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目的 :研究血管紧张素Ⅱ (AngⅡ )对人近端肾小管上皮细胞结缔组织生长因子mR NA表达的影响。方法 :采用体外培养人近端肾小管上皮细胞株 (HPTC) ,分别观察不同浓度(0、10 -9、10 -7、10 -5mol·L-1)AngⅡ处理 48h ,或用AngⅡ (浓度为 10 -7mol·L-1)处理不同时间 (0、12、2 4、48、72h)对HPTC结缔组织生长因子 (CTGF)mRNA表达的影响 ,CTGFmRNA表达采用逆转录 聚合酶链反应 (RT PCR)方法测定。结果 :AngⅡ (10 -7mol·L-1)作用于HPTC12h后 ,细胞CTGFmRNA的表达开始增加 ,72h达最高水平 ,与对照组相比 ,AngⅡ作用 48h后显著增加〔(0 0 97± 0 0 15 )vs(0 63 2± 0 0 41) ,P <0 0 1〕 ;无AngⅡ的DMEM培养HPTC ,72h内细胞CTGFmRNA水平未见明显改变 (P >0 0 5 )。不同浓度 (10 -9、10 -7、10 -5mol·L-1)AngⅡ作用于HPTC 48h ,CTGFmRNA的表达均有增加 ,分别是对照组 (0mol·L-1)的 1 5、5 5和 7 4倍 (P <0 0 1) ,对扩增产物进行测序证实与基因库中人CTGF的cDNA序列一致。结论 :AngⅡ呈时间和剂量依赖性地刺激HPTCCTGFmRNA表达 ,此结果提示AngⅡ可能通过促进CTGF表达而参与小管间质纤维化的发生
Objective: To investigate the effect of Ang Ⅱ on the expression of connective tissue growth factor mR NA in proximal human proximal tubular epithelial cells. Methods: Human proximal tubular epithelial cell line (HPTC) was cultured in vitro and treated with AngⅡ (0, 10 -9, 10 -7, 10 -5 mol·L -1) for 48 h or with AngⅡ 10 -7 mol·L -1) for different time (0,12,2 4,48,72h) on the expression of connective tissue growth factor (CTGF) mRNA in HPTC. The expression of CTGF mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) Method determination. Results: The expression of CTGF mRNA began to increase after treated with AngⅡ (10 -7 mol·L-1) for 12 h, reaching the highest level at 72 h. Compared with the control group, AngⅡ increased significantly after 48 h [(0 0 97 ± 0 0 15 ) vs (0 63 2 ± 0 0 41), P 0 01). No significant changes in CTGF mRNA level were detected in HPTC cultured with AngⅡ in 72 h (P> 0.05). The expressions of CTGF mRNA in HPTC treated with AngⅡ at different concentrations (10 -9, 10 -7, 10 -5 mol·L -1) for 48 h increased the levels of CTGF mRNA in the control group (0 mol·L -1) 7 4-fold (P <0.01). The amplified products were sequenced and confirmed to be consistent with the cDNA sequence of human CTGF in the gene pool. Conclusion: Ang Ⅱ stimulates the expression of HPTCCTGF mRNA in a time and dose-dependent manner. This result suggests that AngⅡ may be involved in the development of tubulointerstitial fibrosis by promoting the expression of CTGF