摘要
目的研究小剂最环孢霉素A(CsA)对大鼠缺血性急性肾衰竭诱导的肾小管上皮细胞线粒体通透性转运的影响,探讨其抑制肾小管上皮细胞凋亡的作用机制。方法Wistar大鼠双侧肾动脉夹闭30min诱导缺血性急性肾衰竭。再灌注18h后留取肾组织及血液标本。比色法测定血清肌酐(SCr)浓度,原位末端标记(TUNEL)法检测肾小管上皮细胞凋亡情况,Westernblot测定肾组织中胞浆细胞色素C、caspase-3、9蛋白质表达。结果肾脏缺血30min/再灌注18h后大鼠SCr明显升高[(106.9±19.4)μmol/Lvs(56.5±7.1)μmol/L,P<0.05],肾小管上皮细胞凋亡数目显著增加[(20.14±3.70)%vs(0.99±0.17)%,P<0.05],肾组织胞浆细胞色素c、caspase-3、9表达增加(P<0.05);CsA能显著降低SCr[(87.5±18.5)μmol/L],减轻肾小管上皮细胞凋亡[(14.61±3.39)%],下调胞浆细胞色素C、caspase-3、9表达(P<0.05)。结论小剂量CsA能抑制大鼠缺血性急性肾衰竭引起的线粒体通透性转运,从而减轻肾小管上皮细胞凋亡,保护肾功能。
Objective To investigate the effect of low dose cyclosporine A (CsA) on mitochondrial permeability transition of tubular cells induced by ischemic acute renal failure (ARF) in a rat model. Methods Male Wistar rats were divided randomly into 3 groups (sham, ARF and ARF+CsA). ARF was induced by clamping both renal arteries for 30 min. The animals were pretreated with vehicle or CsA intraperitoneally 15 min before ischemia, and were sacrificed at 18h of reperfusion. Renal function following ARF was determined by measuring serum creatinine. Tubular cell apoptosis was confirmed by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) assay. Protein expressions of renal cytosolic cytochrome c, caspase-3, 9 were analyzed by Western blot analysis. Results Serum creatinine levels significantly increased after acute renal injury [(106.9±19.4)μmol/L vs (56.5±7.1)μmol/L, P〈0.05]. Renal ischemia injury induced a significant increase of apoptotic tubular cells [(20.14±3.70) % vs (0.99±0.17)%, P〈0.051]. Protein expressions of cytosolic cytochrome c, caspase-3, 9 in the kidneys of ARF group were markedly up-regulated (P〈0.05). CsA significantly improved renal funetion[(87.5±18.5)μmol/L], reduced apoptotic tubular ceils and inhibited the up-regulation of cytosolic cytochrome c, easpase-3, 9 expressions (P〈0.05). Conclusions Low-dose CsA inhibits mitochondrial permeability transition of tubular cells in ischemic ARF, which may essentially contribute to its anti-apoptotic effects.
出处
《中国血液净化》
2005年第11期613-617,共5页
Chinese Journal of Blood Purification
基金
国家重点基础研究发展规划"973"资助项目(G2000057000)
国家自然科学基金创新研究群体项目(30121005)
关键词
环孢霉素A
肾脏
急性肾衰竭
线粒体
通透性转运
Cyclosporine A Renal
Acute renal failure
Mitochondria
Permeability transition