基于PI3K/Akt信号通路探讨葡萄籽原花青素对人皮肤鳞状细胞癌A431细胞凋亡的作用研究  

Grape seedproanthocyanidins induces A431 cell apoptosis through PI3K/Akt pathway

作　　者：李晓民 梁韦巍 韩志强 李君 范崇帅 刘志香 LI Xiaomin;LIANG Weiwei;HAN Zhiqiang;LI Jun;FAN Chongshuai;LIU Zhixiang(Weifang Institute of Dermatology,Weifang 261041,China)

机构地区：[1]潍坊市皮肤病防治所,261041

出　　处：《环球中医药》2022年第5期759-763,共5页Global Traditional Chinese Medicine

基　　金：潍坊市科技发展计划(2020YX078);潍坊市卫健委科研项目计划(WFWSJK-2020-129)。

摘　　要：目的基于PI3K/Akt信号通路探讨葡萄籽原花青素对人皮肤鳞状细胞癌A431细胞凋亡的影响。方法体外培养人皮肤鳞状细胞癌A431细胞,用不同浓度的葡萄籽原花青素(5、10、20、40、80μg/mL)处理24、48、72小时,MTT法检测各组细胞的活力;蛋白免疫印迹法(western blot,WB)检测cleaved caspase-3蛋白的表达及PI3K/Akt磷酸化变化。使用PI3K激活剂740-Y-P或Akt激活剂SC79处理后,观察对细胞存活率和PI3K/Akt蛋白磷酸化的影响。结果与正常对照组比较,葡萄籽原花青素组(5、10、20、40、80μg/mL)处理24、48、72小时后细胞活力显著下降(P<0.01),呈浓度、时间依赖性。Western blot结果显示,与正常对照组比较,葡萄籽原花青素组(40、80μg/mL)cleaved caspase-3蛋白表达显著增加(P<0.01),PI3K/Akt通路蛋白磷酸化水平显著降低(P<0.01)。与葡萄籽原花青素组比较,740-Y-P联合用药组、SC79联合用药组PI3K/Akt通路蛋白磷酸化水平显著升高(P<0.05),细胞相对存活率显著增加(P<0.01)。结论葡萄籽原花青素可能通过抑制PI3K/Akt信号通路诱导人皮肤鳞状细胞癌A431细胞凋亡。Objective To investigate the effect of grape seed proanthocyanidins(GSP)on apoptosis of human skin squamous cell carcinoma A431 cells based on PI3K/Akt signal pathway.Methods Human skin squamous cell carcinoma A431 cells was cultured in vitro.24,48,72 hours after different concentrations of GSP(5,10,20,40,80μg/mL)was administrated,cell viability in different groups was detected by MTT detection assay.The expression of cleaved caspase-3 protein and PI3K/Akt phosphorylation were detected by Western blot.After treated with PI3K activator 740-Y-P or Akt activator SC79,cell viability was detected by MTT and PI3K/Akt phosphorylation were determined.Results Compared with control group,cell viability was significantly decreased after treated with GSP(5,10,20,40,80μg/mL)for 24,48 and 72 hours.GSP produced significant cytotoxicity to A431 cells in a concentration dependent manner.Western blot results showed that GSP with concentrations of 40 and 80μg/mL could significantly up-regulate the expression of cleaved caspase-3 protein and down-regulate the phosphorylation of PI3K/Akt pathway protein(P<0.01).740-Y-P or SC79 combined with GSP could enhance the protein expression of phosphorylated PI3K/Akt(P<0.05).The cell viability was also significantly increased after combination treatment(P<0.01).Conclusion GSP may reduce the activity of human skin squamous cell carcinoma A431 cells and induce apoptosis by inhibiting PI3K/Akt pathway.

关 键 词：葡萄籽原花青素 鳞状细胞癌 凋亡 PI3K/AKT 通路 A431细胞 

分 类 号：R285.5[医药卫生—中药学]