Secreted frizzled related protein 1 protects H9C2 cells from hypoxia/re-oxygenation injury by blocking the Wnt signaling pathway.
Secreted frizzled related protein 1 protects H9C2 cells from hypoxia/re-oxygenation injury by blocking the Wnt signaling pathway.
复制标题
分泌的卷曲相关蛋白1通过阻断Wnt信号通路保护H9C2细胞免受缺氧/复氧损伤
DOI：
10.1186/s12944-016-0240-5
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发表时间：
2016-04-06
影响因子：
4.5
通讯作者：
Li HY
中科院分区：
文献类型：
--
作者：
Tao J;Abudoukelimu M;Ma YT;Yang YN;Li XM;Chen BD;Liu F;He CH;Li HY
在动物模型中，抑制Wnt信号通路的分泌卷曲相关蛋白1 （strp1）是有益的，因为strp1可以减少心肌凋亡并防止心力衰竭。介导srp1细胞存活作用的机制尚未完全阐明。本研究旨在通过体外缺血/再灌注模型研究strp1对心肌细胞可能的保护作用，并评估Wnt信号通路可能参与其中。方法采用重组AAV9载体将strp1基因转染H9C2大鼠成心肌细胞，建立体外缺血再灌注模型。采用CKK-8和台盼蓝排除法测定细胞活力。Western blot检测Dvl-1、β-catenin、c-Myc、Bax、Bcl-2的表达。流式细胞术检测心肌细胞凋亡。结果我们证实strp1显著提高细胞活力（通过台盼蓝和CKK-8检测），减少细胞凋亡（通过流式细胞术分析和Bax/Bcl-2比值检测）。这些作用部分归因于strp1下调Wnt信号通路的能力（通过Western blot检测Dvl-1、β-catenin和c-Myc的表达）。事实上，用特定的激活剂Licl重新激活Wnt信号通路活性，可以降低srp1在缺氧和再氧化期间诱导的心脏保护作用。结论本研究表明，Sfrp1通过抑制Wnt信号通路，直接保护H9C2细胞免受缺氧和再氧诱导的再灌注损伤和凋亡，为Sfrp1对缺血损伤的心脏保护作用提供了新的机制认识。
BackgroundIn animal models, secreted frizzled related protein 1 (Sfrp1) inhibition of the Wnt signaling pathway is beneficial because Sfrp1 reduces myocardial apoptosis and prevents heart failure. The mechanisms mediating the cellular survival effect of Sfrp1 has not been completely elucidated. The present study was designed to investigate the possible protective actions of Sfrp1 on cardiac muscle cells using an in vitro model of ischemia/reperfusion, and to evaluate the possible involvement of the Wnt signaling pathway.MethodsWe used a recombinant AAV9 vector to deliver the Sfrp1 gene into H9C2 rat cardiomyoblasts and adopted an in vitro model of ischemia/reperfusion. Cell vitality was measured by CKK-8 and the trypan blue exclusion assay. Western blot was used to evaluate the expression of Dvl-1, β-catenin, c-Myc, Bax, and Bcl-2. Flow cytometry analysis of cardiomyocyte apoptosis was performed.ResultsWe confirmed that Sfrp1 significantly increased cell viability (assayed by trypan blue and CKK-8) and decreased apoptosis (assayed by flow cytometry analysis and the Bax/Bcl-2 ratio). These effects were partly attributable to the ability of Sfrp1 to down-regulate Wnt signaling pathway (assayed by Western blot to evaluate the expression of Dvl-1, β-catenin, and c-Myc). Indeed, reactivation of the Wnt signaling pathway activity with the specific activator, Licl, reduced Sfrp1-induced cardioprotection during hypoxia and reoxygenation.ConclusionsThe present study demonstrated that Sfrp1 directly protected H9C2 cells from hypoxia and reoxygenation-induced reperfusion injury and apoptosis through inhibition of the Wnt signaling pathway, and added new mechanistic insight regarding the cardioprotective role of Sfrp1 on ischemic damage.