CAJ | 학술논문

目的研究氯化镉(CdCl2)对离体肝细胞线粒体结构及功能的影响.方法从培养肝细胞(WRL-68)中提取线粒体,各组分别用0、1、5、10靘oL/L CdCl2进行处理.采用分光光度法检测线粒体膜通透性转运孔(MPTP)的开放程度;电镜下观察线粒体的形态改变;罗丹明123检测线粒体膜电位(MMP)的变化;测定线粒体内ATP酶LDH、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)的活性以及丙二醛(MDA)的含量.结果随着CdCl2浓度的增加MPTP开放程度增加,并且MPTP的开放可被环孢霉素A(CsA)所抑制;电镜观察发现,线粒体经CdCl2处理后出现不同程度的肿胀变形;CdCl:诱导MMP显著降低;同时引起SOD、GSH-Px、Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶LDH活性趋于下降,5、10靘oL/L CdCl2显著下调了各种酶的活性(P＜0.05);CdCl2处理组MDA含量升高,10靘oL/L CdCl2处理组与对照组比较有统计学意义(P＜0.05).结论 CdCl2能引起离体肝细胞线粒体结构的破坏,并致MPTP开放、MMP下降、线粒体酶活性的改变等与细胞凋亡相关过程的发生.
목적 연구록화력(CdCl2)대리체간세포선립체결구급공능적영향.방법 종배양간세포(WRL-68)중제취선립체,각조분별용0、1、5、10정oL/L CdCl2진행처리.채용분광광도법검측선립체막통투성전운공(MPTP)적개방정도;전경하관찰선립체적형태개변;라단명123검측선립체막전위(MMP)적변화;측정선립체내ATP매LDH、곡광감태과양화물매(GSH-Px)、초양화물기화매(SOD)적활성이급병이철(MDA)적함량.결과 수착CdCl2농도적증가MPTP개방정도증가,병차MPTP적개방가피배포매소A(CsA)소억제;전경관찰발현,선립체경CdCl2처리후출현불동정도적종창변형;CdCl:유도MMP현저강저;동시인기SOD、GSH-Px、Na+-K+-ATP매、Ca2+-Mg2+-ATP매LDH활성추우하강,5、10정oL/L CdCl2현저하조료각충매적활성(P＜0.05);CdCl2처리조MDA함량승고,10정oL/L CdCl2처리조여대조조비교유통계학의의(P＜0.05).결론 CdCl2능인기리체간세포선립체결구적파배,병치MPTP개방、MMP하강、선립체매활성적개변등여세포조망상관과정적발생.
Objective To study the effects of cadmium on the structure and functions of mitochondria in hepatocytes. Methods Mitochondria were isolated from cultured human hepatocytes of the added into the culture medium. Mitochondrial permeability transition pore (MPTP) opening degree was tested by spectrophotometer. Morphologic changes of mitochondria were observed under transmission electron microscope. The activities of Na+-K+ -ATPase, Ca2 + -Mg2 + -ATPase, LDH, superoxide dismutase ( SOD ) ,glutathione peroxidase (GSH-Px), and the contents of malondialdehyde (MDA) were measured.Mitochondria membrane potential (MMP) was monitored by spectrofluorimeter with fluorescence dye Rh-123. Results CdC12 reduced the absorbance of mitochondria, signifying the opening of MPTP,control group (P ＜ 0.05, P ＜ 0.01 ) . The activity levels of ATPase, LDH, SOD, and GSH-Px in mitochondria decreased in the CdCl2 groups ( all P ＜ 0. 05 ) , and the contents of MDA increased in the CdCl2 groups compared with the control group. Conclusion CdCl2 causes destruction of mitochondria structure,opening of MPTP, decrease of MMP, and changes of vitality of mitochondria enzymes that all play important roles in apoptosis of hepatocytes.