麝香保心丸通过 PI3K/Akt/GSK3β 通路对缺氧诱导的 HL-1 心肌细胞损伤的抗凋亡作用。
Anti-apoptotic effect of Suxiao Jiuxin Pills against hypoxia-induced injury through PI3K/Akt/GSK3β pathway in HL-1 cardiomyocytes.
机构信息
Cardiovascular Department and Cardiovascular Research Institute of Traditional Chinese Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shangha, China.
Cardiovascular Department and Cardiovascular Research Institute of Traditional Chinese Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shangha, China.
出版信息
J Chin Med Assoc. 2018 Sep;81(9):816-824. doi: 10.1016/j.jcma.2018.02.002. Epub 2018 Apr 7.
BACKGROUND
Suxiao Jiuxin Pill (SX), Chinese traditional medicine primarily consisting of tetramethylpyrazine and borneol, has been shown to protect against ischemic heart diseases. Nevertheless, the involved mechanism still remains unclear. The following study aimed to investigate the potential protective effect and molecular mechanisms of SX on apoptosis in HL-1 cardiomyocytes.
METHODS
Simulated hypoxia was established by culturing HL-1 cardiomyocytes in DMEM with no glucose or serum in a hypoxic chamber with 95% N and 5% CO for 24 h. HL-1 cardiomyocytes were divided into 5 groups: control, hypoxic injury, hypoxic injury + insulin (PI3K agonist, 10 μM), hypoxic injury + SX (100 μg/mL), and hypoxic injury + SX + LY294002 (PI3K inhibitor, 10 μM) (n = 3 wells/group). The anti-apoptotic effect of SX was evaluated by Annexin V/PI analysis. Mitochondrial membrane potential (ΔΨm) was detected by JC-1 assay. The protein expression of PI3K, phosphorylated PI3K (p-PI3K), Akt, phosphorylated Akt (p-Akt), GSK3β and phosphorylated GSK3β (p-GSK3β) were detected by western blot.
RESULTS
SX exhibited anti-apoptotic effect in HL-1 cardiomyocytes; nonetheless, the effect was blocked by PI3K inhibitor LY294002. Also, the anti-apoptotic effect of SX was mediated by increased mitochondrial membrane potential (ΔΨm). Furthermore, p-PI3K, p-Akt, and p-GSK3β expressions were significantly increased after SX treatment, while they were all reduced after administration of LY294002.
CONCLUSION
SX protects HL-1 cardiomyocytes from apoptosis induced by hypoxia, partly through enhancing the phosphorylation of PI3K/Akt/GSK3β signaling pathway.
背景
速效救心丸（SX）是一种主要由四甲基吡嗪和冰片组成的中药，已被证明可预防缺血性心脏病。然而，其涉及的机制尚不清楚。本研究旨在探讨 SX 对 HL-1 心肌细胞凋亡的潜在保护作用及其分子机制。
方法
在缺氧室中用 95% N 和 5% CO 培养无葡萄糖和血清的 DMEM 培养 HL-1 心肌细胞 24 小时，建立模拟缺氧模型。将 HL-1 心肌细胞分为 5 组：对照组、缺氧损伤组、缺氧损伤+胰岛素（PI3K 激动剂，10 μM）组、缺氧损伤+SX（100 μg/mL）组和缺氧损伤+SX+LY294002（PI3K 抑制剂，10 μM）组（n = 3 孔/组）。通过 Annexin V/PI 分析评估 SX 的抗凋亡作用。通过 JC-1 测定检测线粒体膜电位（ΔΨm）。通过 Western blot 检测 PI3K、磷酸化 PI3K（p-PI3K）、Akt、磷酸化 Akt（p-Akt）、GSK3β 和磷酸化 GSK3β（p-GSK3β）的蛋白表达。
结果
SX 对 HL-1 心肌细胞表现出抗凋亡作用；然而，该作用被 PI3K 抑制剂 LY294002 阻断。此外，SX 的抗凋亡作用是通过增加线粒体膜电位（ΔΨm）介导的。进一步的研究表明，SX 处理后 p-PI3K、p-Akt 和 p-GSK3β 的表达明显增加，而给予 LY294002 后则全部减少。
结论
SX 可保护 HL-1 心肌细胞免受缺氧诱导的凋亡，部分通过增强 PI3K/Akt/GSK3β 信号通路的磷酸化。
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