二甲双胍脂质体靶向增强TAMs糖摄取对B16-TAMs共培养体系糖分布的影响  

Metformin Liposome Specifically Enhanced Glucose Uptake of TAMs for the Alteration of Glucose Distribution in B16-TAMs Co-Culture System

作　　者：郭婉 秦文涛 贝云成 张冬梅[1] GUO Wan;QIN Wentao;BEI Yuncheng;ZHANG Dongmei(College of Life Sciences,Nanjing University,Nanjing 210023,China;State Key Laboratory of Natural Medicines,China Pharmaceutical University,Nanjing 210009,China)

机构地区：[1]南京大学生命科学学院,南京210023 [2]中国药科大学天然药物活性组分与药效国家重点实验室,南京210009

出　　处：《中国细胞生物学学报》2022年第5期836-847,共12页Chinese Journal of Cell Biology

基　　金：江苏省重点研发计划(社会发展项目)(批准号:BE2020687);中国药科大学天然药物活性组分与药效国家重点实验室开放课题(批准号:SKLNMKF202109)资助的课题。

摘　　要：该文探讨了二甲双胍脂质体(metformin liposome,Met-lipo)通过靶向增强肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)的葡萄糖摄取以改变小鼠黑色素瘤细胞株B16与TAMs共培养体系中葡萄糖分布,以及抑制B16细胞增殖与迁移的可能性。利用2-NBDG作为葡萄糖类似物模拟细胞对葡萄糖的摄取;接着通过2-NBDG摄取实验、葡萄糖/乳酸试剂盒检测二甲双胍(metformin,Met)对TAMs葡萄糖摄取的影响,以及经Met预处理的TAMs改变B16-TAMs共培养体系中葡萄糖分布的能力。用细胞划痕实验、细胞增殖实验、q-PCR实验评价Met预处理的TAMs抑制B16细胞增殖与迁移的能力。同时,通过硫酸铵梯度法制备了Met-lipo,并对其粒径、电势、稳定性、包封率进行表征。通过2-NBDG摄取实验、细胞划痕实验、细胞增殖实验、q-PCR实验检测Met-lipo改变B16-TAMs接触共培养体系中葡萄糖分布的能力和其抗肿瘤效果。结果显示,Met(10 mmol/L)的处理能够有效增强TAMs的糖摄取效率,并实现对B16细胞的葡萄糖剥夺。而制备得到的Met-lipo能特异性地增强B16-TAMs共培养体系中TAMs的糖摄取能力,并展示出远强于游离Met的抗肿瘤效应。综上,Met-lipo能够靶向增强TAMs葡萄糖摄取从而改变B16-TAMs共培养体系的葡萄糖分布,导致B16增殖与迁移能力的下降。This study investigated the possibility that Met-lipo (metformin liposome) altered glucose distribution within B16-TAMs co-culture system and inhibited the growth of B16 cells by specifically enhancing glucose uptake of TAMs (tumor-associated macrophages).2-NBDG was used as glucose analogue to simulate glucose uptake of cells.The effect of Met (metformin) on glucose uptake of TAMs was detected by 2-NBDG uptake assay and glucose/lactate kits.The ability of Met-treated TAMs to alter glucose distribution within B16-TAMs co-culture system was also evaluated.Wound healing assay,cell proliferation assay and q-PCR were used to assess the ability of Met-treated TAMs to inhibit the proliferation and migration of B16 cells.Meanwhile,Met-lipo was prepared by ammonium sulfate gradient method and particle size,zeta potential,stability and drug-loading efficiency of liposome were detected by DLS and HPLC.Furthermore,the antitumor effect and ability of Met-lipo to alter glucose distribution within B16-TAMs contact co-culture system was also evaluated by 2-NBDG uptake assay,wound healing assay,cell proliferation assay and q-PCR analysis.Results showed that the treatment of Met (10 mmol/L) could effectively enhance the glucose uptake efficiency of TAMs,which could deprive the glucose B16 cells needed.Notably,the prepared Met-lipo could specifically enhance the glucose uptake of TAMs in B16-TAMs co-culture system and exhibited more prominent antitumor effect than free Met.In conclusion,Met-lipo could specifically enhance glucose uptake of TAMs and alter the glucose distribution within B16-TAMs co-culture system,and then inhibit the proliferation and migration of B16 cells.

关 键 词：肿瘤相关巨噬细胞 二甲双胍 糖摄取 脂质体 

分 类 号：R965[医药卫生—药理学]