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  

The role of SRF- mi RNA-143-KLF-4 signal pathway in the HPMC phenotypic transition induced via high glucose

作　　者：车明文 闫计[1] 龚莉芳 耿秀萍[1] 何丽洁[2] 巨立中[1] 王汉民[2] 

机构地区：[1]解放军273医院内三科,新疆库尔勒841000 [2]第四军医大学西京医院肾脏内科,西安710032

出　　处：《解放军医学杂志》2016年第12期998-1004,共7页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81270768;81270849)~~

摘　　要：目的探讨Kruppel样因子4(KLF-4)在高糖刺激诱导的人腹膜间皮细胞(HPMC)表型转换中的作用。方法以50mmol/L葡萄糖刺激HPMC 72h后,分别采用Real-time PCR及Western blotting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mi RNA-143的表达。以LV-KLF-4及inhibitor转染高糖刺激72h后的HPMC,sh RNA-KLF-4及mimic转染正常的HPMC,采用Real-time PCR及Western blotting检测KLF-4、E-cadherin、α-SMA、CTGF及mi RNA-143的表达。结果高糖刺激HPMC 72h后,Real-time PCR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mi RNA-143表达增加。Western blotting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-mi RNA-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。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, but decreased the expression of α-SMA and CTGF. Down-regulating KLF- 4 decreased the expression of E-cadherin, but augment the expression of α-SMA and CTGF. Conclusion High glucose may induce the down-regulation of KLF-4 protein, and SRF- miRNA-143-KLF-4 signal pathway axis may be involved in the process of HPMC phenotypic transition.

关 键 词：细胞表型转换 Kruppel样因子4 人腹膜间皮细胞 

分 类 号：R459.51[医药卫生—治疗学]