罗格列酮对多囊肾囊肿衬里上皮细胞p38促分裂原活化蛋白激酶信号通路的影响  

Effect of rosiglitazone on p38 mitogen-activated protein kinase pathway in polycystic kidney cyst-lining epithelial cells

作　　者：贾洁爽[1] 梅长林[1] 付莉莉[1] 戴兵[1] 胡惠民[1] 

机构地区：[1]第二军医大学附属长征医院肾内科解放军肾脏病研究所,上海200003

出　　处：《中华肾脏病杂志》2009年第6期452-457,共6页Chinese Journal of Nephrology

基　　金：国家“863”项目（2007AA022321）;国家自然科学基金面上项目（30771016,30570867）

摘　　要：目的探讨罗格列酮对多囊肾囊肿衬里上皮细胞p38促分裂原活化蛋白激酶（MAPK）信号通路的作用。方法分别用罗格列酮（RGZ，10μmol／L）、过氧化物酶体增殖物活化受体γ（PPARγ）抑制剂GW9662（10μmol／L）、RGZ（10μmol／L）＋GW9662（10μmol／L）、p38MAPK特异性抑制剂SB203580（10txmol／L）、SB203580（10μmol／L）＋RGZ（10μmol／L）处理体外培养的多囊肾囊肿衬里上皮细胞（PKD细胞）2h后，再用表皮生长因子（EGF）刺激不同时间，另设置空白对照组和单独EGF刺激组。采用Western印迹方法检测p38MAPK、磷酸化p38MAPK（p-p38）、增殖细胞核抗原（PCNA）表达；RT—PCR检测p38mRNA表达；免疫细胞化学检测c—fns和c-jun的表达。结果（1）与空白对照组相比，EGF显著上调p—p38、PCNA、c—fos和c—jun的表达（P〈0．01）。（2）与EGF单独刺激相比，RGZ显著降低p38活化和基因表达（均P〈0．01）。RGZ组、SB203580＋RGZ组p-p38、PCNA、c—fos、c—jun表达明显下调（P〈0．01），两组间差异无统计学意义。（3）与RGZ组相比，RGZ＋GW9662组部分阻断RGZ的下调作用（P〈0．05）。结论罗格列酮抑制多囊。肾囊肿衬里上皮细胞增殖的作用机制可能与其降低p38MAPK活性，继而抑制PCNA、c—fos及c-jun表达有关。这种抑制作用是部分非PPARγ依赖的。Objective To investigate the effect of rosiglitazone on p38 mitogen-activated protein kinase （p38MAPK） pathway in polycystic kidney cyst-lining epithelial cells. Methods The cyst-lining epithelial cells （PKD cells） from human polycystic kidney were treated with rosiglitazone （10μmol/L）, peroxisome proliferator-activated receptor-γ （PPARγ） inhibitor GW9662 （10 μmol/L）, rosiglitazone （10μmol/L） ＋GW9662 （10μmol/L）, p38MAPK specific inhibitor SB203580 （10 μmol/L）, SB203580 （10μmol/L）＋ rosiglitazone（ 10μmol/L） for 2 hours followed by epidermal growth factor （EGF） stimulation. Protein expressions of p38, phospho-p38 （p-p38） and proliferating cell nuclear antigen （PCNA） were detected by Western blot. p38 mRNA was examined by RT-PCR. Expression of c-fns and c-jun was observed by immunoeytochemistry. Results （1） EGF markedly up-regulated the expressions of p38, p-p38, PCNA, c-los and c-jun compared with control group （P〈0.O1）. （2） Compared with EGF treated group, rosiglitazone significantly reduced p38 activation and mRNA expression （P〈0.01, respectively）. Rosiglitazone, rosiglitazone＋ SB203580 could significantly down-regulated p-p38, PCNA, c-los and c-jun expression （P〈0.01, respectively） with no significant difference between these two groups. （3） GW9662 partially reversed the reduction effect of rosiglitazone. Conclusions Rosiglitazone can inhibit proliferation of autosomal dominant polycystic kidney disease cyst-lining epithelial cells partially through downregulating p38 activation and reducing c-los, c-jun and PCNA expression. The above effect of rosiglitazone is in part PPARγ-independent.

关 键 词：P38丝裂原活化蛋白激酶类 多囊肾 常染色体显性 表皮生长因子 罗格列酮 

分 类 号：R692.1[医药卫生—泌尿科学]