a Department of Cardiology, Affiliated Hospital of Jining Medical University , Jining , China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):2980-2988. doi: 10.1080/21691401.2019.1640708.
Myocardial ischemia is a troublesome disease. Bilobalide possesses multiple biological functions. We researched the consequents of bilobalide in OGD-irritated H9c2 cells. OGD-stimulated H9c2 cells were treated by bilobalide, and/or transfected with miR-27a inhibitor or negative control. Use CCK-8 and flow cytometry to test cell activity and apoptosis, respectively. Luciferase activity experiment was to test targeting link between miR-27a and Tmub1. Levels of cell-cycle and apoptosis relative proteins and phosphorylation of PI3K/AKT and Wnt/β-catenin related proteins were detected through western blot. OGD stimulation reduced cell activity and negatively regulated the expression of CDK4, CDK6 and CyclinD1. Cell apoptosis was increased and its related proteins were affected by OGD. Bilobalide administration reversed all the results above caused by OGD. OGD negatively regulated miR-27a while bilobalide upregulated miR-27a. miR-27a's target gene was Tmub1. The protection consequents of bilobalide were suppressed when cells were transfected with a miR-27a inhibitor that cell activity was reduced and apoptosis was raised. Attenuation in the phosphorylation level of PI3K, AKT and β-catenin by OGD was reversed by bilobalide, whereas there were opposite results after transfected with miR-27a inhibitor. Bilobalide relieved OGD-caused H9c2 cell damage, raising cell activity and attenuating apoptosis via upregulating miR-27a and activating of PI3K/AKT and Wnt/β-catenin signal pathway. Highlights Bilobalide alleviates OGD-induced H9c2 cell injury. Bilobalide upregulates miR-27a expression in OGD-stimulated H9c2 cells. Bilobalide alleviates cell injury by upregulation of miR-27a. Bilobalide actuates PI3K/AKT and Wnt/β-catenin pathways.
心肌缺血是一种棘手的疾病。白果内酯具有多种生物学功能。我们研究了白果内酯在 OGD 刺激的 H9c2 细胞中的后果。用 OGD 刺激 H9c2 细胞，并用白果内酯处理，和/或转染 miR-27a 抑制剂或阴性对照。分别用 CCK-8 和流式细胞术检测细胞活力和细胞凋亡。荧光素酶活性实验是为了检测 miR-27a 和 Tmub1 之间的靶向关系。通过 Western blot 检测细胞周期和凋亡相关蛋白以及 PI3K/AKT 和 Wnt/β-catenin 相关蛋白的磷酸化水平。OGD 刺激降低细胞活力，并负调控 CDK4、CDK6 和 CyclinD1 的表达。OGD 还增加了细胞凋亡，并影响其相关蛋白。白果内酯给药逆转了 OGD 引起的所有上述结果。OGD 负调控 miR-27a，而白果内酯上调 miR-27a。miR-27a 的靶基因是 Tmub1。当细胞转染 miR-27a 抑制剂时，白果内酯的保护作用受到抑制，细胞活力降低，凋亡增加。OGD 引起的 PI3K、AKT 和 β-catenin 磷酸化水平的衰减被白果内酯逆转，而转染 miR-27a 抑制剂后则出现相反的结果。白果内酯通过上调 miR-27a 并激活 PI3K/AKT 和 Wnt/β-catenin 信号通路，缓解 OGD 引起的 H9c2 细胞损伤，提高细胞活力，减轻细胞凋亡。重点白果内酯减轻 OGD 引起的 H9c2 细胞损伤。白果内酯上调 OGD 刺激的 H9c2 细胞中 miR-27a 的表达。白果内酯通过上调 miR-27a 减轻细胞损伤。白果内酯激活 PI3K/AKT 和 Wnt/β-catenin 通路。
