SjCystatin通过PI3K-Akt通路诱导调节性巨噬细胞产生VEGF-C

SjCystatin upregulated VEGF-C production in regulatory macrophage via PI3K-Akt pathway activation

搞要:目的探讨一种可诱导抑炎性巨噬细胞分化的虫源性分子(SjCystatin)对巨噬细胞内血管内皮生长因子C(VEGF-C)产生的影响机制。方法将巨噬细胞分为对照组、脂多糖(LPS)处理组、LPS+免疫复合物(LPS+IC)处理组、白细胞介素-4(IL-4)/10处理组、SjCystatin处理组和通路抑制剂处理组,每组10只BALB/C小鼠。应用荧光定量PCR的方法检测各组巨噬细胞分化标志性分子和VEGF-C mRNA水平的变化,应用Western blot和ELISA方法检测各处理组VEGF-C产生的变化。结果 SjCystatin处理组巨噬细胞内LIGHT、IL-10等调节性巨噬细胞标志分子较LPS处理组分别升高2.1倍和5倍,差异有统计学意义(P<0.05);而CCL-17、CXCL-13等M2型巨噬细胞标志分子低于IL-4/10处理组,差异有统计学意义(P<0.05)。SjCystatin处理组巨噬细胞上清、细胞内VEGF-C表达升高,差异有统计学意义(P<0.05)。SjCystatin处理组巨噬细胞内p-Akt水平显著高于LPS处理组,当PI3K-Akt通路抑制剂加入后可显著降低VEGF-C的产生。结论调节性巨噬细胞产生VEGF-C亚型,PI3K-Akt信号通路在巨噬细胞产生VEGF-C的过程中发挥重要作用。

Objective To investigate the effect of SjCystatin on the production of vascular endothelial growth factor C(VEGF-C)in macrophages. Methods Macrophages were divided into lipopolysaccharide(LPS)treated group,LPS+IC treated group,IL-4/10 treated group,SjCystatin-treated group and PI3K inhibitor treated group,10 BALB/C mice in each groups.Quantitative PCR was used to detect the changes of macrophage differentiation markers and VEGF-C mRNA levels. Western blot and ELISA were used to detect the changes of VEGF-C production in each group. Results The levels of LIGHT and IL-10(markers of regulatory macrophage)in SjCystatin-treated group were 2.1 and 5 times higher than those in LPS-treated group(P<0.05),but the levels of CCL-17 and CXCL-13 were significantly lower than those in IL-4/10-treated group(P<0.05). The expression of VEGF-C in macrophage supernatant and cell lysates were significantly increased in SjCystatintreated group(P<0.05). The level of p-Akt in SjCystatin-treated group was significantly higher than that of LPS-treated group. When PI3K-Akt pathway inhibitor was added,the production of VEGF-C in SjCystatin-treated group was significantly reduced. Conclusion Regulatory macrophages were the source of VEGF-C;PI3K-Akt signaling pathway might play an important role in the process of VEGF-C production in macrophages.