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作 者:钱明[1] 甘卫华[2] 陈荣华[1] 潘晓勤[1] 费莉[1] 郭梅[1] 黄艳军[1] 徐颂周[1]
机构地区:[1]南京医科大学儿科医学研究所,江苏南京210029 [2]南京医科大学第二附属医院儿科,江苏南京210011
出 处:《南京医科大学学报(自然科学版)》2006年第2期77-81,85,共6页Journal of Nanjing Medical University(Natural Sciences)
基 金:国家自然科学基金资助项目(30100081)
摘 要:目的:探讨内源性结缔组织生长因子(CTGF)在人肾小管上皮细胞(HK2)转分化过程中的作用。方法:将HK2细胞分为4组:对照组;转化生长因子β1(TGF-β1)5ng/ml刺激组;TGF-β15ng/ml刺激+CTGF反义ODN3mmol/L干预组;TGF-β15ng/ml刺激+CTGF正义ODN3mmol/L对照组。用倒置显微镜观察各组细胞的形态学变化,用RT-PCR检测各组细胞中CTGF、上皮细胞钙粘蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、纤连蛋白(FN)mRNA表达变化。结果:TGF-β1刺激使HK2细胞形态由椭圆形转为梭形;下调E-cadherinmRNA表达,上调CTGF、α-SMA和FNmRNA表达,且CTGFmRNA表达变化在时间上早于E-cadherin、α-SMA和FNmRNA表达变化;CTGF反义ODN可对抗TGF-β1引起的细胞形态学改变和E-cadherin、CTGF、α-SMA、FNmRNA表达变化效应。结论:内源性CTGF介导了TGF-β1对HK2细胞的转分化效应。Objective: To investigate the significance of endogenous connective tissue growth factor (CTGF) in epithel ial- myofibroblast transdifferentiation of human renal tubular epithelial cell line (HK2) in vitro. Methods:Cultured HK2 were divided into four groups: ( 1 )control; (2)treated with TGF-β1 (5 ng/ml) ; (3)treated with TGF-β1 (5 ng/ml) plus CTGF antisense oligonucleotide (3 mmol/L); (4)treated with TGF-β1 (5 ng/ml) plus CTGF sense oligonucleotide (3 mmol/L). The cell morphological changes were traced with inverted microscope, and the expression of CTGF.E-cadherin.α-smooth muscle actin (α-SMA) and fibronectin(FN) mRNAs were detected by RT-PCR. Results: In HK2 cells, TGF-β1 stimulated the oval-to-fusiform transdifferentiation in morphology, downregulated E-cadherin mRNA expression, upregulated CTGF, α-SMA and FN mRNAs expression. The presence of the change of CTGF mRNA expression preceded those of E-cadherin.α-SMA and FN mRNAs; CTGF antisense oligonucleotide inhibited these effects by TGF-β1. Conclusion:Endogenous CTGF mediates the regulation of transforming growth factor β1 in human renal tubular epithelial-myofibroblast transdifferentiation in vitro.
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