IL-10对HaCaT细胞中VEGF-A表达的影响

IL-10 inhibits the expression of VEGF-A in HaCaT cells

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。

Objective To explore the effect of IL-10 on VEGF-A in HaCaT cells. Methods HaCaT cells were clutured and treated with various concentration of IL-10 for different time. Real-time PCR was performed to detect the level of the VEGF-A mRNA; Western blot were used to analyzed signaling pathway activated by IL-10, inclu- ding mitogen-activation protein kinase-extracellular signal-regulated kinase （MAPKs-ERK1/2） and phosphatidyli- nositol kinase-serine/threonine kinases （PI3K-AKT）. The specific inhibitors of the ERK1/2 and AKT phosphoryla- tion were used to detect the mechanism of the effect of IL-10 on the expression of VEGF-A. Results Our results showed that that IL-10 could inhibit the expression of VEGF-A. After IL-10 was stimulated, the IL-10 could in- crease the phosphorylation of ERK1/2 and AKT. The studies of the inhibitors of ERK1/2 （PD98059） and AKT （LY294002） showed the effect of IL-10 on the VEGF-A was independent of the MAPKs-ERK1/2 and PI3K-AKT signaling pathway. Conclusion The IL-10 can inhibit the expression of VEGF-A in HaCaT cells, and this effect of the IL-10 is independent of the MAPKs-ERK1/2 and PI3K-AKT signaling pathway.