pRb的磷酸化状态在肾小球系膜细胞增生和凋亡中的作用  

Phosphorylated Level of Retinoblastoma Protein on Proliferation and Apoptosis of Glomerular Mesangial Cell

作　　者：马祖祥[1] 何小解[2] 赵维玲[1] 许自川[2] 李永柏[1] 

机构地区：[1]广东省深圳市儿童医院肾脏免疫科,深圳518026 [2]中南大学湘雅二医院小儿肾脏病研究室,长沙410011

出　　处：《中国中西医结合肾病杂志》2010年第9期774-776,852,共4页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：深圳市科技计划项目(No.JH200507120885A)

摘　　要：目的:探讨视网膜母细胞瘤敏感基因蛋白(Rb蛋白)的磷酸化状态对大鼠肾小球系膜细胞(GMCs)增生和凋亡的影响。方法:将大鼠GMCs分为脂多糖(LPS)组、对照组、维生素(VitE)组、激素组与激素+VitE组共5组分瓶培养,后3组加LPS诱导。实验末,收集培养细胞,涂片,应用免疫组化法检测GMCs的增生细胞核抗原(PCNA)、去磷酸化Rb蛋白(Rb)、磷酸化Rb蛋白(pRb)阳性表达率,TUNEL法检测GMCs凋亡指数(AI)。结果:VitE组、激素组、VitE+激素组培养细胞PCNA阳性率均低于LPS组,VitE+激素组培养细胞PCNA阳性率明显低于激素组。VitE组、激素组、VitE+激素组培养细胞凋亡指数均高于LPS组,VitE+激素组培养细胞凋亡指数明显高于激素组。VitE组、激素组、VitE+激素组培养细胞Rb阳性率均低于LPS组,VitE+激素组培养细胞Rb阳性率明显低于激素组。VitE组、激素组、VitE+激素组培养细胞pRb阳性率均低于LPS组,VitE+激素组培养细胞pRb阳性率均低于激素组。PCNA细胞阳性率与pRb阳性细胞率、Rb阳性细胞率均成正相关;AI与pRb阳性细胞率、Rb阳性细胞率均成负相关。结论:去磷酸化和磷酸化Rb蛋白表达水平增加均促进了肾脏系膜细胞的异常增生,抑制异常增生的系膜细胞凋亡,促进肾脏病的慢性进展。Objective：To investigate the phosphorylated level of retinoblastoma protein（Rb） on proliferation and apoptosis of glomerular mesangial cells（GMCs）.Methods：GMCs was divided into 5 groups to culture：control group,lipopolysaccharide（LPS） group,vitamin E（Vit E） group,dexamethasone（DXM） group,VitE＋DXM group,LPS,LPS＋VitE,LPS＋DXM,LPS＋VitE＋DXM were added to the late 4 groups respectively.After 48 hours culture,the GMCs suspension was collected for detecting the positive rate of proliferating cell nuclear antigen（PCNA）,phosphorylated Rb and non-phosphorylated Rb（immunohistochemistry）,and the rate of effective apoptosis（TUNEL method）.Results：The percentage of PCNA positive cells in Vit E group,DXM group,VitE＋DXM group are significantly lower than that of the LPS group（P0.05）;The percentage of PCNA positive cells in VitE＋DXM group are significantly lower than that of the DXM group（P0.05）.The rate of effective apoptosis in Vit E group,DXM group,VitE＋DXM group are significantly higher than that of the LPS group（P0.05）;The rate of effective apoptosis in VitE＋DXM group are significantly higher than that of the DXM group（P0.05）.The percentage of phosphorylated Rb and non-phosphorylated Rb positive cells in Vit E group,DXM group,VitE＋DXM group are significantly lower than that of the LPS group（P0.05）;The percentage of phosphorylated Rb and non-phosphorylated Rb positive cells in VitE＋DXM group are significantly lower than that of the DXM group（P0.05）.The percentage of phosphorylated Rb and non-phosphorylated Rb positive cells showed positive correlation to the percentage of PCNA positive cells,while negatively correlated to the rate of effective apoptosis.Conclusion：The overexpression of phosphorylated Rb and non-phosphorylated Rb are all enhance the proliferation and inhibit the apoptosis of glomerular mesangial cells.

关 键 词：肾小球 系膜细胞 增生细胞核抗原 RB蛋白 凋亡 

分 类 号：R692.6[医药卫生—泌尿科学]