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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
The effect of specific cyclooxygenase-2 inhibitor celecoxib on podocytes apoptosis induced by puromycin aminonucleoside
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DOI:
作者:
孔维信
作者单位:
上海交通大学医学院苏州九龙医院
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
Podocytes; Apoptosis; Puromycin aminonucleoside; Cyclooxygenase-2; Celecoxib
摘要:
目的:研究特异性环氧合酶-2(cyclooxygenase-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,实验分为4组:正常对照组(CON),嘌呤霉素氨基核苷组(PA),塞来昔布组(CEL)和地塞米松组(DEX)。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 blotting技术检测各组足细胞p53及COX-2的表达。结果:随时间延长,PA诱导的足细胞凋亡逐渐增多,在24h及48h明显。各组细胞caspase-3水平无明显差异。24h及48h PA组p53及COX-2表达均较CON组明显增多(p53表达水平24h PA组1.773±0.448比CON组0.288±0.057,48h PA组2.083±0.520比CON组0.283±0.090,P<0.05;COX2表达水平24h PA组6.577±0.667比CON组0.065±0.027,48h PA组5.346±0.865比CON组0.096±0.092,P<0.05)。DEX组及CEL组足细胞凋亡百分率均较PA组明显减轻(其凋亡百分率24h DEX组3.663±0.38比PA组10.973±1.69,48h DEX组4.370±0.78比PA组13.777±1.47,P<0.05;24h CEL组4.080±1.20比PA组10.973±1.69,48h CEL组4.937±0.10比PA组13.777±1.47,P<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有p53、COX-2的表达增加,足细胞凋亡过程与caspase-3无关。使用COX-2抑制剂有抑制足细胞凋亡的作用,作用与地塞米松相当。
Objective: To study the effect of specific cyclooxygenase-2 inhibitor celecoxib on podocyte apoptosis induced by puromycin aminonucleoside and to investigate the protective effect of specific cyclooxygenase-2 inhibitor on podocytes. Method: The conditionally immortalized mouse podocytes were studied and divided into four groups: Control (CON), puromycin aminonucleoside (PA), celecoxib (CEL) and dexamethasone (DEX). The proliferation activity and apoptosis level of podocytes were tested at 0、8、24 and 48 hour respectively. The activity of caspase-3 in apoptotic podocytes was detected and the expressions of p53 and cyclooxygenase-2 in apoptotic podocytes were tested by western blot analysis respectively. Results: The apoptosis of podocytes induced by puromycin aminonucleoside increased over time. The most significance level was found at 24 and 48 hour. The caspase-3 activity has no significant changes in each group. The expressions of p53 and cyclooxygenase-2 were increased significantly after incubated with PA, and the peak was found at 24h and 48h(p53 at 24h PA group 1.773±0.448 vs CON group 0.288±0.057,at 48h PA group 2.083±0.520 vs CON group 0.283±0.090, P<0.05; cyclooxygenase-2 at 24h PA group 6.577±0.667 vs CON group 0.065±0.027,at 48h PA group 5.346±0.865 vs CON group 0.096±0.092, P<0.05 respectively). The podocytes decreased significantly after incubated with both dexamethasone and COX-2 selective inhibitor celecoxib (apoptotic rate at 24hDEX group 3.663±0.38 vs PA group 10.973±1.69,at 48h DEX group 4.370±0.78vs PA group 13.777±1.47,P<0.05;at 24h CEL group 0.639±0.265 vs PA group 6.577±0.667,at 48h CEL group 0.900±0.463 vs PA group 5.346±0.865, P<0.05 respectively). Conclusions: Podocyte apoptosis was induced by puromycin aminonucleoside in a time-dependent manner. The apoptosis increased with the extending of incubating time. The expressions of p53 and cyclooxygenase-2 were increased in apoptotic podocytes induced by PA. Podocyte apoptosis was caspase-3 independent. The specific cyclooxygenase-2 inhibitor celecoxib can inhibit podocyte apoptosis and its effect is comparable to that of dexamethasone.
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