β肾上腺素受体激活诱导人系膜细胞凋亡  

β- adrenoceptor activation induces the apoptosis of human mesangial cells

作　　者：孔凡武[1] 刘玉宝[2] 梁丽杰[2] 刘秋爽[3] 任野平[1] 

机构地区：[1]哈尔滨医科大学附属二院肾内科,150086 [2]齐齐哈尔医学院附属二院ICU [3]哈尔滨医科大学

出　　处：《中华肾脏病杂志》2015年第7期516-520,共5页Chinese Journal of Nephrology

基　　金：黑龙江省卫生计生委科研基金（2014-316）

摘　　要：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-AR mRNA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0．5h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。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 RT- PCR 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.

关 键 词：系膜细胞 交感神经 凋亡 氧化性应激 

分 类 号：R692[医药卫生—泌尿科学]