鹿血晶增强巨噬细胞抗黑色素瘤作用及其机制研究

Study on anti-melanoma effect of deer blood crystal on macrophages

目的:探索鹿血晶对巨噬细胞杀伤黑色素瘤细胞的影响。方法:细胞增殖试剂盒(CCK8)检测细胞活力,流式细胞术检测细胞凋亡和巨噬细胞吞噬情况。qRT-PCR法检测炎症因子mRNA表达水平,ELISA检测上清炎症因子浓度。蛋白印迹法检测mTOR信号通路下游4EBP和S6磷酸化水平。结果:鹿血晶刺激后Raw264.7细胞培养上清抑制B16F10细胞活力,促进B16F10细胞凋亡。鹿血晶促进Raw264.7细胞吞噬B16F10细胞,增强Raw264.7细胞活力,并上调炎症因子iNOS、TNF-α、IL-6、IL-1β的mRNA表达,促进上述炎症因子分泌,上调4EBP和S6蛋白磷酸化水平。结论:鹿血晶通过活化mTOR信号通路提高巨噬细胞对黑色素瘤细胞的杀伤作用。

Objective:To explore the anti-melanoma effect of deer blood crystal(DBC)on macrophages.Methods:Cell Countion Kit-8 was used to evaluate cell viability.Flow cytometry was used to detect cell apoptosis and the phagocytic index.The mRNA expressions of inflammatory cytokines were measured by qRT-PCR,and their contents were measured by ELISA.Western blot was performed to detect the protein expression levels of 4EBP and S6,downstream of the mTOR signaling,and their phosphorylation levels.Results:Supernatant of Raw264.7 pre-treated with DBC inhibited the cell viability of B16F10,and increased the apoptosis of B16F10 as well,and promoted the phagocytosis of Raw264.7 cells to B16F10 cells.DBC significantly increased the cell viability of Raw 264.7 cells,it also elevated the mRNA expressions and secretion levels of iNOS,TNF-α,IL-6,IL-1βand up-regulated the phosphorylation levels of 4EBP and S6.Conclusion:DBC enhances the killing effect of macrophages to melanoma via regulating mTOR signaling phathway.