蒙花苷通过PI3K-Akt/m-TOR信号通路保护H9c2心肌细胞缺氧复氧损伤

Protective Effect of Linarin on Hypoxia-reoxygenation Injury of H9c2 Cardiomyocytes by PI3K-Akt/m-TOR Signaling Pathway

目的:探索蒙花苷对H9c2心肌细胞缺氧复氧损伤的保护作用机制,并探寻抑制心肌细胞凋亡可能的信号通路。方法:H9c2心肌细胞分为正常细胞对照组、缺氧复氧损伤模型组、不同浓度蒙花苷(12.50、25.00、50.00、100.00、200.00μg/mL)组、LY294002(50.00μg/mL)组、蒙花苷(50.00μg/mL)+LY294002(50.00μg/mL)组,共9组。检测相应组别细胞增殖活力和心肌细胞中MDA、SOD、LDH水平;采用Western blot检测PI3K/Akt/mTOR通路中关键蛋白的表达情况。结果:细胞增殖活力检测结果显示蒙花苷对细胞无毒性;H9c2心肌细胞缺氧复氧各4 h后,与正常对照组比较,模型组LDH、MDA水平显著升高,SOD水平显著降低(P<0.05)。与模型组比较,蒙花苷组p-PI3K表达及Akt磷酸化程度及凋亡蛋白Bcl-2表达显著升高,Bax表达显著降低(P<0.05)。蒙花苷对缺氧复氧损伤H9c2细胞的保护作用可被抑制剂LY294002抑制。结论:蒙花苷可通过PI3K/Akt/m-TOR信号通路抑制H9c2细胞凋亡。

Objective: To explore the protective mechanism of linarin on hypoxia-reoxygenation(HR)injury of H9 c2 cardiomyocytes, and to explore the possible signaling pathways that inhibited the apoptosis of cardiomyocytes.Methods: H9 c2 cardiomyocytes were divided into nine groups, including normal control group, hypoxia-reoxygenation injury model group, different concentrations of linarin groups(12.50,25.00,50.00,100.00,200.00 μg/mL),LY294002(50.00 μg/mL)group, 50.00 μg/mL of linarin with 50.00 μg/mL LY294002 group.Cell proliferation activity and levels of MDA,SOD and LDH in cardiomyocytes of corresponding group were detected.Western blot was used to detect the expressions of key proteins in the PI3 K/Akt/mTOR pathway.Results: The results of cell proliferation activity test showed that linarin was not toxic to cells;After hypoxia-reoxygenation injury of H9 c2 cardiomyocytes for 4 hours, compared with the normal control group, the levels of LDH and MDA in the model group were significantly increased, and the level of SOD was significantly reduced(P<0.05).Compared with the model group, the expression of p-PI3 K,the phosphorylation level of Akt and the expression of apoptotic protein Bcl-2 were significantly increased and the expression of Bax was significantly decreased in the linarin group(P<0.05).After the addition of inhibitor LY294002,the protective effect of linarin on hypoxia-reoxygenation injury of H9 c2 cells was inhibited.Conclusion: Linarin can inhibit the apoptosis of H9 c2 cells through the PI3 K/Akt/m-TOR signaling pathway.