摘要
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 。
Objective To observe the effect of angiotensin Ⅱ(Ang Ⅱ) on the expression of CTGF mRNA in cultured HPTC.Methods The cell line of HPTC were cultured in Dulbeccos modified Eagle medium (DMEM) containing 10% fetal calf serum.After incubation with serum free DMEM for 24 h,cells were incubated in absence or prescence of various concentrations(0,10 -9,10 -7,10 -5 mol·L -1) of Ang Ⅱ for 48 h,or incubated with Ang Ⅱ(10 -7 mol·L -1) for different period (12?h,24?h,48?h,72?h).Total RNA was isolated from cultured HPTC by TRIZOL method.RT?PCR was performed using access RT?PCR kit.The reaction products were measured by 2% agarose gel electrophoresis.All experiments were performed in triplicate.Results CTGF mRNA expression was elevated 12 h after cells exposed to 10 -7?mol·L -1 Ang Ⅱ and reached its peak level at the end of 72 h.Compared to control group,it increased to 6.3 folds after 48 h incubation(0.097±0.015 vs 0.631?7±0.041,P<0.01).When different concentration(0,10 -9,10 -7,10 -5mol·L -1) of Ang Ⅱ were added to the medium for 48 h,CTGF mRNA expression increased 1.5, 5.5 and 7.4 folds respectively (P<0.01).The consequence analysis for the PCR products demonstrated the same as human CTGF cDNA in gene bank.Conclusion Ang Ⅱ induced a time-and dose-dependent expression of CTGF mRNA in HPTC,which implied a profibrotic role of Ang Ⅱ in mediating tubulointerstitial fibrosis possibly through stimulating the production of CTGF.
出处
《东南大学学报（医学版）》
CAS
2003年第2期84-88,共5页
Journal of Southeast University(Medical Science Edition)
基金
江苏省自然科学金资助项目(98 BS 36)
江苏省医学重点人才基金()资助项目
