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

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。

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

Objective To investigate the effect of high salt concentration on the phenotypes, lymphatic endothelial cellcharacteristics and functions of endothelial growth factor receptor (VEGFR)-3+ macrophages. Methods The VEGFR-3+subsets in mouse RAW264.7 macrophages were sorted by flow cytometry and divided into control group, low salt group (LS group, 20 mmol/L NaCl) and high salt group (HS group, 40 mmol/L NaCl). The viability of VEGFR-3+ cells was measured by CCK-8 method 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). After 24 h and 48 h 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.