To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose...To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of α-smooth muscle actin (α-SMA) were assessed by indirect immuno-fluorescence, and the percentage of α-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of α-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of α-SMA were markedly stronger than that in negative controls. The percentages of α-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9 %, 65.5 % vs 2.4 %, P<0.01) .α-SMA mRNA levels were also up-regulated by the stimulation of rhCTGF (P<0.01). These results suggest that CTGF can promote the transdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).展开更多
[目的]建立人肝星状细胞(LX-2)与人肾小管上皮细胞(HK-2)共培养体系的理论模型,研究LX-2对HK-2转分化的影响。[方法]以Transwell小室建立体外LX-2细胞和HK-2细胞间接共培养体系。应用细胞免疫组织化学法检测HK-2细胞平滑肌肌动蛋白(α-S...[目的]建立人肝星状细胞(LX-2)与人肾小管上皮细胞(HK-2)共培养体系的理论模型,研究LX-2对HK-2转分化的影响。[方法]以Transwell小室建立体外LX-2细胞和HK-2细胞间接共培养体系。应用细胞免疫组织化学法检测HK-2细胞平滑肌肌动蛋白(α-SMA)表达;用酶联免疫吸附法(ELISA法)检测各组细胞培养液中转化生长因子-β1(TGF-β1)的含量;逆转录-聚合酶链反应法(RT-PCR法)检测HK-2细胞TβRⅠm RNA、TβRⅡm RNA、Smad2 m RNA、Smad3 m RNA、Smad7 m RNA的表达。[结果]除了空白组,形态学上其他两组均可看到HK-2细胞胞浆染棕黄色,不同数量细胞由立方铺路石样转变为梭形长条状,细胞肥大;HK-2细胞高表达α-SMA(P<0.01);HK-2细胞上清液中TGF-β1含量明显高于空白组(P<0.01);HK-2细胞TβRⅠ、Smad2、Smad3基因表达明显上调,而TβRⅡ、Smad7基因表达明显下调。[结论]Transwell共培养法可诱导HK-2细胞转分化,其机制可能与TGF-β1/Smad信号通路有关。展开更多
基金ThisworkwassupportedbyagrantfromtheScience&TechnologyFoundationofHubeiProvince (No .2 0 0 3AA30 1C14 )
文摘To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of α-smooth muscle actin (α-SMA) were assessed by indirect immuno-fluorescence, and the percentage of α-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of α-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of α-SMA were markedly stronger than that in negative controls. The percentages of α-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9 %, 65.5 % vs 2.4 %, P<0.01) .α-SMA mRNA levels were also up-regulated by the stimulation of rhCTGF (P<0.01). These results suggest that CTGF can promote the transdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).
文摘[目的]建立人肝星状细胞(LX-2)与人肾小管上皮细胞(HK-2)共培养体系的理论模型,研究LX-2对HK-2转分化的影响。[方法]以Transwell小室建立体外LX-2细胞和HK-2细胞间接共培养体系。应用细胞免疫组织化学法检测HK-2细胞平滑肌肌动蛋白(α-SMA)表达;用酶联免疫吸附法(ELISA法)检测各组细胞培养液中转化生长因子-β1(TGF-β1)的含量;逆转录-聚合酶链反应法(RT-PCR法)检测HK-2细胞TβRⅠm RNA、TβRⅡm RNA、Smad2 m RNA、Smad3 m RNA、Smad7 m RNA的表达。[结果]除了空白组,形态学上其他两组均可看到HK-2细胞胞浆染棕黄色,不同数量细胞由立方铺路石样转变为梭形长条状,细胞肥大;HK-2细胞高表达α-SMA(P<0.01);HK-2细胞上清液中TGF-β1含量明显高于空白组(P<0.01);HK-2细胞TβRⅠ、Smad2、Smad3基因表达明显上调,而TβRⅡ、Smad7基因表达明显下调。[结论]Transwell共培养法可诱导HK-2细胞转分化,其机制可能与TGF-β1/Smad信号通路有关。
