目的 研究银杏叶提取物(GBE)调控磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路对人脑血管外膜成纤维细胞(HBVAFs)增殖和凋亡的影响.方法 将HBVAFs分为空白组(正常培养,未做处理)和低、中、高剂量实验组(25、50 和 100 mg·L-1 的 GBE 处理 24 h)及联合组(100 mg·L-1 GBE 和2 nmol·L-1胰岛素样生长因子-1处理24 h).用噻唑蓝法检测细胞的增殖情况,用流式细胞仪检测细胞的凋亡情况,用蛋白质印迹法检测细胞磷酸化PI3K(p-PI3K)、PI3K、磷酸化AKT(p-AKT)及AKT蛋白的表达水平.结果 低、中、高剂量实验组和联合组、空白组的细胞存活率分别为(66.78±8.21)％、(53.51±7.40)％、(42.33±6.01)％、(75.69±5.87)％和(96.77±12.80)％,细胞凋亡率分别为(24.83±2.47)％、(30.07±3.06)％、(37.02±4.66)％、(16.71±2.11)％和(8.99±0.19)％,p-PI3K/PI3K 比值分别为 0.75±0.07、0.62±0.06、0.43±0.04、0.72±0.05 和 0.90±0.09,p-AKT/AKT 比值分别为0.70±0.07、0.60±0.06、0.38±0.03、0.69±0.06 和 0.90±0.09.低、中、高剂量实验组的上述指标与空白组比较,联合组的上述指标与高剂量实验组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 GBE可降低HBVAFs增殖,促进凋亡,其机制与抑制PI3K/AKT信号通路相关.
Objective To study the effects of Ginkgo biloba extract(GBE)on the proliferation and apoptosis of human cerebral adventitia fibroblasts(HBVAFs)by regulating the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)signaling pathway.Methods HBVAFs cells were divided into blank group(normal culture,no treatment),experimental-L,-M,-H groups(25,50 and 100 mg·L-1 GBE treatment for 24 h)and combined group(100 mg·L-1 GBE treatment and 2 nmol·L-1 insulin-like growth factor-1 treatment for 24 h).Cell proliferation was detected by thiazole blue assay,cell apoptosis was detected by flow cytometry,and the protein expressions of phosphorylated-PI3K(p-PI3K),PI3K、phosphorylated-AKT(p-AKT)and AKT were detected by Western blot.Results The cell survival rates of experimental-L,experimental-M,experimental-H,combined and blank groups were(66.78±8.21)％,(53.51±7.40)％,(42.33±6.01)％,(75.69±5.87)％and(96.77±12.80)％;the apoptosis rates were(24.83±2.47)％,(30.07±3.06)％,(37.02±4.66)％,(16.71±2.11)％and(8.99±0.19)％;p-PI3K/PI3K ratios were 0.75±0.07,0.62±0.06,0.43±0.04,0.72±0.05 and 0.90±0.09;p-AKT/AKT ratios were 0.70±0.07,0.60±0.06,0.38±0.03,0.69±0.06 and 0.90±0.09,respectively.The above indexes of the experimental-L,-M,-H groups were compared with the blank group,and the above indexes of the combined group were compared with the experimental-H group,the differences were statistically significant(all P＜0.05).Conclusion GBE can reduce the proliferation of HBVAFs,promote apoptosis,and its mechanism is related to the inhibition of PI3K/AKT signaling pathway.
毕晓晨;孔晓彬;赵娜;顾晶晶
浙江中医药大学基础医学院实验教学中心,浙江杭州 310000浙江中医药大学基础医学院病理与病理生理教研室,浙江杭州 310000
中医学
银杏叶提取物;磷酸肌醇3激酶/蛋白激酶B信号通路;人脑血管外膜成纤维细胞;增殖;凋亡
Ginkgo biloba extract;phosphoinositol 3 kinase/protein kinase B signaling pathway;human cerebral adventitia fibroblasts;proliferation;apoptosis
《中国临床药理学杂志》 2024 (011)
1556-1559 / 4
国家自然科学基金青年科学基金资助项目(81603487)
10.13699/j.cnki.1001-6821.2024.11.003
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