GRβ对肾小球系膜细胞糖皮质激素效应的影响.pdfVIP

中南大学学报‘ 学版 J Cent South Univ Med Sci( ) 2007,32 ) 。 (6) … 94l ① Effect of gluc0c0rtic0id receptor beta on glucocorticoid action in glomerular mesangial cells ZHANG Lei ，HE Qing-nan ，ZHU Min ，ZHOU Gang ， DING Juan．juan ，ZHOU Pin ，WU Xiao．chuan ，YI Zhu．wen (1．Laboratory ofPediatric Nephrology，Second Xiangya Hospital，Central South University，Changsha 41O01 1； 2．Molecular Biology Research Center，Xiangya School ofMedicine，Central South University，Changsha 41O078，China) Abstract： Objective To construct mesangial cell lines over-or under-expressing glucocorticoid receptor B(GR[3)，to investigate the effect of GR[3 on glucocorticoid biological function，and to deter- mine whether the overexpression of GR[3 explains the glucocorticoid-resistant in glomerular mesangial cells(GMCs)．Methods The recombinant human sense or anti-sense gene of GR[3 was transferred into the rat GMCs by retrovirus-mediated stable transfection technique．Expression of hGR[3 mRNA in GMCs was determined by reverse transcription of total RNA followed by quantitative reverse transcription- polymerase chain reaction(RT-PCR)，and the product of RT-PCR was then analyzed by gene sequen- cing．The expression of hGR[3 protein in GMCs was tested by Western blot．The inhibitory rate of dexa- methasone．mediated cells on lip0lysaccharide(LPS)-stimulated GMC proliferation was detected to as- sess the effect of GRB at different expression levels on the glucocorticoid action．The cell proliferative activity in different cells with different levels of GR[3 was tested by MTr chromatometry．The change of cell cycle was analyzed by flow cytometry．Results RT-PCR and gene sequencing show