维生素D对PDCD4表达的调控作用研究

Study on vitamin D in regulating PDCD4 expression in BMDMs

目的:探索维生素D对促炎因子程序性细胞死亡蛋白-4(PDCD4)的调控作用及其分子机制。方法:在巨噬细胞中采用不同剂量维生素D刺激,并在不同时间点进行检测,用实时定量PCR和蛋白质印迹方法确定维生素D刺激对PDCD4 mRNA和蛋白水平的影响,并进一步对PDCD4上游调控因子微小RNA-21(miR-21)进行检测,探索维生素D调节PDCD4的作用机制。最后在脂多糖诱导的炎症细胞模型中,通过蛋白质印迹方法验证维生素D在炎症状态下对PDCD4的调控作用。结果:在巨噬细胞中,维生素D的刺激能够上调miR-21表达,抑制PDCD4表达,而且在炎症模型中,维生素D的抑制作用更为显著。结论:维生素D通过上调miR-21抑制促炎症因子PDCD4的表达,为维生素D抑制炎症的发生提供新的理论依据。

Objective： To elucidate the anti-inflammatory molecular mechanism of vitamin D in regulating the expression of proinflammatory cytokine programmed cell death-4（PDCD4）.Methods： Macrophages were stimulated with different doses of vitamin D and detected at different time points.The expression levels of PDCD4 mRNA and protein were determined using real-time quantitative PCR（q-PCD） and Western blot.The expression of microRNA-21（miR-21） was detected with q-PCR to explore the mechanism of vitamin D in regulating PDCD4.LPS was used to induce inflammation in macrophages to verify the regulating effect of vitamin D on PDCD4 in inflammatory state.Results： Vitamin D inhibited the expression of PDCD4 via upregulating the expression of miR-21, and the inhibitory effect of vitamin D was more significant in the LPS induced inflammation model.Conclusion： Vitamin D inhibits the expression of proinflammatory factor PDCD4 via upregulating miR-21, which may provide a new theoretical basis for the role of vitamin D in inhibiting the occurrence of inflammation.