维生素D通过维生素D受体-过氧化物酶体增殖物激活受体γ信号通路对高糖诱导的巨噬细胞血管内皮生长因子C表达的影响

Effects of vitamin D on the expression of VDR,PPARγand VEGFC in macrophages induced by high glucose through VDR-PPARγsignaling pathway

目的探讨维生素D(VitD)对高糖诱导的巨噬细胞(MO)内维生素D受体(VDR)、PPARγ、血管内皮生长因子C(VEGFC)表达的影响。方法通过前期浓度及时间依赖实验,以浓度25 mmol/L葡萄糖依赖性培养小鼠MO系(U937)24 h。实验分为对照组(Con)、VitD组、PPARγ抑制剂(T0070907)组、VitD+T0070907组。ELISA法检测TNF-a、IL-6、VEGFC水平,实时荧光定量PCR、Western blot法分别检测VDR、PPARγ、VEGFC基因及蛋白表达水平。结果VitD呈浓度、时间依赖性表达增加MO内TNF-α、IL-6、VEGFC水平,在VitD浓度为0.1 mmol/L作用24 h产生最大效应。与Con组比较,VitD组VDR、PPARγ、VEGFC基因及蛋白表达水平升高(P<0.05)。与VitD组比较,T0070907组、VitD+T0070907组PPARγ、VEGFC基因及蛋白表达水平减少(P<0.05)。结论VitD可能通过VDR-PPARγ信号通路,促使MO内VDR、PPARγ、VEGFC表达增加,减轻高糖环境下MO炎症反应。

Objective To investigate the effect of vitamin D(VitD)on the expression of vitamin D receptor(VDR),peroxisome proliferator activated receptorγ(PPARγ)and vascular endothelial growth factor C(VEGFC)in macrophages(MO)induced by high glucose.Methods Based on the previous concentration and time-dependent experiments,25 mmol/L glucose was selected to culture mouse Mo line(U937)for 24 h.The experiment was divided into control group(Con),Vit D group,T0070907 group and Vit D+T0070907 group.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and VEGFC were detected by ELISA.The expression levels of VDR,PPARγ,VEGFC gene and protein were detected by RT-PCR and Western blot.Results VitD increased the levels of TNF-a,IL-6 and VEGFC in Mo in a concentration and time-dependent manner.The maximum effect was observed at the concentration of 0.1 mmol/L for 24 h.Compared with con group,VDR,PPARγ,VEGFC gene and protein expression were increased in Vit D group(P<0.05).Compared with Vit D group,the expression of PPARγ,VEGFC protein and gene in T0070907 group and Vit D+T0070907 group were decreased(P<0.05).Conclusion VitD may increase the expression of VDR,PPARγand VEGFC in Mo through VDR-PPARγsignaling pathway,and reduce the inflammatory response of Mo in high glucose environment.