特异性环氧合酶-2抑制剂塞来昔布对嘌呤霉素氨基核苷诱导的足细胞凋亡的影响  

Effect of specific cyclooxygenase-2 inhibitor celecoxib on podocyte apoptosis induced by puromycin aminonucleoside

作　　者：丁露露[1] 孔维信[1] 顾勇[2] 杨海春[2] 陈靖[2] 朱莉[3] 

机构地区：[1]上海交通大学医学院附属苏州九龙医院肾内科,苏州215021 [2]复旦大学附属华山医院肾脏病研究所 [3]江苏大学附属人民医院

出　　处：《上海医学》2011年第4期292-297,242,共6页Shanghai Medical Journal

基　　金：江苏省苏州市科技计划项目资助(SS0716)

摘　　要：目的研究特异性环氧合酶-2(COX-2)抑制剂塞来昔布对嘌呤霉素氨基核苷(puromycin aminonucleoside,PA)诱导的足细胞凋亡的影响,初步探讨特异性COX-2抑制剂对足细胞保护作用的机制。方法以条件永生性小鼠足细胞株为研究对象,分为正常对照(CON)组、PA组、塞来昔布(CEL)组和地塞米松(DEX)组。在0、8、24和48h检测各组足细胞活性和凋亡水平,并检测足细胞凋亡过程中caspase-3的活性水平,运用Western印迹法检测各组足细胞P53及COX-2的表达。结果与CON组比较,PA、CEL、DEX组在24、48h的足细胞凋亡率均显著升高(P值均<0.05),但各组这两个时点间的足细胞调亡率的差异无统计学意义(P值均>0.05)。与PA组比较,CEL、DEX组在24、48h的足细胞凋亡率均显著降低(P值均<0.05)。与CON组比较,各组细胞caspase-3活性无明显变化(P值均>0.05)。与CON组比较,PA组在24、48h的P53、COX-2表达均显著增多(P值均<0.05),各组这两个时间点间P53、COX-2表达的差异无统计学意义(P值均>0.05)。与PA组同时间点比较,CEL、DEX组在24、48h的P53、COX-2的表达显著下降(P值均<0.05)。结论 PA诱导的足细胞凋亡呈时间依赖性,随时间延长,足细胞凋亡逐渐增多。PA诱导的足细胞凋亡过程中P53、COX-2的表达增加,足细胞凋亡过程与caspase-3无关。COX-2抑制剂具有抑制足细胞凋亡的作用,与地塞米松相当。Objective To determine the effect of specific cyclooxygenase （COX）-2 inhibitor celecoxib on podocyte apoptosis induced by puromycin aminonucleoside （PA） and to investigate the protective effect of specific COX-2 inhibitor on podocytes. Methods The conditionally immortalized mouse podocytes were divided into four groups： Control （CON）, PA, celecoxib （CEL） and dexamethasone （DEX）. The proliferation and apoptosis of podocytes were tested by MTT assay and Hoechst 33258 staining at 0, 8, 24 and 48 h after corresponding treatment, respectively. The activity of caspase-3 in apoptotic podocytes was detected with caspase-3 kit and the expressions of P53 and COX-2 in apoptotic podocytes were tested by Western blotting analysis. Results Compared with CON group, the apoptosis of podocytes induced by PA was increased in PA, CEL and DEX groups （P〈0.05）. The most prominent apoptosis was found at 24 and 48 h （P〉0.05）, but there was no difference among the latter 3 groups （P〈0.05）; compared with PA group, those of CEL and DEX groups were obviously decreased （P〈0.05）. The caspase-3 activity had no significant changes in each group in comparison with CON group （P〉0.05）. The expressions of P53 and COX-2 were increased significantly after incubated with PA （P〈0.05）, and the peak was found at 24 and 48 h （P53 at 24 h PA group 1.773±0.448 vs. CON group 0.288±0.057, at 48 h PA group 2.083±0.520 vs. CON group 0.283±0.090, P〈0.05; COX-2 at 24 h PA group 6.577±0.667 vs. CON group 0.065±0.027, at 48 h PA group 5.346±0.865 vs. CON group 0.096±0.092, P〈0.05 respectively）. Their expressions were decreased in CEL and DEX groups after 24 and 48 h treatment when compared with those in PA group （P〈0.05）. Conclusion Podocyte apoptosis shows a time-dependent manner after PA treatment. The expressions of P53 and COX-2 are increased during the process, while caspase-3 activity has no obvious change. Specific COX-2 inhibitor celecoxib exerts inhibitory effect on the apoptos

关 键 词：足细胞 凋亡 嘌呤霉素氨基核苷 环氧合酶-2 塞来昔布 

分 类 号：R692[医药卫生—泌尿科学]