栀子苷抗H_2O_2诱导人脐静脉内皮细胞凋亡的实验研究  

Effect and possible mechanism of geniposide on apoptosis of HUVECs induced by H_2O_2

作　　者：李青岭[1] 于超[1] 李春莉[1] 吴建勇[1] 李文明[1] 

机构地区：[1]重庆医科大学生命科学研究院,重庆400016

出　　处：《中国药理学通报》2010年第10期1367-1371,共5页Chinese Pharmacological Bulletin

基　　金：重庆市自然科学基金资助项目(NoCSTC2009BA5083);重庆医科大学重点基金项目(NoXBED200806);重庆医科大学校级课题(NoXBYB2008059)

摘　　要：目的探讨栀子苷对氧化应激诱导人脐静脉内皮细胞(HUVEC)凋亡的抑制作用及可能机制。方法体外培养的HUVEC细胞用不同浓度的栀子苷药物预处理24h后,加入终浓度为500μmol·L-1H2O2氧化损伤12h。荧光染色法观察细胞内DNA损伤情况,流式细胞术检测细胞凋亡率和线粒体膜电位的变化,Western blot检测Bcl-2和Bax蛋白的表达,酶免法测定caspase-3蛋白酶的活性,Real-time PCR检测超氧化物歧化酶(Mn-SOD)和谷胱甘肽过氧化物酶(GPx)的mRNA表达。结果与模型组相比,不同浓度栀子苷(12.5、25、50mg·L-1)可以明显改善H2O2对细胞内的DNA氧化损伤,降低细胞凋亡率,下调Bax蛋白表达,逐步恢复Bcl-2/Bax表达比例和线粒体膜电位的改变,降低caspase-3蛋白酶活性,上调细胞内Mn-SOD和GPx的表达,但对Bcl-2的表达无影响。结论栀子苷可以抑制H2O2诱导的HUVEC细胞凋亡,其机制可能与调节线粒体应激途径和发挥抗氧化损伤功能有关。Aim To investigate the effect and possible mechanism of geniposide on apoptosis of HUVECs induced by H2 O2.Methods HUVECs were pre-incubated with various doses geniposide（12.5,25,50 mg· L -1 ） for 24 h,then exposed to H 2 O2 with concentration of 500μmol·L -1 for 12 h.The morphology changes of chromosome were observed by invert fluorescent microscope.Apoptosis rate and△Ψm were measured byflow cytometry.The protein expressions of Bcl-2 and Bax were determined by Western blot method.The activation of caspase-3 was detected with the caspase-3 activity assay kit.The mRNA expressions of Mn-SOD and GPx were measured by real-time PCR.Results In comparision with the control（cells treated with H 2 O2 but not geniposide） ,the DNA injury of the cells was attenuated,apoptosis rate,△Ψm,protein expression of Bax and activity of caspase-3 decreased,Bcl-2/Bax protein expression rate and expressions of Mn-SOD and GPx increased in a geniposide-dose-dependent manner,but no narked change occurred in Bcl-2 level. Conclusion Geniposide could inhibit the apoptosis of HUVECs induced by H2 O2 which might be correlated with regulation of apoptosis in the path of mitochondrion and regulation of Mn-SOD and GPx expression.

关 键 词：栀子苷 HUVEC细胞 氧化应激 细胞凋亡 线粒体途径 抗过氧化酶 

分 类 号：R284.1[医药卫生—中药学] R322.123[医药卫生—中医学]