SRF-miRNA-143．KLF-4信号通路在高糖诱导的人腹膜间皮细胞表型转换中的作用

目的探讨Kruppel样因子4（KLF-4）在高糖刺激诱导的人腹膜问皮细胞（HPMC）表型转换中的作用。方法以50mmol／L葡萄糖刺激HPMC72h后，分别采用Real-timePCR及Westernblotting检测KLF-4、上皮细胞钙黏蛋白（E-cadherin）、α平滑肌肌动蛋白（α-SMA）、结缔组织生长因子（CTGF）及miRNA-143的表达。以LV-KLF-4及inhibitor转染高糖刺激72h后的HPMC，shRNA-KLF-4及mimic转染正常的HPMC，采用Real-timePCR及Westernblotting检测KLF-4、E-cadherin、α-SMA、C...

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Published inJie fang jun yi xue za zhi Vol. 41; no. 12; pp. 998 - 1004
Main Author 车明文 闫计 龚莉芳 耿秀萍 何丽洁 巨立中 王汉民
Format Journal Article
LanguageChinese
Published Beijing People's Military Medical Press 2016
Subjects
Glucose
Kruppel样因子4
人腹膜问皮细胞
细胞表型转换
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Summary:目的探讨Kruppel样因子4（KLF-4）在高糖刺激诱导的人腹膜问皮细胞（HPMC）表型转换中的作用。方法以50mmol／L葡萄糖刺激HPMC72h后，分别采用Real-timePCR及Westernblotting检测KLF-4、上皮细胞钙黏蛋白（E-cadherin）、α平滑肌肌动蛋白（α-SMA）、结缔组织生长因子（CTGF）及miRNA-143的表达。以LV-KLF-4及inhibitor转染高糖刺激72h后的HPMC，shRNA-KLF-4及mimic转染正常的HPMC，采用Real-timePCR及Westernblotting检测KLF-4、E-cadherin、α-SMA、CTGF及miRNA-143的表达。结果高糖刺激HPMC72h后，Real-timePCR检测结果显示，KLF-4表达无明显变化，E-cadherin表达下降，α-SMA、CTGF及miRNA-143表达增加。Westernblotting检测结果显示，KLF-4、E-cadherin表达下降。上调KLF-4表达后，HPMC中E-cadherin表达增加，而α-SMA、CTGF表达下降；下调KLF-4表达后，HPMC中E-cadherin表达下降，而α-SMA、CTGF表达增加（P〈0．05）。结论高糖刺激可以下调KLF-4蛋白的表达，SRF-miRNA-143-KLF-4信号通路轴参与了高糖刺激诱导的人腹膜间皮细胞表型转换过程。
Bibliography:Objective To explore the role of Kruppel-like factor 4 （KLF-4） in phenotypic transition of human peritoneal mesothelial cells （HPMCs） induced via high glucose. Methods HPMCs were induced by 50mmol/L glucose for 72 hours, the expressions of epithelium-cadherin （E-cadherin）, KLF-4, α-smooth muscle actin （α-SMA）, connective tissue growth factor （CTGF） and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. The treated cells were transfected with LV- KLF-4 and inhibitor, the untreated cells were transfected with shRNA-KLF-4 and mimic. The mRNA and protein expressions of KLF-4, E-cadherin, α-SMA, CTGF and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. Results Real-time PCR showed that the expression of E-cadherin decreased and of α-SMA, CTGF and miRNA-143 increased, but of KLF- 4 not changed in high glucose treated cells. Western blotting showed that the expression of KLF-4 and E-cadherin decreased. Upregulating KLF-4 increased the expression of E-cadherin, bu
ISSN:0577-7402
DOI:10.11855/j.issn.0577-7402.2016.12.06