半乳糖凝集素3对内皮细胞生长、迁移及炎症反应的影响

发布时间：2018-05-02 22:53

本文选题：半乳糖凝集素 + 内皮细胞　；参考：《中国病理生理杂志》2017年06期

【摘要】：目的:探讨半乳糖凝集素3(GAL-3)对人脐静脉内皮细胞(HUVECs)生长、迁移及炎症反应的影响及其作用机制。方法:体外培养HUVECs,给予GAL-3重组蛋白2 mg/L或GAL-3短发夹RNA(shRNA)慢病毒载体处理,实验分为正常对照组、GAL-3重组蛋白处理组、阴性对照组和GAL-3-shRNA组。通过实时荧光定量PCR检测GAL-3、单核细胞趋化蛋白1(MCP-1)、IL-6、基质金属蛋白酶9(MMP-9)和cyclin D1的mRNA水平;利用Western blot检测GAL-3、MCP-1和IL-6的蛋白表达;利用ELISA检测MCP-1和IL-6在培养基中的水平;通过CCK-8实验和划痕实验检测HUVECs的活力和迁移能力;利用Western blot检测信号分子热休克蛋白90(HSP90)、ERK1/2、p-ERK1/2、JNK和p-JNK的蛋白水平。结果:首先,给予GAL-3重组蛋白处理后,GAL-3、MCP-1和IL-6的mRNA及蛋白水平,MMP-9和cyclin D1的mRNA水平,MCP-1和IL-6在培养基的分泌水平均明显高于正常对照组;而GAL-3-shRNA感染细胞后,上述分子的表达水平均低于正常对照组和阴性对照组。其次,GAL-3重组蛋白处理后,细胞活力和迁移能力明显高于正常对照组;而GAL-3-shRNA感染后,细胞活力和迁移能力均低于正常对照组和阴性对照组。此外,GAL-3重组蛋白处理组中,p-ERK1/2和HSP90的蛋白水平高于正常对照组;而GAL-3-shRNA组中,p-ERK1/2和HSP90的蛋白水平均低于正常对照组和阴性对照组。p-JNK的蛋白水平在各组间比较,差异无统计学显著性。结论:GAL-3促进血管内皮细胞的生长、迁移及炎症反应的发生,其机制可能与HSP90-ERK1/2信号通路有关。
[Abstract]:Aim: to investigate the effects of galactose agglutinin 3 (GAL-3) on the growth, migration and inflammatory response of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were cultured in vitro and treated with GAL-3 recombinant protein 2 mg/L or GAL-3 short hairpin RNA-shRNAs. The cells were divided into normal control group, negative control group and GAL-3-shRNA group. The mRNA levels of GAL-3, monocyte chemoattractant protein 1 (MCP-1), matrix metalloproteinase-9 (MMP-9) and cyclin D1 were detected by real-time fluorescence quantitative PCR, the expression of Western blot and IL-6 were detected by Western blot, the levels of MCP-1 and IL-6 were detected by ELISA. The activity and migration ability of HUVECs were detected by CCK-8 test and scratch test, and the protein levels of ERK1 / 2 p-ERK1 / 2 + JNK and p-JNK were detected by Western blot. Results: first of all, the levels of mRNA and protein, the levels of MMP-9 and cyclin D1 of MCP-1 and IL-6 and the levels of mRNA of MMP-9 and cyclin D1 of GAL-3 recombinant protein were significantly higher than those of the normal control group, but the secretion levels of MCP-1 and IL-6 in the culture medium were significantly higher than those in the control group. The expression level of these molecules was lower than that of normal control group and negative control group. After treated with GAL-3 recombinant protein, the cell viability and migration ability were significantly higher than those in the normal control group, while the cell viability and migration ability after GAL-3-shRNA infection were lower than those in the normal control group and the negative control group. In addition, the protein levels of p-ERK1 / 2 and HSP90 in the GAL-3 recombinant protein treatment group were higher than those in the normal control group, while the protein levels of p-ERK1 / 2 and HSP90 in the GAL-3-shRNA group were lower than those in the normal control group and the negative control group. Conclusion the growth, migration and inflammatory response of vascular endothelial cells were promoted by WGAL-3, and the mechanism may be related to the HSP90-ERK1/2 signaling pathway.
【作者单位】：青岛大学医学院附属烟台毓璜顶医院心内科;青岛大学医学院附属烟台毓璜顶医院生物芯片实验室;青岛大学医学院附属烟台毓璜顶医院中心实验室;
【基金】：国家自然科学基金资助项目(No.81571636) 山东省自然科学基金资助项目(No.ZR2015HM058) 山东省医药卫生发展计划资助项目(No.2015WS0017) 烟台市科技发展计划项目(No.2013WS224;No.2015WS021)
【分类号】：R543.5

【相似文献】