CAJ | 학술논문

目的探讨β肾上腺素受体(β-AR)激活对人肾小球系膜细胞(HMC)凋亡的影响及其机制.方法 HMC被分为空白对照(Ctrl)组;β-AR激活(NE/Pra)组;β-AR抑制(Prop)组;抗氧化剂(VC和NAC)组.采用反转录PCR法检测系膜细胞β肾上腺素受体的表达;激光共聚焦术测定细胞内钙信号变化;Tunel法检测细胞凋亡;Western印迹法检测细胞内半胱天冬酶3 (Caspase-3)的表达.结果反转录PCR结果显示,人系膜细胞有β1-AR和β2-ARmRNA表达;激光共聚焦结果显示,β1-AR和β2-AR激动剂均能诱导系膜细胞内钙信号变化(P＜ 0.05);β-AR激活组诱导人系膜细胞产生活性氧类(ROS)增加,β-AR激动剂作用后ROS从0.5 h开始增多,4h达到高峰,之后减弱,但在12h时,细胞内ROS仍然超过对照组(P＜0.01).并且随着β-AR激动剂浓度增加,ROS生成增加.与对照组相比,β-AR激活组诱导人系膜细胞凋亡率增加,并随β-AR激动剂浓度增加和作用时间延长,凋亡细胞数随之增加(均P＜ 0.01).抗氧化剂维生素C和NAC可减轻由β-AR激活诱导的细胞凋亡(P＜0.01),β-AR阻断剂普萘洛尔可减轻细胞内ROS产生和β-AR激活诱导的细胞凋亡.与对照组相比,β-AR激活可诱导系膜细胞Caspase-3表达上调(均P＜ 0.01).结论 β-AR激活可诱导人系膜细胞凋亡,其机制可能是与β-AR激活增加细胞内氧化应激水平有关.
목적 탐토β신상선소수체(β-AR)격활대인신소구계막세포(HMC)조망적영향급기궤제.방법 HMC피분위공백대조(Ctrl)조;β-AR격활(NE/Pra)조;β-AR억제(Prop)조;항양화제(VC화NAC)조.채용반전록PCR법검측계막세포β신상선소수체적표체;격광공취초술측정세포내개신호변화;Tunel법검측세포조망;Western인적법검측세포내반광천동매3 (Caspase-3)적표체.결과 반전록PCR결과현시,인계막세포유β1-AR화β2-ARmRNA표체;격광공취초결과현시,β1-AR화β2-AR격동제균능유도계막세포내개신호변화(P＜ 0.05);β-AR격활조유도인계막세포산생활성양류(ROS)증가,β-AR격동제작용후ROS종0.5 h개시증다,4h체도고봉,지후감약,단재12h시,세포내ROS잉연초과대조조(P＜0.01).병차수착β-AR격동제농도증가,ROS생성증가.여대조조상비,β-AR격활조유도인계막세포조망솔증가,병수β-AR격동제농도증가화작용시간연장,조망세포수수지증가(균P＜ 0.01).항양화제유생소C화NAC가감경유β-AR격활유도적세포조망(P＜0.01),β-AR조단제보내락이가감경세포내ROS산생화β-AR격활유도적세포조망.여대조조상비,β-AR격활가유도계막세포Caspase-3표체상조(균P＜ 0.01).결론 β-AR격활가유도인계막세포조망,기궤제가능시여β-AR격활증가세포내양화응격수평유관.
Objective To investigate the effects of β-adrenoceptor (β-AR) activation on the apoptosis in human mesangial cells and it's mechanism.Methods Cultured HMC were used in experiments and were divied into four groups:the control group; β-AR activation (β-AR agonist NE/Pra) group; β-AR inhibitor (Prop) group; antioxidants group.The experiments technology including PCR,confocal scanning microscope,immunofluorescence and Tunel.Results The results of RTPCR and confocal scanning microscope showed that β1-AR and β2-AR were expressed in human HMC.β-AR activation induced reactive oxygen species (ROS) increase in human MCs,the relative levels of ROS were elevated as early as 0.5 h after β-AR activation,and gradually increased and peaked at 4 h on a concentration and time dependent manner.Tunel results demonstrated that β-AR activation induced apoptosis with ROS on a concentration and time dependent manner,β-AR blocking agent-propranolol significantly inhibited β-AR activation induced apoptosis.Antioxidants including vitamin C and NAC could inhibited β-AR activation induced apoptosis (all P ＜ 0.01).Conclusions β-AR is functionally expressed in human mesangial cell,furthermore β-AR activation-induced ROS increase mediate apoptosis.Antioxidants can inhibit β-AR activation induced apoptosis.