一种新型嘧啶基取代芳基苯酸衍生物对人多囊肾囊肿衬里上皮细胞增殖抑制的实验研究

Effects of A Novel PPARγ Agonist on Cell Proliferation in ADPKD Cystic-lining Epithelial Cells

目的:观察新型PPARγ激动剂DH9对人多囊肾囊肿衬里上皮细胞(WT9-12)增殖的影响。方法:予以不同浓度的DH9及罗格列酮作用WT9-12细胞24、48、72、96 h,MTT法检测细胞增殖情况;流式细胞术观察细胞周期的改变;AnnexinV+PI双染色流式细胞术测定细胞凋亡率;Western blotting分析cyclinA、P21CIP/WAF1、Bax和Bcl-2的表达。结果:DH9抑制细胞增殖作用明显优于罗格列酮(P<0.05),并呈量-效和时-效关系。比较加入PPAR-γ抑制剂GW9662前后细胞增殖抑制作用差异无统计学意义(P>0.05)。DH9可增加P21CIP/WAF1蛋白合成,减少CyclinA蛋白合成,使细胞阻滞在S期。DH9作用72 h后Bcl-2/Bax的比值分别为较对照组(2.19±0.08)显著减少(P<0.05),具有促进细胞凋亡的作用。结论:DH9抑制WT9-12细胞增殖活性明显优于罗格列酮,且这种增殖抑制作用与DH9作用在细胞周期的不同调节点,促进WT9-12细胞凋亡有关。

Objective：To investigate the effects of a novel PPARγ agonist DH9 on the proliferation in human polycystic kidney cyst-lining epithelial cells. Methods：The cyst-lining epithelial cells（WT9-12）from human polycystic kidney were treated with DH9 and/or rosiglitazone for 24、48、72、96 hours with or without GW9662.The proliferation was assessed by MTT.Cell cycles were analyzed by flow cytometry.The cyclinA、P21CIP/WAF1、Bax and Bcl-2 were detected by Western blotting. Results：DH9 could effectively inhibit the proliferation of the cells through S cell cycle.DH9 could downregulate the expression of cyclinA and upregulate the expression of P21CIP/WAF1.DH9 could induce apoptosis through Bcl-2/ Bax pathway. Conclusion：DH9 can more effectively block the proliferation of cyst-lining epithelial cells independent of PPARγ.The effect may be mediated by blocking the cell cycle and induce apotisis.