Objective To investigate the role of cyclophilin D ( Cyp D ) on oxidative stress -induced necrosis in renal tubular epithelial cells .Methods Eighteen male Wistar rats were randomly divided into Control group , Sham con-trol group and Ischemia reperfusion injury group .HE staining was carried out and serum creatinine was measured .H2 O2 was used to induce oxidative stress by different times or concentrations .Cyp D-siRNA or CsA were applied as pretreat-ment .The manner of cell death was identified by flow cytometry .Results Serum creatinine was significantly increased in Ischemia reperfusion injury group than those in Control group and Sham control group (P=0.017, P=0.018).Proximal renal tubulars expansion , loss of brush border , and a large number of tube type and cell vacuoles degeneration were ob-served in Ischemia reperfusion injury group by HE staining .H2 O2 induced HK-2 cells necrosis in dose -and time-de-pendent manners, which was inhibited by Cyp D -siRNA and CsA (P=0.026, P=0.008).Conclusion Cyp D plays an important role in oxidative stress -induced necrosis , which could be ameliorated by inhibiting Cyp D .%目的 探讨亲环素D(Cyp D)在氧化应激时对肾小管上皮细胞坏死的影响.方法 Wistar雄性大鼠随机分对照组(n=6)、假手术组(n=6)及缺血再灌注组(n=6),观察各组血肌酐及肾小管坏死情况.体外培养人近端肾小管上皮细胞HK-2细胞系,以不同浓度(0.75、1.5、3 mmol/L)过氧化氢作用12 h及以1.5 mmol/L过氧化氢作用HK-2细胞不同时间(6、12、24 h),观察不同处理条件下细胞的坏死情况;使用环孢素A(CsA)或Cyp D-siRNA预处理、1.5 mmol/L过氧化氢作用HK-2细胞12 h后,观察细胞坏死情况.结果 缺血再灌注组大鼠的血肌酐较对照组及假手术组明显升高(P=0.017,P=0.018);肾组织行HE染色,可见缺血再灌注组近端肾小管明显扩张、刷状缘消失,并存在大量管型、细胞空泡变性以及上皮细胞坏死脱落,对照组和假手术组未见肾脏组织损伤.H2 O2刺激HK-2细胞系,细胞坏死呈时间及浓度依赖性,随着作用时间增加及H2 O2浓度的增加,HK-2细胞坏死率显著增加;Cyp D-siRNA及Cyp D抑制剂CsA均可明显减轻H2 O2所诱导的细胞坏死(P=0.026,P=0.008).结论 Cyp D介导了氧化应激诱发的肾小管上皮细胞坏死,通过抑制Cyp D可以改善氧化应激下肾小管上皮细胞的存活率,从而在一定程度上减少缺血再灌注诱发的急性肾损伤.
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