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1、白蛋白对近端肾小管上皮细胞增殖与凋亡的影响论文孙良忠,岳智慧,陈述枚【摘要】【目的】探讨白蛋白对近端肾小管上皮细胞(PTCs)增殖与凋亡的影响。【方法】用不同剂量(0.1,0.5,1,5,10,30mg/mL)去脂牛血清白蛋白(dBSA)超载体外培养的NRK52E.freelg/mL作为对照。细胞增殖活性用MTS比色法检测和显微镜观察。细胞凋亡检测:荧光染料DAPI染核,DNA梯带实验,原位末端标记(TUNEL)实验和苔盼蓝染色计数。【结果】高剂量dBSA超载未完全融合NRK52E细胞144h后细
2、胞变得更密集。MTS比色结果发现,高剂量dBSA(5、10、30mg/mL)超载可明显诱导NRK52E细胞增殖,增殖活性分别为(0.700±0.045)A、(1.021±0.029)A和(1.273±0.173)A,较对照细胞(0.441±0.020)A显著升高,P均0.05。高剂量dBSA超载的NRK52E细胞,有较多出现核碎裂和核固缩,有较强DNA梯带和原位末端标记信号。苔盼蓝染色计数发现,高剂量dBSA(5、10、30mg/mL)超载NRK52E细胞72h的死亡细胞数(680000±2861
3、8)/mL,(970000±52915)/mL和(1132500±88954)/mL较对照的死亡细胞数[(312500±28395)/mL显著升高,P均0.05;而且死亡细胞数呈剂量依赖性。【结论】白蛋白超载NRK52E细胞既可诱导细胞增殖,又可诱导细胞凋亡。【关键词】白蛋白;肾小管上皮细胞;细胞增殖;细胞凋亡Abstract:【Objective】Tostudytheeffectsofalbuminoverloadonproliferationandapoptosisinproximaltubu
4、larepithelialcells(PTCs).【Methods】Differentconcentrationofdelipidatedbovineserumalbumin(dBSA)rangedfrom0.1-30mg/mLg/mLascontrol.CellproliferationicroscopeandmeasuredbyOneSolutionCellProliferationAssayKit(MTSassay).Apoptosisethods:DNAladderingassay,Ins
5、ituterminaldeoxynucleotidyltransferase(TdT)-mediateddUTPnick-endlabeling(TUNELassay),nucleichangesmunofluorecentmicroscopeafterstainedidino-2-phenylindole)andtotaldeathcellsicroscope.【Results】Underlightmicroscope,sub-confluentNRK52Ecellsbecamemoreconf
6、luentafterdBSAoverload144h.MTSassayrevealedthathigherdosagesofdBSA(5,10,and30mg/mL)overloadinducedsignificantproliferation(0.700±0.045A,1.021±0.029A,and1.273±0.173A,respectively)48hourslaterinNRK52E,andg/mL)overloadNRK52Ecells,thereorecondensedorbroke
7、nnuclei,strongerDNAladdersandmorepositivesignalsinTUNELassaythanthoseinlog/mL)overloadNRK52E(680000±28618/mL,970000±52915/mL,and1132500±88954/mL,respectively)L),P0.05;Anddeathcellsincreasedinadose-dependentmannerafterdBSAoverload.【Conclusion】Albuminov
8、erloadcouldinduceproliferationandapoptosisinPTCs.Keyin;proximaltubularepithelialcells;proliferation;apoptosis蛋白尿作为众多肾小球疾病的共同特点,不仅提示肾小球的损害程度,而且可能导致肾小管间质损害,促进疾病的进展1,2。但是蛋白尿引起肾小管间质损害的机理仍未完全明了,目前也缺乏有效的预防和治疗措施3。白蛋白是蛋白尿的主要成分4。早期研究发现,白蛋白超载近端肾小管上皮细胞(p