α-常春藤皂苷对黑色素瘤B16细胞增殖和凋亡的影响及机制  

Effect of α-Hederin on Cell Proliferation and Apoptosis in Melanoma B16 Cells and Its Mechanism

作　　者：张步鑫[1] 赵献敏[2] 成琼 李锡清[3] ZHANG Bu-xin;ZHAO Xian-min;CHENG Qiong;LI Xi-qing(Henan Province Hospital of Traditional Chinese Medicine, Zhengzhou 450000, China;Basic Medical College, Henan University of Chinese Medicine, Zhengzhou 450000, China;Henan Provincial People＇s Hospital, Zhengzhou 450000, China)

机构地区：[1]河南省中医院,郑州450000 [2]河南中医药大学基础医学院,郑州450000 [3]河南省人民医院,郑州450000

出　　处：《中国实验方剂学杂志》2018年第12期81-85,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81673775)

摘　　要：目的：探讨α-常春藤皂苷（α-HN）对黑色素瘤B16细胞增殖和凋亡的影响及潜在机制。方法：常规培养B16细胞至对数生长期,给予不同浓度α-HN（50,100,200μmol·L^-1）处理,设未经α-HN处理的B16细胞为空白组;采用噻唑蓝（MTT）比色法检测α-HN处理24,48,72 h后各组细胞的增殖情况,采用膜联蛋白V/碘化丙啶（Annexin V-FITC/PI）凋亡试剂盒检测α-HN处理48 h后的细胞凋亡率,半胱天冬蛋白酶（Caspase）活性检测试剂盒检测Caspase-3和Caspase-9的活性,蛋白免疫印迹法（Western blot）检测B淋巴细胞瘤-2（Bcl-2）,Bcl-2相关X蛋白（Bax）,磷酸化磷脂酰肌醇-3-羟激酶（p-PI3K）,磷酸化蛋白激酶B（p-Akt）及磷酸化雷帕霉素靶蛋白（p-mTOR）的表达情况。结果：与空白组相比,经50,100,200μmol·L^-1α-HN处理24,48,72 h后,B16细胞增殖抑制率显著增加（P〈0.01）,且随着观察时间的延长,增殖抑制率有逐渐增加趋势,该抑制作用呈时间依赖性和浓度依赖性;与空白组相比,α-HN处理48 h后B16细胞的凋亡率升高,Caspase-3和Caspase-9活性增强,且Bcl-2水平降低而Bax水平升高（P〈0.01）;α-HN处理48 h后的p-PI3K,p-Akt,p-mTOR蛋白表达较空白组降低（P〈0.05,P〈0.01）。结论：α-HN具有抑制B16细胞增殖及诱导凋亡作用,其可能通过抑制PI3K/Akt/mTOR信号通路来发挥细胞毒及诱导凋亡作用,在黑色素瘤治疗中有较好的应用前景。Objective： To explore the effect of α-hederin（ α-HN） on the proliferation and apoptosis in melanoma B16 cells and its underling mechanism. Method： The melanoma B16 cells were conventionally cultivated to the logarithmic phase and treated with different concentrations of α-HN（ 50,100,200 μmol·L^-1）.B16 cells untreated with α-HN were set up as blank group. The cell proliferation was detected by 3-（ 4,5-dimethyl thiazole）-2,5-diphenyl 4 azole nitrogen bromine salt（ MTT） method after 24,48,72 h of treatment,and the apoptosis rate, activities of Caspase-3 and Caspase-9, expressions of Bcl-2, Bax, phosphorylation phosphatidyl inositol-3-hydroxy kinase（ p-PI3 K）,phosphorylation protein kinase B（ p-Akt） and phosphorylation rapamycin target protein（ p-mTOR） were evaluated by flow cytometry,Caspase activity detection kits and Western blot,respectively. Result： Compared with blank group,the cell proliferation inhibition rate in α-HN groups was increased significantly after being treated with 50,100,200 μmol·L^-1α-HN for 24,48,72 h（ P〈0. 01）,and B16 cell proliferation inhibition rate increased significantly; compared with blank group,the cell apoptosis rate was increased,the activities of Caspase-3 and Caspase-9 were enhanced,the Bcl-2 level reduced,and Bax level increased in α-HN groups,with statistically significant differences（ P〈0. 01）; compared with blank group,expressions of p-PI3 K,p-Akt and p-mTOR were reduced in α-HN groups,with statistically significant differences（ P〈0. 05,P〈0. 01）. Conclusion： α-HN has the effects in inhibiting cell proliferation and inducing apoptosis in melanoma B16 cells,which may be related with the inhibition of PI3 K/Akt/mTOR signaling pathway,with a good application prospect in the treatment of melanoma.

关 键 词：黑色素瘤 α-常春藤皂苷(α-HN) 增殖 凋亡 磷脂酰肌醇-3-羟激酶/蛋白激酶B/雷帕霉素靶蛋白(PI3K/Akt/mTOR)信号通路 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]