高糖通过抑制Bmal1调控的自噬加重细胞缺氧复氧损伤  

High glucose aggravates hypoxia reoxygenation injury by inhibiting autophagy regulated by Bmal1

作　　者：高文蔚[1] 袁泉[2] 江梦[2] 刘恋[2] 赵博[2] GAO Wenwei;YUAN Quan;JIANG Meng;LIU Lian;ZHAO Bo(Department of Critical Care Medicine,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Anesthesiology,Renmin Hospital of Wuhan University)

机构地区：[1]武汉大学人民医院重症医学科,武汉430060 [2]武汉大学人民医院麻醉科

出　　处：《山西医科大学学报》2020年第1期65-68,共4页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81901994);湖北省自然科学基金资助项目(2019CFC847)。

摘　　要：目的 探讨高糖诱导氧化应激后,Bmal1调控的自噬在细胞缺氧复氧损伤中的作用。方法 采用随机数字表法将PC-12细胞分为4组(n=6):低糖组(NG组)、低糖缺氧复氧组(NH/R组)、高糖组(HG组)和高糖缺氧复氧组(HH/R组)。采用高糖刺激24 h建立高糖模型,缺氧3 h复氧6 h建立缺氧复氧损伤模型。LDH检测细胞活性;ROS,SOD和MDA检测氧化应激水平;Annexin V-FITC/PI流式细胞术检测细胞凋亡率;Western blot检测Bmall,Beclin-1的表达;电镜观察自噬小体数量。结果 与NG组相比,NH/R组SOD、Bmall降低,LDH、ROS、MDA、凋亡率、Beclin-1升高,自噬小体增多(P<0.05);与NCG组相比,HCG组SOD、Bmall、Beclin-1降低,自噬小体减少,LDH、ROS、MDA、凋亡率升高(P<0.05);与NH/R组相比,HH/R组SOD、Bmall、Beclin-1降低,自噬小体减少,LDH、ROS、MDA、凋亡率升高(P<0.05);与IHG组相比,HH/R组SOD、Bmall降低,LDH、ROS、MDA、凋亡率升高(P<0.05),Beclin-1、自噬小体无统计学差异(P>0.05)。结论 高糖诱导的氧化应激抑制时钟基因Bmal1的表达,进而导致自噬功能降低,缺氧复氧损伤加重。Objective To investigate the role of oxidative stress induced by high glucose on autophagy regulated by Bmal1 in PC-12 cells under hypoxia reoxygenation injury.Methods PC-12 cells were randomly divided into four groups(n=6):normal group(NG),hypoxia reoxygenation group(NH/R),high glucose group(HG),high glucose+hypoxia reoxygenation group(HH/R).The high glucose model was established by 25 mmol/L glucose for 24 h and the hypoxia reoxygenation model was established by hypoxia for 3 h and reoxygenation for 6 h.Lactate dehydrogenase(LDH)was used to evaluate cell viability,the enzyme linked immunosorbent assay(ELISA)was used to detect the levels of reactive oxygen species(ROS),superoxide dismutase(SOD)and malondialdehyde(MDA),the flow cytometry was used to detect the ratio of apoptosis,Western blot was used to detect Bmal1 and Beclin-1 expression,and the electron microscope was taken to observe the number of autophagosome.Results Compared with NG group,SOD level and Bmal1 expression were decreased in NH/R group,while LDH,ROS,MDA,apoptosis,Beclin-1 and autophagosome were increased(all P<0.05).Compared with NG group,SOD level,Bmal1,Beclin-1 expression and autophagosome were decreased in HG group,while LDH,ROS,MDA,and apoptosis were increased(all P<0.05).Compared with NH/R group,SOD level,Bmal1,Beclin-1 expression and autophagosome were decreased in HH/R group,while LDH,ROS,MDA,apoptosis were increased in HH/R group(P<0.05).Compared with HG group,SOD level and Bmal1 expression were decreased in HH/R group,while LDH,ROS,MDA,apoptosis were increased(all P<0.05),and Beclin-1 and autophagosome showed no difference(P>0.05).Conclusion Oxidative stress induced by high glucose can inhibit the expression of Baml1,which leads to reduce the function of autophagy and aggravate the hypoxia reoxygenation injury.

关 键 词：高糖 氧化应激 时钟基因Bmall 自噬 缺氧复氧 

分 类 号：R363[医药卫生—病理学]