T3 peptide, a fragment of tumstatin, stimulates proliferation and migration of cardiac fibroblasts through activation of Akt signaling pathway
T3 peptide, a fragment of tumstatin, stimulates proliferation and migration of cardiac fibroblasts through activation of Akt signaling pathway
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DOI：
10.1007/s00210-017-1413-0
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发表时间：
2017-08
期刊：
影响因子：
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通讯作者：
Jumpei Yasuda;Kana Fukui;M. Okada;H. Yamawaki
中科院分区：
文献类型：
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作者：
Jumpei Yasuda;Kana Fukui;M. Okada;H. Yamawaki
心肌成纤维细胞的增殖和迁移在心肌梗死后创面愈合早期具有重要意义。塔司他汀是IV型胶原α3链的裂解片段，其对心脏成纤维细胞这些功能的影响尚未明确。在这项研究中，我们通过使用T3肽，一个活性片段的tumstatin检测。从成年雄性Wistar大鼠心室分离心脏成纤维细胞。细胞计数法检测细胞增殖。博伊登室法检测迁移。Western blotting检测蛋白表达和磷酸化水平。T3肽（300 ng/ml, 24 h）显著提高心肌成纤维细胞的增殖和迁移能力。T3肽（300 ng/ml, 30 min）显著提高Akt （Ser473）磷酸化水平。phosphoinositide 3-kinase (PI3K)/Akt抑制剂LY294002 （10 μM，预处理30 min）可显著抑制T3肽诱导的心肌成纤维细胞增殖、迁移和Akt信号通路的激活。整合素αvβ3/αvβ5抑制剂西伦吉肽抑制Akt磷酸化和心脏成纤维细胞增殖。大鼠心肌梗死模型梗死区抑素表达降低。我们首次证明T3肽至少部分通过结合整合素αvβ3/αvβ5激活PI3K/Akt信号通路刺激成年大鼠心脏成纤维细胞增殖和迁移。这些结果表明，塔司他汀通过激活心肌梗死后的成纤维细胞促进创面愈合的初始阶段。
Proliferation and migration of cardiac fibroblasts are important in early stage of wound-healing after myocardial infarction. The effects of tumstatin, a cleaved fragment of collagen type IV α3 chain, on these functions of cardiac fibroblasts have not been clarified. In this study, we examined it by using T3 peptide, an active fragment of tumstatin. Cardiac fibroblasts were isolated from ventricles of adult male Wistar rats. Proliferation was examined by a cell counting assay. Boyden chamber assay was performed to examine migration. Expression and phosphorylation of proteins were determined by Western blotting. T3 peptide (300 ng/ml, 24 h) significantly increased proliferation and migration of cardiac fibroblasts. T3 peptide (300 ng/ml, 30 min) significantly increased Akt (Ser473) phosphorylation. LY294002 (10 μM, 30 min pretreatment), a phosphoinositide 3-kinase (PI3K)/Akt inhibitor, significantly inhibited the T3 peptide-induced proliferation, migration, and activation of Akt signaling pathway in cardiac fibroblasts. Cilengitide, an inhibitor of integrin αvβ3/αvβ5, suppressed Akt phosphorylation and proliferation of cardiac fibroblasts. Expression of tumstatin decreased in the infarcted area of rat model of myocardial infarction. We for the first time demonstrated that T3 peptide stimulates proliferation and migration at least partly through the activation of PI3K/Akt signaling pathway via binding integrin αvβ3/αvβ5in adult rat cardiac fibroblasts. These results indicate that tumstatin promotes the initial stage of wound-healing through activation of cardiac fibroblasts after myocardial infarction.