[1] LU Qin;ZHANG Wenmin;ZHAO Min(Department of Neurology,Linfen People’s Hospital,Linfen 041000,China)

机构地区：[1]临汾市人民医院神经内科,山西临汾041000

出　　处：《中风与神经疾病杂志》2022年第6期503-509,共7页Journal of Apoplexy and Nervous Diseases

摘　　要：目的通过进行体外氧葡萄糖剥夺/复氧(OGD/R)诱导海马神经元损伤,观察环状RNA含HECT结构域的E3泛素连接酶1(Circ_HECTD1)对HT22细胞增殖、凋亡的影响以及对微小RNA-135a-5p(miR-135a-5p)/肿瘤蛋白53诱导型核蛋白1(TP53INP1)轴的调控机制。方法常规培养HT22细胞,将细胞分为Con组、OGD/R组、si-NC组、si-Circ_HECTD1组、miR-NC组、miR-135a-5p组、si-Circ_HECTD1+anti-miR-NC组、si-Circ_HECTD1+anti-miR-135a-5p组。qRT-PCR法检测Circ_HECTD1、miR-135a-5p、TP53INP1 mRNA表达;MTT检测细胞活性;流式细胞术检测细胞凋亡;双荧光素酶报告基因实验验证Circ_HECTD1与miR-135a-5p、miR-135a-5p与TP53INP1的靶向关系;Western blot法检测Bax、Bcl-2、TP53INP1蛋白表达。结果OGD/R诱导后HT22细胞中Circ_HECTD1与TP53INP1表达上调,miR-135a-5p表达下调,细胞存活率和Bcl-2蛋白表达显著下降,凋亡率和Bax蛋白表达显著升高(均P<0.05)。沉默Circ_HECTD1表达能够显著上调OGD/R诱导的HT22细胞中miR-135a-5p表达,下调TP53INP1表达,提高细胞存活率和Bcl-2蛋白表达,降低细胞凋亡率和Bax蛋白表达(均P<0.05)。Circ_HECTD1与miR-135a-5p、miR-135a-5p与TP53INP1之间均存在靶向关系。过表达miR-135a-5p能够显著下调TP53INP1表达,提高细胞存活率、Bcl-2蛋白表达,降低细胞凋亡率、Bax蛋白表达(均P<0.05)。而抑制miR-135a-5p表达能够部分逆转沉默Circ_HECTD1对HT22细胞损伤的保护作用。结论沉默Circ_HECTD1可通过调节miR-135a-5p/TP53INP1轴,促进细胞存活,抑制细胞凋亡,保护OGD/R诱导的海马神经元损伤。Objective To observe the influences of circular RNA HECT domain E3 ubiquitin ligase 1(Circ_HECTD1)on the proliferation and apoptosis of HT22 cells and its regulatory mechanism on the microRNA-135 a-5 p(miR-135a-5p)/tumor protein 53-induced nuclear protein 1(TP53 INP1)axis by in vitro oxygen-glucose deprivation/reoxygenation(OGD/R)-induced hippocampal neuron damage.Methods HT22 cells were routinely cultured,and the cells were separated into Con group,OGD/R group,si-NC group,and si-Circ_HECTD1 group,miR-NC group,miR-135a-5p group,si-Circ_HECTD1+anti-miR-NC group,and si-Circ_HECTD1+anti-miR-135a-5p group.qRT-PCR method was used to detect the expression of Circ_HECTD1,miR-135a-5p and TP53 INP1 mRNA;MTT was used to detect cell viability;flow cytometry was used to detect apoptosis;dual-luciferase reporter gene experiment was applied to verify the targeting relationship between Circ_HECTD1 and miR-135a-5p,miR-135a-5p and TP53 INP1;Western blot was applied to measure the protein expressions of Bax,Bcl-2 and TP53 INP1.Results After OGD/R induction,the expressions of Circ_HECTD1 and TP53 INP1 in HT22 cells were up-regulated,the expression of miR-135a-5p was down-regulated,the cell survival rate and the expression of Bcl-2 protein were remarkably decreased,the apoptosis rate and the expression of Bax protein were remarkably increased(all P<0.05).Silencing the expression of Circ_HECTD1 could remarkably up-regulate the expression of miR-135a-5p in OGD/R-induced HT22 cells,down-regulate the expression of TP53 INP1,increase cell survival rate and Bax protein expression,decrease cell apoptosis rate and Bcl-2 protein expression(all P<0.05).There was a targeting relationship between Circ_HECTD1 and miR-135a-5p,between miR-135a-5p and TP53 INP1.Overexpression of miR-135a-5p could remarkably down-regulate the expression of TP53 INP1,increase cell survival rate and Bcl-2 protein expression,decrease cell apoptosis rate and Bax protein expression(all P<0.05).Inhibition of miR-135a-5p expression could partially reverse the protective ef

关 键 词：环状RNA含HECT结构域的E3泛素连接酶1 微小RNA-135a-5p 肿瘤蛋白53诱导型核蛋白1 氧葡萄糖剥夺/复氧 海马神经元 

分 类 号：R285.5[医药卫生—中药学]