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作 者:刘春艳[1] 梅长林[2] 袁莉[2] 张懿[2] 付莉莉[2] 蔡厚安[2] 王雪琦[2]
机构地区:[1]大连医科大学附属二院肾内科,辽宁大连116027 [2]第二军医大学上海长征医院肾内科解放军肾脏病研究所,上海200003
出 处:《基础医学与临床》2010年第7期698-702,共5页Basic and Clinical Medicine
基 金:国家科技重大专项课题(2009ZX09102)
摘 要:目的探讨雷帕霉素对常染色体显性多囊肾病囊肿衬里上皮细胞增殖及血管内皮细胞生长因子(VEGF)表达的抑制作用及其机制。方法 MTT法检测WT9-12细胞增殖;流式细胞术检测细胞周期及凋亡;Western blot检测周期相关蛋白(cyclinD、P21)、凋亡相关蛋白(BCL-2/BAX)及VEGF表达。结果雷帕霉素可抑制WT9-12细胞的增殖,使细胞周期停滞在G0/G1期并促进细胞凋亡。雷帕霉素可下调WT9-12细胞cyclinD、上调P21表达,下调BCL-2、上调BAX表达。原代培养的多囊肾囊肿衬里上皮细胞及WT9-12细胞的VEGFmRNA表达明显高于正常肾小管上皮细胞(P<0.05)。雷帕霉素可抑制WT9-12细胞VEGF的表达(P<0.05)。结论雷帕霉素可通过抑制多囊肾囊肿衬里上皮细胞增殖、促进凋亡及降低VEGF表达而减少血管形成,从而抑制多囊肾病进展。Objective To investigate antiproliferative effect and reduction of VEGF expression of rapamycin on autosomal dominant polycystic kidney disease(ADPKD) cystic lining epithelial cells.Methods ADPKD cystic lining immortalized epithelial cells(WT9-12) were stimulated by rapamycin with different concentrations.MTT method was performed to detect proliferation,VEGF mRNA expression was measured by real time PCR.Western blot method was used to detect the protein expression of cyclinD,P21,BCL-2,BAX and VEGF.Results Rapamycin,a mTOR inhibitor,induced cell growth inhibition,apoptosis and a stable arrest of cells in the G0/G1 phase.Rapamycin downregulated cyclinD,BCL-2 and upregulated P21,BAX expressions.In addition,VEGF mRNA in primary cells from ADPKD patients and from WT9-12 cells was increased as compared with normal tubule cells.Rapamycin downregulated VEGF expression in WT9-12 cells.Conclusion Rapamycin induces ADPKD cystic lining epithelial cells growth inhibition,apoptosis and inhibits VEGF expression,which may be one of mechanisms of rapamycin inhibits ADPKD progression.
关 键 词:常染色体显性多囊肾病 雷帕霉素 细胞周期 凋亡 血管内皮生长因子
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