炎症条件下毛状样蛋白1促进淋巴细胞与血管内皮细胞的黏附及迁移

Coactosin-Like Protein-1 Promotes Adhesion and Migration of Lymphocytes to Vascular Endothelial Cells under Inflammatory Conditions

目的:研究毛状样蛋白1(Cotl1)在炎症条件下调节淋巴细胞迁移的分子机制。方法:CRISPR/Cas9敲除小鼠胰岛血管内皮细胞MS1的Cotl1基因;黏附实验比较淋巴细胞与野生细胞株和敲除细胞株在LPS刺激下的黏附能力;RT-PCR及流式细胞术比较野生细胞株和敲除细胞株在LPS刺激下表面黏附分子ICAM1、VCAM1、P-selectin的表达情况;Transwell模型比较淋巴细胞在LPS刺激下穿过野生细胞株和敲除细胞株的迁移能力。结果:敲除细胞株Cotl1^-/-构建成功;在LPS刺激下,淋巴细胞与野生细胞株MS1的黏附显著高于敲除细胞株Cotl1^-/-;同时,野生细胞株MS1表面黏附分子ICAM1、VCAM1、P-selectin的表达显著高于敲除细胞株Cotl1^-/-;淋巴细胞穿过野生细胞株MS1的能力显著高于敲除细胞株Cotl1^-/-。结论:炎症条件下Cotl1促进淋巴细胞与血管内皮细胞的黏附及迁移。

Objective:To study the molecular mechanism of coactosin-like protein-1(Cotl1)regulating lymphocytes migration under inflammatory conditions.Methods:The Cotl1 gene of mouse islet vascular endothelial cell lines MS1 was knocked out by CRISPR/Cas9 method.Adhesion assay was used to compare the adhesion ability of 780 lymphocytes to wild cell lines and knockout cell lines under LPS stimulation.RT-PCR and flow cytometry were used to compare the expression of surface adhesion molecules ICAM1,VCAM1 and P-selectin between wild cell lines and knockout cell lines stimulated by LPS.Transwell model was used to compare the transmigration ability of lymphocytes to pass through wild cell lines and knockout cell lines under LPS stimulation.Results:We successfully constructed the knockout cell lines Cotl1^-/-.Under the stimulation of LPS,the adhesion of lymphocytes to the wild cell line MS1 was significantly higher than that of the knockout cell line Cotl1^-/-.Meanwhile,the expression of adhesion molecules ICAM1,VCAM1 and P-selectin on the surface of wild cell lines MS1 was significantly higher than that of knockout cell lines Cotl1^-/-.Finally,the ability of lymphocytes to pass through wild cell lines MS1 was significantly higher than that of knockout cell lines Cotl1^-/-.Conclusion:Cotl1 promotes the adhesion and migration of lymphocytes to vascular endothelial cells under inflammatory conditions.