摘要
目的通过体外实验研究，观察糖基化终末产物（AGEs）对体外培养的人腹膜问皮细胞（HPMC）表达纤维连接蛋白（FN）和结缔组织生长因子（CTGF）的影响。观察金雀异黄素（Gen）对AGEs诱导HPMC的FN和CTGF表达的影响。进一步探讨Gen抑制腹膜透析患者腹膜纤维化的机制。方法（1）分别以不同的浓度的AGEs（0、200、600、1000mg／L）刺激HPMC48h后，用RT-PCR方法检测FN的表达情况。（2）将体外培养的HPMC分为5组：正常组（未加任何刺激因素）、AGEs组（AGEs的终浓度为600mg／L）、低剂量Gen组（AGEs和Gen的终浓度分别为600mg／L、25μmol／L）、中剂量Gen组（AGEs和Gen的终浓度分别为600mg／L、50μmol／L）和高剂量Gen组（AGEs和Gen的终浓度分别为600mg／L、1001umol／L），各组予刺激HPMC48h后，用RT—PCR方法检测细胞内FN和CTGFmRNA的水平．ELISA方法测定细胞内FN和CTGF蛋白的表达。结果（1）AGEs剂量依赖性上调HPMCFNmRNA的水平。（2）与正常对照组比较，AGEs组FN、CTGFmRNA的水平和蛋白的表达明显增加（均P〈0．01）；与AGEs组比较，25μmol／L、50μmol／L、100μmol／L的Gen均能明显抑制AGEs诱导的FN、CTGFmRNA的水平和蛋白的的表达（均P〈0．01）。结论AGEs可诱导体外培养HPMC的FN、CTGF基因和蛋白的高表达；而Gen能抑制AGEs诱导的FN、CTGF基因和蛋白的表达。
Objective To investigate the effect of advanced gtycation end products （AGEs） on expression of connective tissue growth factor （CTGF）.fibronectin （FN） in human peritoneal mesothelial celIs（HPMC）.To observe the effect of genistein on expression of CTGF.FN in HPMC induced by AGEs. Methods HPMC were stimulated with different concentrations of AGEs （0,200,600,1000 mg / L） for 48 hours, the expression of FN were detected by RT-PCR. HPMC were divided into 5 group： （1）con- trol; （2）600mg/L AGEs group; （3）Gen treated group with low concentration,the concentration of AGEs were 600 mg / L and the concentration of Gen are 25 p mol/L;（4）Gen treated group with moderate concentration,the concentration of AGEs were 600 mg / L and the concentration of Gen are 50 μ mol/L; （5）Gen treated group with high concentration,the concentration of Weke are 600 mg /L and the concentration of Gen Were 100 μ mol/L; the expression of mRNA for FN and CTGF were measured by RT-PCR,the ex- pression of FN and CTGF protein were detected by ELISA after 48 hours. Results （1）The expression of FN mRNA in human peritoneal mesothelial cells （HPMC） increased in a dose-dependent manner after induced by AGEs; （2）Compared with control, 600mg/L AGEs promoted markedly the expression of mRNA and protein for FN.CTGF;Compared with AGEs,25μ mol/L. 50μmol/L.1001J mol/L Gen inhibited significantly the expression of mRNA and protein for FN,CTGF. Conclusion AGEs can increase markedly the expression of mRNA and protein for FN.CTGF;however,Gen can inhibite the expression of FN.CTGF mRNA and protein stimulated by AGEs,which implies that Gen probably decreases the accumulation of extracellular matrix（ECM） throngh inhibiting the expression of CTGF and has protential ability of anti- peritoneal fibrosis.
出处
《浙江医学》
CAS
2012年第7期515-518,521,共5页
Zhejiang Medical Journal
基金
浙江省自然科学基金资助项目（Y206936）
关键词
糖基化终末产物
腹膜间皮细胞
结缔组织生长因子
纤维连接蛋白
Advanced glycation end products Human peritoneal mesothelial cells Connective tissue growth factorFibronectin Peritoneal dialysis
