CAJ | 학술논문

目的观察嘌呤霉素(PAN)和地塞米松(DEX)干预后足细胞形态及nephrin mRNA表达变化,探讨DEX保护PAN诱导体外培养足细胞损伤的作用机制.方法将体外培养的小鼠肾小球足细胞分为三组,分别为对照组、PAN组和DEX组.对照组加入等体积RPMI-1640培养液培养；PAN组加入PAN(终浓度50 mg/L)；DEX组同时加入PAN(终浓度50 mg/L)和DEX(终浓度1 mmol/L).培养8h、24 h和48 h后,相差显微镜下观察足细胞形态变化；用图像处理软件分析各组细胞胞体面积的差异.采用实时荧光定量聚合酶链式反应检测培养24h、48 h时足细胞nephrin mRNA表达.结果对照组足细胞形态正常,细胞胞体较大,可见树枝样突起及细胞间形成相互连接；PAN诱导足细胞损伤8h、24 h和48 h时,足细胞面积均较对照组明显缩小,分别为对照组的75％、46％和27％(P＜0.01),足细胞足突及细胞间连接消失.DEX组在干预8h、24 h和48 h时,足细胞胞体面积均明显大于PAN组(P＜0.01),部分足细胞可见足突及细胞间连接.PAN组足细胞培养24 h和48 h时,nephrin mRNA表达较对照组明显下降(P＜ 0.01)；DEX组干预后24h及48 h,足细胞nephrin mRNA表达均较PAN组显著升高(P＜ 0.01).结论 DEX对PAN诱导足细胞损伤具有保护作用,其作用机制可能与上调足细胞分子nephrin mRNA表达有关.
목적 관찰표령매소(PAN)화지새미송(DEX)간예후족세포형태급nephrin mRNA표체변화,탐토DEX보호PAN유도체외배양족세포손상적작용궤제.방법 장체외배양적소서신소구족세포분위삼조,분별위대조조、PAN조화DEX조.대조조가입등체적RPMI-1640배양액배양；PAN조가입PAN(종농도50 mg/L)；DEX조동시가입PAN(종농도50 mg/L)화DEX(종농도1 mmol/L).배양8h、24 h화48 h후,상차현미경하관찰족세포형태변화；용도상처리연건분석각조세포포체면적적차이.채용실시형광정량취합매련식반응검측배양24h、48 h시족세포nephrin mRNA표체.결과 대조조족세포형태정상,세포포체교대,가견수지양돌기급세포간형성상호련접；PAN유도족세포손상8h、24 h화48 h시,족세포면적균교대조조명현축소,분별위대조조적75％、46％화27％(P＜0.01),족세포족돌급세포간련접소실.DEX조재간예8h、24 h화48 h시,족세포포체면적균명현대우PAN조(P＜0.01),부분족세포가견족돌급세포간련접.PAN조족세포배양24 h화48 h시,nephrin mRNA표체교대조조명현하강(P＜ 0.01)；DEX조간예후24h급48 h,족세포nephrin mRNA표체균교PAN조현저승고(P＜ 0.01).결론 DEX대PAN유도족세포손상구유보호작용,기작용궤제가능여상조족세포분자nephrin mRNA표체유관.
Objective To observe the changes in podocyte morphology and expression of nephrin mRNA after intervention with pummyein aminonueleoside ( PAN )and then dexamethasone ( DEX ),and to explore the mechanism of action of of DEX in protection of impaired podocytes caused by PAN.Methods The cultured mouse podocytes were divided into 3 groups:control group ( application of RPMI-1640 with the same volume ),PAN group ( PAN of 50mg/L ),and DEX group ( PAN of 50 mg/L plus DEX of 1 μmol/L ).Podocyte morphology was observed with phasecontrast microscopy and analyzed with the image software 8 h,24 h,and 48h after treatment; expression of nephrin mRNA was detected by semi-quantitative RT-PCR 24 h and 48h after treatment.Results In the control group normal podocytes with a larger size, ‘branch-shaped’, and intercellular junction were observed.As compared with the control group,treatment with PAN induced significant shrinkage of podocytes,and the cell size decreased to 75％,46％％,and 27％ at 8 h,24 h,and 48 h,respectively ( P＜ 0.01 ),with disappearance of podocyte foot process and intercellular junction.As compared with PAN group,DEX improved the shrinkage ofpodcytes,and the cell size increased obviously 8 h,24 h,and 48h after treatment( P＜ 0.01 ).RT- PCR revealed the expression of nephrin mR NA decreased more significantly in PAN group than in control group at 24 h and 48 h( P＜ 0.01 ).As compared with PAN group,the expression of nephrin mRNA in DEX group was significantly increased at 24 h and 48 h ( P＜ 0.01 ).Conclusions Dexamethasone has a protective effect on impaired podocytes caused by pummyein aminonueleoside,whose mechanism of action was associated with the upregulation of nephrin mRNA expression in the cells.