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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。
Objective To investigate the effect of ghrelin on the migration of RAW264.7-derived foam cells and its related mechanisms. Methods The oil red O was used to detect the foam cell model. The contents of total cholesterol (TC), free cholesterol (FC) and cholesteryl ester (CE) in foam cells were measured by cholesterol oxidase method. The expression of Akt, p-Akt and cleaved Caspase-3 protein were detected by Western blot. The expressions of p-Akt and cleaved Caspase-3 were detected by immunofluorescence. The change of cytoskeleton was detected by fluorescent cytoskeleton fluorescence probe. To investigate whether PI3K-specific inhibitor LY294002 affects the migration ability of RAW264.7-derived foam cells and the expression of related proteins. Results Treatment of RAW264.7-derived foam cells with 10-7 mol / L ghrelin promoted the migration of foam cells, which could be reversed by LY294002. Western blot results showed that 10-7mol / L ghrelin significantly increased the expression of p-Akt in RAW264.7-derived foam cells and decreased the expression of cleaved Caspase-3 (P <0.05), and significantly improved the expression of RAW264.7-derived foam cells (P <0.05), LY294002 can reverse the above changes. Immunofluorescence staining showed that Akt was significantly expressed in RAW264.7 cells, ghrelin group increased, LY294002 group decreased significantly. Conclusion ghrelin can promote the migration of RAW264.7-derived foam cells, and its molecular mechanism may be related to the activation of PI3K / Akt signaling pathway.