松弛素对TGF-β诱导的内皮细胞间质化的抑制作用

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Abstract Objective: To investgate the effect of relaxin which inducing endothelial to mesenchymal transition in vitro. Methods: Endothelial cells were isolated from human umbilical vein endothelial cells (HUVECs). TGF-β10 ng/mL was added to medium for inducing endothelial to mesenchymal transition. Pretreatment relaxin 100 ng/mL and 200 ng/mL was added to medium, then co-cultured with TGF-β10 ng/mL for 48 h. The changes of morphology were observed under an inverted phase contrast microscope. CCK-8 assay was used to evaluate the cell proliferation as well as Transwell tested cell migration. The expression of CD31, vimentin, ve-cadherin andα-SMA were determined by immunofluoresence staining and western blot analysis. The co-expression of CD31 with vimentin and VWF withα-SMA were also determined by immunofluoresence. Results: A paving stone-like growth was observed in control group while TGF-βinduced more spreading and migrating cells which was called EMT. However, this transition could be prevented by relaxin. Compared with the control group, the proliferation and the mobility were increased by TGF-β(P<0.05), but declined when combining with relaxin compared to adding TGF-βonly (P<0.05). Compared with the control group, the specific protein of endothelial CD31 (0.36adding TGF-β, but the specific protein of mesenchymal vimentin (0.72±0.102 VS 0.21±0.081) and α-SMA(0.88±0.084 VS 0.24±0.046) were significantly increased. However, relaxin could inhibit the phenotype switch. Compared with the TGF-βinducing group, CD31 (0.67±0.09, 0.59±0.12 VS 0.36±0.076) and ve-cadherin(0.85±0.09, 0.72±0.064 VS 0.54±0.046) were significantly increased, but vimentin (0.56±0.011, 0.48±0.06 VS 0.72±0.102) andα-SMA (0.65±0.081, 0.54±0.032 VS 0.88±0.084) were significantly decreased when combining with relaxin. Besides, it exhibited dose-dependent. Last but not least, the number of double positive cells was largely increased in TGF-βinducing group compared to the control group. And compared with the TGF-β group, double positive cells performing mesenchymal are declined while performing endothelial ones are increased in the combined group. Conclusion: Relaxin exhibits the inhibition effect for TGF-βinduced endothelial to mesenchymal transition in vitro.

Key words： relaxin endothelial to mesenchymal transition TGF-β myocardial fibrosis

Received: 28 November 2013

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	CHEN Xiao1
	ZHOU Hao1
	ZHOU Xi1
	CAI Jiejie1
	CHEN Lingzhi2
	ZHENG Gaoshu1
	HUANG Weijian1
	ZHANG huaiqin1.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2014/V44/I3/161

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