Effects of bradykinin on TGF-1-induced epithelial-mesenchymal transition in ARPE-19 cells
Effects of bradykinin on TGF-1-induced epithelial-mesenchymal transition in ARPE-19 cells
复制标题
缓激肽对 TGF-1 诱导的 ARPE-19 细胞上皮间质转化的影响
DOI：
10.3892/mmr.2018.8556
复制
发表时间：
2018-04-01
影响因子：
3.4
通讯作者：
Yu, Jing
中科院分区：
文献类型：
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作者：
Wei, Qingquan;Liu, Qingyu;Yu, Jing
本研究的目的是通过暴露于转化生长因子-1 （TGF-1）来研究缓激素（BK）对视网膜色素上皮（RPE）细胞上皮-间质转化（EMT）模型的影响。目的是提高BK对增殖性玻璃体视网膜病变（PVR）进展的影响，寻找一种新的临床预防和治疗PVR的方法。倒置相差显微镜观察ARPE-19细胞形态。使用细胞计数试剂盒-8评估TGF-1对ARPE-19细胞增殖的影响。采用Western blot和免疫荧光法检测上皮标志物E-cadherin、间充质标志物平滑肌肌动蛋白（SMA）和vimentin的表达水平，以及TGF/Smad信号通路中抗十肢瘫痪同源物（Smad）3和Smad7的磷酸化(p)母细胞的表达水平。创面愈合试验和Transwell试验检测细胞迁移能力。采用逆转录-定量聚合酶链反应（RT-qPCR）方法检测pSmad3和Smad7在TGF/Smad信号通路中的表达水平。结果显示，添加10 ng/ml TGF-1可导致ARPE-19细胞中EMT相关因子的表达。BK降低了间充质标志物-SMA和vimentin的表达水平，增加了上皮标志物E-cadherin的表达。BK在tgf -1诱导的EMT中减少细胞迁移。这些作用被一种特异性bk2受体拮抗剂HOE-140逆转。在tgf -1处理的ARPE-19细胞中，BK显著下调pSmad3的表达，上调Smad7的表达，而BK产生的保护性改变被HOE-140抑制。综上所述，10 ng/ml TGF-1可导致ARPE-19细胞发生EMT，而BK在TGF-1诱导的EMT中起负向作用。BK通过上调TGF-/Smad信号通路中Smad7的表达，下调pSmad3的表达，在TGF-1诱导的EMT中发挥作用，提示BK可能是一种新的有效治疗PVR的方法。
The aim of the present study was to investigate the effects of bradykinin (BK) on an epithelial-mesenchymal transition (EMT) model in retinal pigment epithelium (RPE) cells through exposure to transforming growth factor-1 (TGF-1). The aim was to improve the effect of BK on proliferative vitreoretinopathy (PVR) progression, and to find a novel method of clinical prevention and treatment for PVR. The morphology of ARPE-19 cells was observed using an inverted phase-contrast microscope. A Cell Counting Kit-8 was used to assess the effects of TGF-1 on the proliferation of ARPE-19 cells. Western blotting and immunofluorescence were used to detect the expression levels of the epithelial marker E-cadherin, mesenchymal markers -smooth muscle actin (SMA) and vimentin, and phosphorylated (p) mothers against decapentaplegic homolog (Smad)3 and Smad7 of the TGF/Smad signaling pathway. Wound healing tests and Transwell assays were performed to detect cell migration ability. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis was performed to detect the expression levels of pSmad3 and Smad7 in the TGF/Smad signaling pathway. The results revealed that the addition of 10 ng/ml TGF-1 resulted in the expression of factors associated with EMT in ARPE-19 cells. BK decreased the expression levels of the mesenchymal markers -SMA and vimentin, and increased the expression of the epithelial marker E-cadherin. BK decreased cell migration in TGF-1-induced EMT. These effects were reversed by HOE-140, a specific BK 2 receptor antagonist. BK significantly downregulated the expression of pSmad3 and upregulated the expression of Smad7 in TGF-1-treated ARPE-19 cells, and the protective alterations produced by BK were inhibited by HOE-140. In conclusion, 10 ng/ml TGF-1 resulted in EMT in ARPE-19 cells and BK served a negative role in TGF-1-induced EMT. BK had effects in TGF-1-induced EMT by upregulating the expression of Smad7 and downregulating the expression of pSmad3 in TGF-/Smad signaling pathway, indicating that BK may be a novel and effective therapy for PVR.