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作 者:郝志宏[1] 于力[1] 王丽娜[1] 温捷[1] 张瑶[1]
机构地区:[1]广州医学院附属广州市第一人民医院儿科,广东广州510180
出 处:《临床儿科杂志》2012年第4期362-366,共5页Journal of Clinical Pediatrics
基 金:广州市科技计划项目(No.2010Y1-C521)
摘 要:目的观察福辛普利(fosinopril,FOS)对脂多糖(LPS)诱导的大鼠肾小球系膜细胞(GMC)β1整合素表达及分泌细胞外基质(ECM)的影响。方法大鼠GMC原代体外培养,鉴定3~10代后用于实验。实验分为5组:对照组、LPS刺激组(LPS)、FOS高、中、低剂量组(分别为FOS1组、FOS2组、FOS3组)。酶联免疫吸附试验(ELISA)测定6、12和24 h细胞培养上清液中ECM层黏连蛋白(LN),纤维连接蛋白(FN)和Ⅳ型胶原蛋白(ColⅣ)含量;荧光半定量RT-PCR检测β1整合素mRNA表达。结果体外培养的大鼠GMC可分泌一定量的ECM,在各时间点,LPS组GMC的ECM分泌均高于对照组(P<0.01),而FOS各组GMC的ECM分泌均低于LPS组(P<0.01);体外培养的大鼠GMC可表达一定量的β1整合素mRNA,在各时间点,LPS组GMC的β1整合素mRNA表达量均高于对照组,FOS各组GMC的β1整合素mRNA表达量均低于LPS组。结论 LPS可诱导大鼠GMCβ1整合素表达及ECM分泌增加,而FOS可抑制LPS诱导的大鼠GMCβ1整合素表达及ECM分泌增加,FOS可以通过抑制细胞因子及ECM分泌等非血流动力学机制对肾脏起保护作用。Objective To observe the effects of fosinopril(FOS)on β1-integrin expression and extracellular matrix(EMA)secretion in lipopolysaccharide(LPS)-induced rat glomerular mesangial cells(GMC).Methods GMC of rats were cultured in vitro.The cells at passages 3 to 10 were used in the experiment after identification.The experiment included five groups:control group,LPS group,groups with high,middle and low dose of FOS(FOS1,FOS2 and FOS3 group).The supernatant were collected 6 h,12 h,and 24 h after the cells culture.The ECM,including laminin(LN),fibronectin(FN)and type Ⅳ collagen(Col Ⅳ)was detected by the enzyme-linked immunosorbent-assay(ELISA).The expression of β1-integrin mRNA was detected by semi-quantitative real-time RT-PCR.Results Cultured GMC of rats can secrete certain amount of ECM in vitro.At each time point,the ECM secretion was significantly higher in LPS group than that in control group(P 0.01),but significantly lower in all FOS groups than that in LPS group(P 0.01).Cultured GMC of rats can express low level of β1-integrin mRNA in vitro.At each time point,β1-integrin mRNA expression was significantly higher in LPS group than that in control group,but was significantly lower in FOS groups than that in LPS group.Conclusions LPS can induce the GMC β1-integrin mRNA expression and ECM secretion in rats.Meanwhile,FOS can inhibit the GMC β1-integrin mRNA expression and ECM secretion in rats in dose-dependent manner.FOS can protect the kidneys from non-hemodynamic mechanism by inhibiting cytokine and ECM secretion
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