槟榔碱对泡沫细胞胆固醇流出和ABCA1表达的影响

Effect of Arecoline on Cholesterol Efflux and ATP-Binding Cassette Transporter A1 Expression in RAW 264.7 Macrophage-Derived Foam Cells

目的探讨槟榔碱对氧化型低密度脂蛋白(ox-LDL)诱导的鼠源性巨噬细胞性泡沫细胞形成的影响及可能机制。方法采用体外培养的鼠源性巨噬细胞株作为研究对象,加入ox-LDL(50 mg/L)诱导泡沫细胞,同时加入不同浓度的槟榔碱处理,油红O染色进行形态学观察,高效液相色谱法测定细胞内总胆固醇(TC)、游离胆固醇(FC)和胆固醇酯(CE)的水平,3H标记的胆固醇测定胆固醇流出率。采用实时定量PCR和蛋白印迹法分别检测细胞中三磷酸腺苷结合盒转运体A1(ABCA1)的表达。结果 ox-LDL处理48 h后,与正常巨噬细胞相比,油红O染色阳性细胞显著增加,细胞体积明显增大,形态多呈圆形或不规则形;槟榔碱(10-6、10-5和10-4mol/L)组油红O染色阳性细胞数比泡沫细胞模型组显著减少,细胞体积明显缩小。与泡沫细胞模型组相比,槟榔碱(10-5和10-4mol/L)显著性降低细胞内TC、FC、CE水平及CE/TC,显著性增加细胞胆固醇流出率和ABCA1的表达。结论槟榔碱抑制ox-LDL诱导的鼠源性巨噬细胞性泡沫细胞形成,上调泡沫细胞中ABCA1的表达。

Aim To explore the effect of arecoline （Are） on the formation of foam cells induced by the oxidized low density lipoprotein （ox-LDL） and analyze the possible mechanisms. Methods RAW 264.7 macrophages were incubated by ox-LDL （50 mg/L） for 48 h to induce foam cells. Arecoline （10-6mol/L, 10-Smol/L and 10-4mol/L） was added for 48 h at the same time. The cellular lipid accumulation was examined by oil red staining. The cellular con- tents of total cholesterol （TC） and free cholesterol （FC） were detected by high performance liquid chromatography assays. Cholesterol efflux from macrophages was examined by 3H labed cholesterol. The expressions of ATP-binding cassette transporter AI （ABCA1） were detected by Real-time PCR and Western blot. Results Oil red-staining positive ceils were found and macrophages were filled with lipid droplet in foam cells model group. Compared with the foam cells, arecoline （ 10 -5 and 10 44 mol/L） significantly decreased the amount of oil red-staining positive cells and the contents of lipid droplet of foam cells. Compared with thecontrol maerophages group, the contents of TC, FC, cholesteryl ester （CE） and CE/TC ratio in model group were significantly increased in foam cells model group. Arecoline （10-5 and 10-4mol/L）de- creased the contents of TC, FC, cholesteryl ester （CE） and CE/TC ratio, and increased significantly cholesterol efflux and expression of ABCAI compared with foam cells model group. Conclusion Arecoline inhibits the formation of foam cell induced by ox-LDL and up-regulates of expression ABCA1 in RAW 264. 7 macrophage-derived foam cells.