精氨酸双糖苷对黑色素瘤B16细胞的作用及其机制初步研究  

Effect of Arginyl-Fructosyl-Glucose on Melanoma B16 Cells and Its Mechanism

作　　者：冯梓航 刘兴龙[1] 赵迎春 陈雪艳 丁传波[1] 刘文丛[1] FENG Zihang;LIU Xinglong;ZHAO Yingchun;CHEN Xueyan;DING Chuanbo;LIU Wencong(College of Chinese Medicinal Materials,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学中药材学院,长春130118

出　　处：《吉林农业大学学报》2024年第2期272-279,共8页Journal of Jilin Agricultural University

基　　金：吉林省科技发展计划项目(20180201070YY)。

摘　　要：研究精氨酸双糖苷(Arginyl-Fructosyl-Glucose,AFG)抑制黑色素瘤B16细胞的增殖及诱导凋亡作用,并初步探讨了其Bcl-2、Caspase-3、Bax信号通路的作用机制。培养黑色素瘤B16细胞,配制1,2.5,5,10,25,50,75,100μmol/L的精氨酸双糖苷作用于黑色素瘤B16细胞,48 h后利用CKK-8法检测精氨酸双糖苷对B16细胞增殖影响;采用LDH检测给药后细胞乳酸脱氢酶(Lactate dehydrogenase,LDH)表达水平;用Hoechst33258荧光染色法观察精氨酸双糖苷对B16细胞凋亡形态的影响;采用RT-PCR和Western Blot法分析精氨酸双糖苷对B16细胞Bcl-2、Caspase-3、Bax mRNA和蛋白表达的影响。结果表明:随着给药浓度的增大和培养时间的增加,精氨酸双糖苷对B16细胞的抑制率增加,48 h后抑制率达到平台期。50,75,100μmol/L浓度的精氨酸双糖苷抑制效果最佳,B16细胞存活率显著降低(P<0.05,P<0.01),可明显观察到B16细胞凋亡;RT-PCR和Western Blot试验发现,精氨酸双糖苷75,100μmol/L组B16细胞Bcl-2 mRNA和蛋白水平明显减少(P<0.01),Bax和Caspase-3 mRNA和蛋白水平显著升高(P<0.01)。精氨酸双糖苷可抑制黑色素瘤B16细胞增殖,诱导其凋亡,其机制可能与上调Bcl-2、Caspase-3,下调Bax有关。To observe the inhibition of Arginyl-Fructosyl-Glucose(AFG)on the proliferation and apoptosis of melanoma B16 cells,and discuss the mechanism of its Bcl-2,Caspase-3 and Bax signaling pathways preliminarily,melanoma B16 cells were cultured.1,2.5,5,10,25,50,75 and 100 mol/L of Arginyl-Fructosyl-Glucose were prepared to act on melanoma B16 cells.CKK-8 method was used to detect the effect of Arginyl-Fructosyl-Glucose on the proliferation of B16 cells after 48h;The expression level of LDH was detected by LDH after treatment;The effect of Arginyl-FructosylGlucose on apoptosis of B16 cells was observed by Hoechst 33258 fluorescence staining;RT-PCR and Western Blot were used to analyze the effect of Arginyl-Fructosyl-Glucose on the expression of Bcl-2,Caspase-3,Bax mRNA and protein in B16 cells.With the increase of drug concentration and culture time,the inhibition rate of Arginyl-Fructosyl-Glucose on B16 cells increased,and the inhibition rate reached a plateau after 48 h.The inhibitory effect of Arginyl-Fructosyl-Glucose at 50,75and 100 mol/L concentrations was the best,and the survival rate of B16 cells was significantly reduced(P<0.05,P<0.01).RT-PCR and Western Blot assay showed that protein levels and Bcl-2mRNA in B16 cells in the 75 and 100 mol/L groups significantly decreased(P<0.01),and Caspase-3 mRNA and protein levels significantly increased(P<0.01).Arginyl-Fructosyl-Glucose can inhibit the proliferation of melanoma B16 cells and induce their apoptosis.The mechanism may be related to the upregulation of Bcl-2,Caspase-3 and the downregulation of Bax.

关 键 词：精氨酸双糖苷 黑色素瘤B16细胞 增殖凋亡 恶性肿瘤 人参 

分 类 号：R285[医药卫生—中药学]