酞菁锌光动力治疗诱导大肠癌Lovo细胞产生活性氧  

Phthalocyanine zinc photodynamic therapy-induced ROS of Lovo cells

作　　者：李涛[1] 王玉[1] 陈伟[2] 夏春辉[1] 伦志强 LI Tao;WANG Yu;CHEN Wei;XIA Chun-hui;LUN Zhi-qiang(Department of Basic Medicine,Qiqihar Medical University,Qiqihar 161006,China;College of Chemistry and Chemical Engineering,Qiqihar University)

机构地区：[1]齐齐哈尔医学院基础医学院,161006 [2]齐齐哈尔大学化学与化工学院

出　　处：《天津医药》2020年第4期253-257,I0001,共6页Tianjin Medical Journal

基　　金：黑龙江省教育厅资助项目(12541908)。

摘　　要：目的探究P38MAPK介导四-α-(对羧基苯氧基)酞菁锌(TαPcZn)光动力治疗(PDT)诱导的人结肠癌Lovo细胞线粒体损伤的机制.方法将细胞分别设立对照组、siRNA-阴性对照组、siRNA-P38MAPK转染组、TαPcZn-PDT组和TαPcZn-PDT/siRNA-P38MAPK组,各组细胞处理后经红光照射10 min,然后孵育3 h.孵育结束后RT-PCR和Western blot检测P38MAPK沉默效果.采用DCFH-DA探针检测各组细胞活性氧(ROS)水平、JC-I/7-AAD双标染色法检测线粒体膜电位(ΔΨm)的变化,AnnexinV-FLOUS/7-AAD双标染色法检测细胞凋亡率.结果siRNA干扰后P38MAPK的mRNA和蛋白表达水平明显下降.TαPcZn-PDT明显诱导Lovo细胞ROS增加,引起线粒体ΔΨm去极化,细胞凋亡率升高.siRNA沉默P38MAPK后,TαPcZn-PDT诱导Lovo细胞产生的ROS减少,发生ΔΨm去极化的细胞减少,凋亡率下降.结论在TαPcZn-PDT作用下,沉默P38MAPK基因可明显抑制ROS的产生及释放,减弱线粒体ΔΨm的去极化,从而降低光激发TαPcZn所诱导的细胞凋亡.Objective To investigate the P38MAPK-mediated mitochondrial damage induced by TαPcZn-PDT in Lovo cells.Methods Lovo cells were divided into control group,siRNA-negative group,siRNA-P38MAPK group,TαPcZn-PDT group and TαPcZn-PDT/siRNA-P38MAPK group.The cells in each group were treated and irradiated by red light for 10 minutes and then incubated for 3 hours.After incubation,RT-PCR and Western blot assay were used to detect the effect of p38MAPK silencing.The cell ROS levels were detected by DCFH-DA probe.JC-I/7-AAD double-labeled staining was used to detect changes of cell mitochondrial membrane potential(ΔΨm).Annexin V-FlOUS/7-AAD double-labeled staining was used to detect the cell apoptotic rate.Results After siRNA interference,the protein expressions of P38MAPK mRNA significantly decreased.TαPcZn-PDT induced the increase of ROS,the depolarization of mitochondrialΔΨm and the apoptotic rate.After siRNA silencing P38MAPK,the ROS was decreased,the number of depolarized cells was decreased and the apoptosis was weakened,which was induced by TαPcZn-PDT in Lovo cells.Conclusion That silencing of P38MAPK can significantly inhibit the production and release of ROS and decrease the depolarization of mitochondrialΔΨm,thus attenuate the apoptosis induced by TαPcZn-PDT.

关 键 词：活性氧 膜电位 线粒体 细胞凋亡 丝裂原激活蛋白激酶 P38MAPK 酞菁锌 

分 类 号：R349.5[医药卫生—基础医学]