ghrelin通过PI3K/Akt信号通路促进RAW264.7源性泡沫细胞迁移  

Ghrelin Promotes the Migration of Foam Cells Through PI3K/Akt Signal Pathway

作　　者：丁英鹏 孙玉慧[2] 杜荣增[1] 王中群[1] 严金川[1] 徐绥宁 杨洪强[3] 朱杰[4] 张薪茹 史雷忠 

机构地区：[1]江苏大学附属医院心内科,江苏省镇江市212001 [2]广东江门出入境检验检疫局江门国际旅行卫生保健中心,广东省江门市529000 [3]济宁市第一人民医院急诊科,山东省济宁市272011 [4]六安市人民医院心内科,安徽省六安市237005 [5]溧阳市人民医院心内科,江苏省常州市213300

出　　处：《中国动脉硬化杂志》2016年第5期440-446,共7页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(81370408);江苏省自然科学基金项目(BK20131246);江苏省卫生厅项目(Q201308);镇江市社会发展项目(SH2015038;SH2015023和SH2015036);卫生部核医学重点实验室和江苏省分子核医学重点实验室开放课题(KF201504)

摘　　要：目的探讨ghrelin对RAW264.7源性泡沫细胞迁移的影响及相关机制。方法油红O检测泡沫细胞模型的构建,胆固醇氧化酶法检测泡沫细胞内总胆固醇(TC)、游离胆固醇(FC)和胆固醇酯(CE)含量,transwell小室实验检测ghrelin对RAW264.7源性泡沫细胞迁移的影响,Western blot检测Akt、p-Akt和cleaved Caspase-3蛋白的表达,免疫荧光检测p-Akt和cleaved Caspase-3的表达,细胞骨架荧光探针检测细胞骨架的变化。观察PI3K特异性抑制剂LY294002是否影响RAW264.7源性泡沫细胞的迁移能力及相关蛋白的表达。结果 10-7mol/L ghrelin处理RAW264.7源性泡沫细胞可以促进泡沫细胞迁移,此过程可以被LY294002逆转。Western blot结果显示10-7mol/L ghrelin可显著升高RAW264.7源性泡沫细胞p-Akt的表达,降低cleaved Caspase-3的表达(P<0.05),并明显改善RAW264.7源性泡沫细胞的迁移能力(P<0.05),LY294002可逆转以上变化。免疫荧光检测显示Akt在RAW264.7细胞明显表达,ghrelin组表达增多,LY294002组明显降低。结论 ghrelin可促进RAW264.7源性泡沫细胞迁移,其分子机制可能与激活PI3K/Akt信号通路有关。Aim The effects of ghrelin on RAW264. 7 derived foam cells migration and its associated mechanisms were investigated. Methods The detection of total cholesterol（ TC）,free cholesterol（ FC）,cholesterol ester（ CE） and lipid droplets were performed by cholesterol oxides method and oil red O staining,respectively. The effects of ghrelin on RAW264.7 derived foam cells migration were detected by transwell chamber assay,the expression of Akt,p-Akt,cleaved Caspase-3 were semi-quantified by Western blot,the distribution of p-Akt and cleaved Caspase-3 in cells was qualitatively analyzed with fluorescent staining. The dissociation and polymerization of cell cytoskeleton was observed with F-actin fluorescence probe. Results 10-7mol / L ghrelin could promote the migration of RAW264.7 derived foam cells,which was reversed by LY294002. Western blot analysis showed that 10-7mol / L ghrelin could significantly increase the expression of p-Akt and reduce the expression of cleaved Caspase-3（ P0.05） in RAW264.7 derived foam cells.What＇s more,it could improve the ability of RAW264.7 migration obviously（ P0.05）. Interestingly,these processes above can be reversed by LY294002 treatment. Fluorescent analysis demonstrated that there was an obvious expression of Akt in RAW264.7 cells. The treatment of ghrelin could upregulate its expression,while LY294002 treatment downregulated the expression. Conclusion Ghrelin could promote the migration of RAW264.7 derived foam cells,which may be related to the activation of the PI3 K / Akt signal pathway.

关 键 词：细胞迁移 泡沫细胞 GHRELIN PI3K/AKT信号通路 

分 类 号：R363[医药卫生—病理学]