IL-10对HaCaT细胞中VEGF-A表达的影响

发布时间：2018-05-31 18:10

本文选题：HaCaT细胞 + IL-　；参考：《安徽医科大学学报》2013年08期

【摘要】：目的探讨白介素-10(IL-10)对皮肤角质形成(HaCaT)细胞中血管内皮生长因子A(VEGF-A)表达的影响及其可能机制。方法培养HaCaT细胞,用不同剂量IL-10刺激后,通过荧光定量PCR(Real-time PCR)在mRNA水平上分析IL-10对HaCaT细胞中VEGF-A表达的影响;通过Western blot法分析IL-10对HaCaT细胞信号通路:丝裂原蛋白激活激酶-胞外信号调节激酶(MAPKs-ERK1/2)及磷脂酰肌醇激酶-丝氨酸/苏氨酸激酶(PI3K-AKT)的影响,并通过MAPKs-ERK1/2特异性阻断剂PD98059及PI3K-AKT特异性阻断剂LY294002阻断这两个信号途径,进而分析IL-10影响HaCaT细胞中VEGF-A表达的可能机制。结果 Real-time PCR分析结果显示IL-10能抑制HaCaT细胞中VEGF-A转录水平的表达;Western blot法结果显示:IL-10能够增加ERK1/2和AKT的磷酸化水平。阻断剂研究显示IL-10抑制VEGF-A转录水平表达的作用不依赖MAPKs-ERK1/2和PI3K-AKT途径。结论 IL-10抑制HaCaT细胞中VEGF-A转录水平的表达,不依赖MAPKs-ERK1/2和PI3K-AKT途径发挥作用。
[Abstract]:Objective to investigate the effect of interleukin-10 (IL-10) on the expression of vascular endothelial growth factor (VEGF-A) in human keratinocytes (HaCaT) and its possible mechanism. Methods HaCaT cells were cultured and stimulated with different doses of IL-10. The effect of IL-10 on VEGF-A expression in HaCaT cells was analyzed at mRNA level by fluorescence quantitative PCR(Real-time. The effects of IL-10 on the signal pathway of HaCaT cells: mitogen-activated kinase-extracellular signal-regulated kinase MAPKs-ERK1 / 2) and phosphatidylinositol kinase-serine / threonine kinase (PI3K-AKT) were analyzed by Western blot assay. The two signaling pathways were blocked by MAPKs-ERK1/2 specific blocker PD98059 and PI3K-AKT specific blocker LY294002, and the possible mechanism of IL-10 affecting VEGF-A expression in HaCaT cells was analyzed. Results Real-time PCR analysis showed that IL-10 could inhibit the expression of VEGF-A transcription in HaCaT cells. Western blot assay showed that WIL-10 could increase the phosphorylation of ERK1/2 and AKT. Antagonist studies have shown that the inhibition of VEGF-A transcription by IL-10 does not depend on the MAPKs-ERK1/2 and PI3K-AKT pathways. Conclusion IL-10 inhibits the expression of VEGF-A in HaCaT cells and does not depend on the MAPKs-ERK1/2 and PI3K-AKT pathways.
【作者单位】：安徽医科大学生物化学教研室;合肥师范学院化学化工系;
【基金】：国家自然科学基金(编号:81271748)
【分类号】：R392.11

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