新型过氧化物酶体增殖物激活受体激动剂对人多囊肾囊肿衬里上皮细胞Wnt-β连环素信号通路的影响  

Effects of a novel PPARy agonist on Wnt-β-catenin pathway epithelial cells

作　　者：付莉莉[1] 刘沫言[2] 刘春艳[1] 胡惠民[1] 梅长林[1] 

机构地区：[1]第二军医大学长征医院肾内科解放军肾脏病研究所,上海200003 [2]济南军区总医院医务部

出　　处：《中华肾脏病杂志》2012年第6期464-468,共5页Chinese Journal of Nephrology

基　　金：上海市重点学科基金（B902）

摘　　要：目的观察新型过氧化物酶体增殖物激活受体（PPARγ）激动剂DH9对人多囊肾囊肿衬里上皮细胞（WT9—12）中Wnt-β连环素（catenin）信号通路的影响。方法予以不同浓度的DH9作用WT9—12细胞72h，MTF法检测细胞增殖情况；RT—PCR技术检测DH9干预后β—cateninmRNA水平。予GSK3β抑制剂（SB216763）或PPARγ抑制剂GW9662预处理WT9-12细胞后，再予以60μmol／LDH9或单给DH9干预，Western印迹分别分析β—catenin、磷酸化β-catenin、GSK3β、磷酸化GSK3β蛋白水平。结果DH9对WT9—12细胞增殖有抑制作用，60μmol／LDH9干预72h对细胞的增殖抑制达50％，并随着干预时间延长，抑制率增高。60μmol／LDH9可下调p—catenin的蛋白表达（P〈0．01），上调磷酸化β-catenin的蛋白表达（P〈0．01），具有剂量依赖性。而DH9对Bcatenin的mRNA水平无明显改变。DH9可上调磷酸化GSK3β的蛋白表达（P〈0．01），但对GSK3β的表达无明显影响。在SB216763预处理WT9—12细胞与DH9共处理72h后，发现可逆转DH9对β—catenin的下调作用（P〈0．01），但GW9662对DH9下调β—catenin的作用无明显改变。结论DH9抑制WT9—12细胞的增殖具有时间和剂量依赖性，并且这种增殖抑制作用可通过GSK313依赖的方式下调β—catenin的蛋白表达从而抑制Wnt-β-catenin信号通路实现，与PPARγ途径无关。Objective To investigate the effects of a novel PPARy agonist DH9 on Wnt- β-catenin pathway in human polycystic kidney cystic-lining epithelial cells （WT9-12）. Methods WT9-12 cells were treated with different concentrations of DH9 for 72 hours and the proliferation was assessed by MTT. W39-12 cells were pretreated with SB216763 or GW9662 for two hours and then treated with DH9 for 72 hours. Western blotting was applied to detect the protein expression of β-catenin, phospho-β-catenin, GSK3β, phospho-GSK3β. Results DH9 could effectively inhibit the proliferation of the cells. 60 μmol/L DH9 could facilitate β-catenin down-regulation （P〈 0.01） and phospho-β-catenin up-regulation （P〈0.01）. Inhibition of GSK3β by SB216763 could protect WT9-12 cells against DH9-facilitated β-catenin repression in a dose-dependent manner despite phosphorylating deactivation, but PPARy inhibitor GW9662 couldn＇t. Conclusions DH9 can effectively block the proliferation of WTg-12 cells. The effect may be mediated by facilitating the down-regulation of β-catenin via GSK3β-dependent mechanism.

关 键 词：过氧化物酶体增殖物激活受体 多囊肾 常染色体显性 细胞增殖 多囊肾囊肿衬里上皮细胞 Wnt-β连环素 

分 类 号：R692.1[医药卫生—泌尿科学]