姜黄素对急性亚砷酸钠诱发的小鼠胰岛β细胞毒性损伤的影响  

Effect of curcumin on acute sodium arsenite-induced cytotoxic damage in mouse pancreatic β-cells

作　　者：金壮壮 肖盛达 杨蓓[3] JIN Zhuang-zhuang;XIAO Sheng-da;YANG Bei(Seven-Year-Program of First Clinical College,College of Basic Medical Sciences,China Medical University,Shenyang 110122,China;Clinical Medicineof“5+3”Program,College of Basic Medical Sciences,China Medical University,Shenyang 110122,China;Department of Histology and Embryology,College of Basic Medical Sciences,China Medical University,Shenyang 110122,China)

机构地区：[1]中国医科大学七年制第一临床学院,辽宁沈阳110122 [2]中国医科大学“5+3”一体化临床医学专业,辽宁沈阳110122 [3]中国医科大学基础医学院组织与胚胎学教研室,辽宁沈阳110122

出　　处：《解剖科学进展》2022年第2期213-216,220,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81602825);辽宁省教育厅资助项目(LK201658,JCZR2020002);辽宁省大学生创新创业训练计划资助项目(201810159239,201910159194)。

摘　　要：目的 探讨姜黄素对急性亚砷酸钠诱发的小鼠胰岛β细胞MIN6毒性损伤的影响及其相关机制。方法 将小鼠胰岛β细胞MIN6随机分为4组:正常对照组、4μmol/L亚砷酸钠组、5μmol/L姜黄素预处理+4μmol/L亚砷酸钠组、10μmol/L姜黄素预处理+4μmol/L亚砷酸钠组。采用四甲基偶氮唑蓝(MTT)法检测细胞存活率;免疫印迹法检测细胞内活化型半胱天冬酶-3(cleaved-caspase-3)蛋白表达;免疫细胞荧光技术和免疫印迹法(Western blot)检测细胞内核转录因子Nrf2(nuclear factor-erythroid 2-related factor 2)蛋白表达;实时荧光定量RT-PCR(real-time RT-PCR)检测II相解毒酶NAD(P) H:醌氧化还原酶1(NQO1)和抗氧化酶γ-谷氨酰半胱氨酸合成酶催化亚基(GCLC) mRNA的表达。结果 4μmol/L亚砷酸钠明显诱发MIN6细胞毒性损伤,导致细胞存活率下降,细胞内cleaved-caspase-3蛋白表达升高。经5、10μmol/L姜黄素预处理剂量依赖性降低急性亚砷酸钠对MIN6细胞毒性损伤,明显增加细胞内的Nrf2蛋白表达水平,促进Nrf2核转位,显著上调NQO1和GCLC mRNA表达水平。结论 姜黄素通过预先激活Nrf2信号通路,减轻急性亚砷酸钠诱发小鼠胰岛β细胞的毒性损伤。Objective To investigate the effect of curcumin on acute sodium arsenite(NaAsO;)-induced cytotoxic damage in mouse pancreaticβ-cells and its related mechanism.Methods Mouse MIN6 cells were randomly divided into four groups:control group,4μmol/L NaAsO;group,5μmol/L curcumin+4μmol/L NaAsO;group,10μmol/L curcumin+4μmol/L NaAsO;group.Cell viability was measured by MTT assay,cleaved-caspase-3 protein expression was detected by Western blot.Nrf2 protein expression was detected by immunocytofluorescence and Western blot.The expressions of NQO1 and GCLC mRNAs were detected by Real-time RT-PCR.Results 4μmol/L NaAsO;significantly induced the cytotoxicity of MIN6 cells,resulting in the decrease of cell viability and the increase of cleaved-caspase-3protein expression levles.Pretreatment with 5μmol/L or 10μmol/L curcumin markedly reduced the cytotoxicity of NaAsO;to MIN6 cells in a dose-dependent manner,dramatically increased the expression level of Nrf2 protein in cells,promoted Nrf2 nuclear translocation,and up-regulated the levels of NQO1 and GCLC.Conclusion Curcumin pretreatment alleviates acute sodium arsenite-induced cytotoxic damage in mouse pancreaticβ-cells via activating Nrf2 signaling pathway of MIN6 cells.

关 键 词：2型糖尿病 亚砷酸钠 NRF2 姜黄素 毒性损伤 胰岛Β细胞 小鼠 

分 类 号：R995[医药卫生—毒理学]