葡萄籽原花青素通过PI3K/Akt信号通路抑制氧化应激诱导的H9c2心肌细胞凋亡

Grape Seed Proanthocyanidins Protect H9c2 Cardiomyocytes from Hydrogen Peroxide-induced Damage via PI3K/Akt Signaling Pathway

为了探讨葡萄籽原花青素(grape seed proanthocyanidin,GSP)对心肌细胞的保护作用及机制,通过CCK-8法评估细胞活力,采用Western-blot分析评估GSP对凋亡相关蛋白质(cleaved caspase-3、Bax和Bcl-2)和PI3K/Akt通路相关蛋白质(p-PI3K、PI3K、p-Akt和Akt)表达水平的影响,并使用TUNEL染色和Hoechst 33258染色评估H9c2心肌细胞凋亡情况。结果显示,GSP可以抑制H_(2)O_(2)诱导的H9c2心肌细胞的细胞毒性和凋亡,使促凋亡蛋白cleaved caspase-3和Bax表达下降,并使抗凋亡蛋白Bcl-2表达水平升高;GSP作用于H9c2细胞后,PI3K和Akt的磷酸化水平增加,使PI3K/Akt信号通路激活。实验结果初步表明,GSP可抑制氧化应激诱导的H9c2心肌细胞凋亡,其作用机制可能与激活PI3K/Akt信号通路有关。

To investigate protective effects and mechanism of grape seed proanthocyanidin(GSP)on H9c2 cardiomyocyte,CCK-8 assay was used to evaluate cell viability,Western-blot was used to evaluate the effects of GSP on apoptosis-related proteins(cleaved caspase-3,Bax and Bcl-2)and PI3K/Akt pathway-related proteins(p-PI3K,PI3K,p-Akt and Akt),and TUNEL and Hoechst 33258 stainings were used to evaluate the apoptosis of H9c2 cardiomyocytes.The results showed that GSP can prevent the cytotoxicity and apoptosis of H9c2 cardiomyocytes induced by H_(2)O_(2).GSP decreased the expression of pro-apoptotic proteins cleaved caspase-3 and Bax,and increased the expression of anti-apoptotic protein Bcl-2.When GSP acted on H9c2 cells,the phosphorylation levels of PI3K and Akt increased,leading to activation of the PI3K/Akt signaling pathway.The experiments preliminarily demonstrated that GSP can inhibit H9c2 cardiomyocyte apoptosis induced by oxidative stress,and its mechanism of action may be related to PI3K/Akt signaling pathway activation.