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目的:探讨丹参酮ⅡA对氧化型低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的鼠源巨噬细胞性泡沫细胞形成的影响及其机制。方法:体外培养的鼠源性巨噬细胞株,用ox-LDL(50μg/mL)孵育细胞以诱导泡沫细胞,同时用不同质量浓度的丹参酮ⅡA(10-5,10-4和10-3mol/L)处理。用油红O染色观察细胞荷脂情况,高效液相色谱法测定细胞内总胆固醇(total cholesterol,TC)、游离胆固醇(free cholesterol,FC)和胆固醇酯(cholesteryl ester,CE)的水平。采用[3H]标记的胆固醇测定胆固醇流出率。实时定量PCR和Western印迹法分别检测细胞中三磷酸腺苷结合盒转运体A1(ATP-binding cassette transporter A1,ABCA1)的mRNA和蛋白的表达。结果:Ox-LDL处理48 h后,与正常巨噬细胞相比,油红O染色阳性细胞数显著性增加,细胞体积明显增大,细胞内可见大量的脂滴,形态多呈不规则形。与泡沫细胞模型相比,10-4或10-3mol/L丹参酮ⅡA处理组油红O染色阳性细胞数显著减少,细胞内脂滴明显减少,细胞体积明显缩小。与泡沫细胞模型组相比,10-4或10-3mol/L丹参酮ⅡA显著性降低了细胞内TC,FC,CE水平和CE/TC比值,显著性增加了细胞胆固醇流出率和ABCA1mRNA和蛋白的表达。结论:丹参酮ⅡA可抑制ox-LDL诱导的鼠源巨噬细胞性泡沫细胞形成,其机制可能与丹参酮ⅡA增加泡沫细胞中ABCA1的表达,促进胆固醇流出有关。
Objective: To investigate the effect of tanshinone ⅡA on the formation of murine macrophage foam cells induced by oxidized low density lipoprotein (ox-LDL) and its mechanism. Methods: Murine macrophages cultured in vitro were incubated with ox-LDL (50μg / mL) to induce foam cells. Tanshinone ⅡA (10-5, 10-4 and 10-3 mol / L). Oil red O staining was used to observe cell lipid-loading. The contents of total cholesterol (TC), free cholesterol (FC) and cholesteryl ester (CE) in the cells were determined by HPLC. Cholesterol efflux was measured using [3H] labeled cholesterol. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expression of ATP-binding cassette transporter A1 (ABCA1). RESULTS: Compared with normal macrophages, the number of Oily Red O-stained cells increased significantly after 48 hours of Ox-LDL treatment, and the volume of cells increased obviously. A large number of lipid droplets were observed in the cells, and the morphology was irregular. Compared with foam cell model, the numbers of oil red O positive cells in 10-4 or 10-3mol / L tanshinone ⅡA treatment group were significantly decreased, the intracellular lipid droplets was significantly reduced, and the cell volume was significantly reduced. Compared with foam cell model group, 10-4 or 10-3mol / L tanshinone ⅡA significantly reduced intracellular TC, FC, CE levels and CE / TC ratio, significantly increased the rate of cellular cholesterol efflux and ABCA1 mRNA and protein expression. CONCLUSION: Tanshinone IIA can inhibit the formation of murine macrophage foam cells induced by ox-LDL. The possible mechanism is that Tanshinone IIA can increase the expression of ABCA1 and promote the efflux of cholesterol in foam cells.