缺氧通过HIF-1α/PCSK9信号途径诱导神经细胞凋亡  

Hypoxia induces neuronal apoptosis through HIF-1α/PCSK9 signaling pathway

作　　者：刘录山[1] 邓懿铭 李清 LIU Lushan;DENG Yiming;LI Qing(Institute of Cardiovascular Disease,University of South China&Key Laboratory for Arteriosclerology of Hunan Province&Human International Scientific and Technological Cooperation Base of Arteriosclerotic Disease,Hengyang,Hunan 421001,China;Department of Pathology,Hunan Polytechnic of Environment and Biology,Hengyang,Hurum 421001,China)

机构地区：[1]南华大学心血管疾病研究所动脉硬化学湖南省重点实验室湖南省动脉硬化性疾病国际科技合作与创新联合实验室,湖南省衡阳市421001 [2]湖南环境生物职业技术学院病理学教研室,湖南省衡阳市421001

出　　处：《中南医学科学杂志》2021年第4期373-378,共6页Medical Science Journal of Central South China

基　　金：国家自然科学基金(81370376);湖南省自然科学基金(2018JJ2343)。

摘　　要：目的研究缺氧是否通过上调缺氧诱导因子1α(HIF-1α)水平而调控前蛋白转化酶枯草溶菌素9(PCSK9)表达并诱导神经细胞凋亡。方法分别用0、125、250、500μmol/L氯化钴(CoCl_(2))处理PC12细胞,蛋白印迹法检测其PCSK9及HIF-1α表达;Hoechst33258核染色及流式细胞术检测CoCl_(2)对PC12细胞凋亡的影响。用250μmol/L CoCl_(2)处理PC12细胞0、6、12、24 h,蛋白印迹法检测其PCSK9及HIF-1α表达。不同浓度HIF-1α激动剂二甲基乙二酰基甘氨酸(DMOG)或HIF-1α抑制剂利非西呱(YC-1)处理PC12细胞24 h,检测PCSK9、HIF-1α、胱天蛋白酶3(Caspase-3)、Caspase-9的表达及对PC12细胞凋亡的影响。PC12细胞经PCSK9 siRNA转染后与250μmol/L CoCl_(2)孵育,检测对PC12细胞凋亡的影响及PCSK9、HIF-1α、Caspase-3、Caspase-9蛋白的表达。结果PCSK9、HIF-1α的表达及细胞凋亡程度呈CoCl_(2)浓度与时间依赖性增高。PC12细胞中HIF-1α和PCSK9的表达呈DMOG浓度依赖性增高,Caspase-3、Caspase-9表达及PC12细胞凋亡率呈DMOG浓度依赖性降低;PC12细胞中HIF-1α和PCSK9的表达呈YC-1浓度依赖性降低,Caspase-3、Caspase-9表达及PC12细胞凋亡率呈YC-1浓度依赖性升高。与空白组及无义RNA组比较,PCSK9 siRNA转染组Caspase-3、Caspase-9表达减少,凋亡程度降低。结论细胞缺氧状态下HIF-1α水平的增高能上调PCSK9表达并调控神经细胞凋亡。Aim To unravel if hypoxia elevates the expression of proprotein convertase subtilisin-kexin type 9(PCSK9)by up-regulating expression of hypoxia-induced factor-1 alpha(HIF-1α)to mediate neuronal apoptosis.Methods PC12 cells were treated with 0,125,250 and 500μmol/L CoCl_(2),respectively,and the expression of PCSK9 and HIF-1αwere detected by western blot.Hoechst33258 nuclear staining and flow cytometry were used to detect the effect of CoCl_(2) on PC12 cell apoptosis.PC12 cells were treated with 250μmol/L CoCl_(2) for 0,6,12 and 24 h,and the expression of PCSK9 and HIF-1αwere detected by western blot.PC12 cells were treated with different concentrations of HIF-1αagonist dimethylglycoglycine(DMOG)or HIF-1αinhibitor rifeciperum(YC-1)for 24 h,and the expression of PCSK9,HIF-1α,Caspase-3 and Caspase-9 and their effects on apoptosis of PC12 cells were detected.PC12 cells were transfected with PCSK9 siRNA and incubated with 250μmol/L CoCl_(2) to detect the effect of PCSK9 apoptosis and the expression of PCSK9,HIF-1α,Caspase-3 and Caspase-9 proteins.Results Expression of PCSK9 and HIF-1αand the degree of apoptosis increased in a time-dependent manner with the concentration of CoCl_(2).Expression of HIF-1αand PCSK9 in PC12 cells increased in a DMOG concentration-dependent manner,while the expression of Caspase-3 and Caspase-9 and the apoptosis rate of PC12 cells decreased in a DMOG concentration-dependent manner.Expression of HIF-1αand PCSK9 in PC12 cells decreased in a YC-1 concentration-dependent manner,while the expression of Caspase-3 and Caspase-9 and the apoptosis rate of PC12 cells increased in a YC-1 concentration-dependent manner.Compared with blank group and meaningless RNA group,the expression of Caspase-3 and Caspase-9 in PCSK9 siRNA transfection group decreased,and the degree of apoptosis decreased.Conclusion Hypoxia increases the level of HIF-1αand promotes the expression of PCSK9 to regulate neuronal apoptosis.

关 键 词：前蛋白转化酶枯草溶菌素9 缺氧诱导因子-1α 凋亡 缺氧 缺氧缺血性脑损伤 

分 类 号：Q255[生物学—细胞生物学]