摘要
目的 探讨聚苯乙烯纳米塑料(Polystyrene nanoplastics,NPs)对大鼠主动脉平滑肌细胞(Rat aortic smooth muscle cells,RASMCs)泡沫化的影响.方法 体外培养大鼠主动脉平滑肌细胞,用不同水平的直径为80纳米的绿色荧光NPs(80 nm NPs)处理RASMCs 24小时;采用细胞计数试剂盒(Cell counting kit-8,CCK8)检测细胞活力,并检测细胞释放的乳酸脱氢酶(Lactate dehydrogenase,LDH)活性以评估细胞损伤程度;利用 DiI 荧光标记的氧化型低密度脂蛋白(Dioctadecyl-3,3,3,3-tetramethylin docarbocyanine-oxidized low density lipoprotein,DiI-oxLDL)诱导平滑肌细胞泡沫细胞形成,并行油红O染色显示脂滴;采用蛋白免疫印迹法检测自噬相关指标(Microtubule-associated protein 1 light chain 3B,MAP1LC3B)的表达水平.结果 80 nm NPs可被RASMCs吞噬并产生细胞毒性,呈水平依赖性促进RASMCs摄取oxLDL,自噬相关指标(LC3B)表达水平降低.结论 聚苯乙烯纳米塑料可被血管平滑肌细胞吞噬并促进泡沫细胞形成,其潜在机制可能是通过抑制RASMCs自噬来实现的.
Abstract
Objective To investigate the role of polystyrene nanoplastics(NPs)in foam cell formation in rat aortic smooth muscle cells(RASMCs).Methods RASMCs were treated with green fluorescent NPs of 80-nanometer diameter(80 nm NPs)at different concentrations for 24 hours.Cell viability was assessed using the CCK-8 assay.Lactate dehydrogenase(LDH)activity assay kit was used to measure LDH release as an indica-tor of cell damage.Foam cell formation was induced using DiI-labeled oxidized low-density lipoprotein(DiI-ox-LDL).Oil Red O staining was performed to visualize lipid droplets.Protein immunoblotting was conducted to assess the expression levels of autophagy-related marker microtubule-associated protein 1 light chain 3B(MAP1LC3B/LC3B).Results The results demonstrated that RASMCs internalized the 80 nm NPs,and their uptake was associated with concentration-dependent cytotoxicity.Furthermore,the NPs promoted the uptake of oxLDL by RASMCs and downregulated the expression level of the autophagy-related marker LC3B.Conclu-sion The 80nm NPs promote the uptake of oxLDL by RASMCs and induce foam cell formation.The potential mechanism underlying this effect may involve the inhibition of autophagy in RASMCs.
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