高盐对血管内皮生长因子受体-3^+巨噬细胞表型及功能的影响  

Effects of high salt concentration on the phenotypes and functions of VEGFR-3^+ macrophages

作　　者：余芳芳 杨国红[2] 李铭 陶燕燕 王秀娟 牛秀珑[2] 李玉明[2] 赵季红[2] YU Fang-fang;YANG Guo-hong;LI Ming;TAO Yan-yan;WANG Xiu-juan;NIU Xiu-long;LI Yu-ming;ZHAO Ji-hong(Graduate School,Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China;Institute of Cardiovascular Disease and Heart Center,Pingjin Hospital,Logistics University of the Chinese People's Armed Police Forces,Tianjin Key Laboratory of Cardiovascular Remodeling and Target Organ Injury)

机构地区：[1]天津中医药大学研究生院,300193 [2]武警后勤学院附属医院心脏中心、心血管病研究所、天津市心血管重塑与靶器官损伤重点实验室

出　　处：《天津医药》2018年第12期1257-1262,1369,共7页Tianjin Medical Journal

基　　金：国家自然科学基金项目(81600328);天津市自然科学基金项目(16JCQNJC11800);武警后勤学院中心实验室开放基金项目(2015ZXKF11).

摘　　要：目的观察高盐对血管内皮生长因子受体(VEGFR)-3^+巨噬细胞表型、淋巴管内皮细胞特性及功能的影响。方法利用流式分选将小鼠RAW264.7巨噬细胞中VEGFR-3^+亚群分选出来,分为Control组、低盐组(LS组,20mmol/L NaCl)和高盐组(HS组,40 mmol/L NaCl)。利用CCK-8法观察不同组VEGFR-3^+细胞活性;Real-time PCR检测NaCl干预后VEGFR-3^+巨噬细胞表型变化及淋巴管内皮细胞标志物mRNA表达水平;Transwell实验检测各组细胞的迁移功能,流式细胞术检测不同组细胞的吞噬能力。结果与Control组相比,高盐干预可以使VEGFR-3^+巨噬细胞的白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α、CC类趋化因子配体(CCL)2、血管内皮生长因子(VEGF)-C及张力应答性增强子结合蛋白(TonEBP)mRNA表达水平上调(P<0.05);HS组在高盐干预24、48 h后细胞活性均显著低于LS组(P<0.05);同时,HS组细胞迁移能力及细胞的吞噬能力与Control组相比显著增强,差异均有统计学意义(P<0.05)。结论高盐可使VEGFR-3^+巨噬细胞向M1型巨噬细胞偏移并表现出促淋巴管生成的特性,其迁移及吞噬能力显著增强,为进一步研究该亚群与淋巴管生成及心血管疾病的关系提供了依据。Objective To investigate the effect of high salt concentration on the phenotypes,lymphatic endothelial cell characteristics and functions of endothelial growth factor receptor(VEGFR)-3^+macrophages.Methods The VEGFR-3^+subsets in mouse RAW264.7macrophages were sorted by flow cytometry and divided into control group,low salt group(LS group,20mmol/L NaCl)and high salt group(HS group,40mmol/L NaCl).The viability of VEGFR-3^+cells was measured by CCK-8method in different groups.Real-time PCR was used to detect the changes of VEGFR-3+macrophage phenotypes and the mRNA expression levels of lymphatic endothelial cell markers after the intervention.Transwell assay was used to detect the migration of three groups.The phagocytosis ability was measured by flow cytometry.Results The levels of IL-1β,TNF-α,CCL2,VEGF-C and TonEBP mRNA were significantly increased in HS group compared with those of control group(P<0.05).After24h and48h high salt intervention,the viability of VEGFR-3^+macrophages was significantly decreased in HS group compared with those of LS group(P<0.05).Meanwhile,the migration and the phagocytosis ability were also significantly increased in HS group compared with those of control group(P<0.05).Conclusion The VEGFR-3^+macrophages can migrate to M1-type macrophages and show lymphangiogenesis-promoting characteristics after high salt intervention,which provides a theoretical basis for our further study about the relationships between this subpopulation and lymphangiogenesis and cardiovascular diseases.

关 键 词：巨噬细胞 免疫表型分型 淋巴管生成 血管内皮生长因子受体3 高盐 

分 类 号：R348.5[医药卫生—基础医学]