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
冯梓航 1刘兴龙 1赵迎春 1陈雪艳 1丁传波 1刘文丛1
作者信息
- 1. 吉林农业大学中药材学院,长春 130118
- 折叠
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1,2.5,5,10,25,50,75,100 μmol/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Hoechst 33258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.50,75,100 μmol/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75,100 μmol/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.
Abstract
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 signal-ing 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-Fructosyl-Glucose 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 inhibi-tion rate reached a plateau after 48 h.The inhibitory effect of Arginyl-Fructosyl-Glucose at 50,75 and 100 mol/L concentrations was the best,and the survival rate of B16 cells was significantly re-duced(P＜0.05,P＜0.01).RT-PCR and Western Blot assay showed that protein levels and Bcl-2 mRNA 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 in-hibit the proliferation of melanoma B16 cells and induce their apoptosis.The mechanism may be re-lated 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Key words
Arginyl-Fructosyl-Glucose/melanoma B16 cell/proliferation and apoptosis/malig-nant tumor/ginseng引用本文复制引用
基金项目
吉林省科技发展计划(20180201070YY)
出版年
2024
国家科技期刊平台
NSTL
万方数据