摘要
目的研究生物钟基因芳香烃受体核转位蛋白样1基因(Bmal1)低表达对大鼠胰岛细胞瘤细胞-1(INS-1)功能的影响。方法应用大鼠胰岛β细胞系INS-1细胞,随机将INS-1细胞分为空白组、阴性对照组和Bmal1 siRNA组,通过RNA干扰技术沉默Bmal1,比较三组细胞内活性氧(ROS)水平、胰岛素(GSIS)含量、核因子E2相关因子2(Nrf2)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)mRNA表达,并采用透射电镜观察线粒体结构变化。结果阴性对照组与空白组ROS水平、GSIS含量、Nrf2、CAT、GSH-Px mRNA表达比较,差异无统计学意义(P>0.05);Bmal1 siRNA组ROS水平、Nrf2表达高于阴性对照组,GSIS含量、CAT、GSH-Px mRNA表达低于阴性对照组,差异有统计学意义(P<0.05)。透射电镜结果显示,在空白组和阴性对照组中线粒体为圆形或椭圆形,具有完整的膜和少量管状嵴;Bmal1 siRNA组中线粒体肿胀、形状不规则,线粒体嵴断裂和消失。结论Bmal1低表达可导致β细胞氧化应激增强,GSIS下降,其作用机制可能与影响Nrf2信号通路有关。
Objective To investigate the effect of brain and muscle arnt-like 1(Bmal1)on the function of insulinoma cell line(INS-1 cells).Methods INS-1 cells were randomly divided into blank control group,negative control group and Bmal1 siRNA group.Bmal1 was silenced by RNA interference technology,and the intracellular reactive oxygen species(ROS)levels,glucose-stimulated insulin secretion(GSIS),nuclear factor erythroid-2 related factor 2(Nrf2),catalase(CAT)and glutathione peroxidase(GSH-Px)mRNA expressions were compared in the three groups.Mitochondrial morphological changes were observed by transmission electron microscopy.Results There was no significant difference in the ROS level,GSIS,Nrf2,CAT,GSH-Px mRNA expression between the negative control group and the blank group(P>0.05).The Bmal1 siRNA group had higher ROS levels and Nrf2 expression than the negative control group,and the GSIS,CAT,and GSH-Px mRNA expression were lower than the negative control group.The differences were statistically significant(P<0.05).Transmission electron microscopy results showed that the mitochondria were round or oval in the blank group and the negative control group,with a complete membrane and a few cristae.In the Bmal1 siRNA group,the mitochondria were swollen with irregular shape,and the mitochondrial cristae were fragmented and disappeared.Conclusion Bmal1 inhibition can lead to increased oxidative stress inβ-cell and decreased GSIS via Nrf2 signaling pathway.
作者
叶绿
吴华香
许伟红
YE Lyu;WU Hua-xiang;XU Wei-hong(Department of Rheumatology,School of Medicine,The Second Affiliated Hospital of Zhejiang University,Hangzhou 310009,China)
出处
《医学信息》
2020年第3期66-68,共3页
Journal of Medical Information
基金
浙江省自然科学基金(编号:LQ15H070003)。
