白藜芦醇诱导大鼠原代脂肪细胞凋亡及机制  

Apoptotic Effect and Mechanism of Resveratrol in Rat Primary Adipocytes

作　　者：陈思凡[1] 周妮曼[2] 郑琳[1] 柯梁汝[2] 单智铭[2] 冯翔[1] 

机构地区：[1]中山大学公共卫生学院,广东广州510080 [2]中山大学中山医学院,广东广州510080

出　　处：《中山大学学报（医学科学版）》2010年第5期619-623,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省医学科研基金(A2010143)

摘　　要：【目的】探讨白藜芦醇(Res)对大鼠原代脂肪细胞凋亡的影响及其机制。【方法】培养大鼠原代脂肪细胞,添加不同剂量的Res干预,用Hoechst 33258染色,在荧光显微镜下观察细胞核形态;用DNA Ladder实验观察DNA断裂的条带;检测各组培养基和细胞内的乳酸脱氢酶(LDH)含量,计算LDH漏出率;用流式细胞仪测定细胞凋亡率;Western blot检测相关位点,包括沉默信息调节因子1(Sirt1)、细胞色素C(Cytochrome C)、半胱天冬氨酸酶家族(Caspase 9和Caspase 3)的蛋白质表达。【结果】经过Res干预后,细胞核染色质高度凝集,电泳后出现180～200bp整数倍的寡核苷酸片段。细胞LDH漏出率升高,呈剂量-反应关系(P<0.05或P<0.01)。Res作用后细胞凋亡率明显上升,呈剂量-反应关系(P<0.01)。Sirt1、Cytochrome C、活化的Caspase 9及Caspase 3表达水平上升,胞浆Cytochrome C释放也增加。【结论】Res促进原代脂肪细胞凋亡,其机制可能是通过增加Sirt1表达,经由线粒体凋亡途径,作用于Caspase 9和Caspase 3。Objective To explore the effect and mechanism of resveratrol（Res） on apoptosis of rat primary adipocytes.Methods Rat primary adipocytes were cultured and treated with resveratrol in different dosages.Adipocytes were stained by Hoechst 33258,and cell morphological transformation was examined with fluorescent microscope.DNA Ladder assay was used to further confirm cell apoptosis.The LDH content in medium and cells was measured to calculate the LDH leaking ratio.The number of apoptotic cells were assayed with flow cytometry.The expression of silent information regulator 1（Sirt1）,Cytochrome C,Cleaved Caspase 9,Cleaved Caspase 3 were examined using Western blot.Results Res induced the apoptosis of rat primary adipocytes in a dose dependent manner.Nuclear fragmentation and condensation were observed by Hoechst 33258 staining and DNA Ladder assay.Res also increased the LDH leaking ratio,the number of apoptotic cells and the expression levels of Sirt1 and Cytochrome C,Cleaved Caspase 9,Cleaved Caspase 3.Conclusion Res could induce the apoptosis of rat primary adipocytes.The underlying mechanisms may be enhancing expression of Sirt1,Cytochrome C,Cleaved Caspase 9 and Cleaved Caspase 3 which are related to cell apoptosis.

关 键 词：白藜芦醇 凋亡 沉默信息调节因子1 细胞色素C 半胱天冬氨酸酶 

分 类 号：Q5[生物学—生物化学]