全氟化碳对脂多糖致A549细胞损伤的细胞形态学和增殖能力的影响

Morphological and proliferative effects of perfluorocarbon on lipopolysaccharide-induced injury in A549 cells

目的从细胞生物学角度研究全氟化碳(PFC)对脂多糖(LPS)诱导A549细胞损伤的保护作用,为临床应用PFC治疗急性呼吸窘迫综合征提供理论依据。方法常规传代培养人肺泡上皮样细胞A549,将其分为4组。对照组:不作任何干预;PFC组:按10%体积比(PFC∶培养液)加入全氟辛烷;LPS组:干预时按100mg/L的浓度加入LPS;LPS+PFC组:按上述浓度同时加入LPS和PFC。各组A549细胞在给予相应处理后24、48、72h于倒置显微镜下观察形态学变化并拍照记录。A549细胞按一定细胞数铺板,每组3个复孔,加药后每天应用细胞计数法计数各组细胞数目,共6d,绘制细胞生长曲线。结果(1)细胞形态学显示LPS组较其他3组细胞明显稀疏,皱缩明显,部分细胞呈圆形,生长状态变差;其余3组细胞形态饱满,排列紧密,生长状态良好。(2)细胞生长曲线显示LPS组细胞增殖受抑制,从加药后第4天开始,LPS组与其余各组细胞数目的差异均有统计学意义(P<0.05或P<0.01)。结论PFC能够显著改善LPS对A549细胞形态及生长状态的不良影响,改善LPS对A549细胞增殖的抑制作用。

Objective To investigate the protective effects of perfluorocarbon (PFC) on human alveolar epithelial-like cells A549 injured by lipopolysaccharide (LPS) from the perspective of cell biology, and provide a theoretical basis for the clinical application of PFC in the treatment of acute respiratory distress syndrome. Methods Human alveolar epithelial-like cells A549 were subcultured conventionally and divided into four groups:(1)control group: cells did not receive any intervention.(2)PFC group: PFC was added to the cell culture medium to a final concentration (vol/vol, PFC∶culture media) of 10%.(3)LPS group: cells were incubated with LPS at a final concentration of 100 mg/L.(4)LPS+ PFC group: cells were incubated with both LPS and PFC at above concentration, respectively.Morphological changes were observed under inverted microscope and photographed at the 24th, 48th and 72nd hour after treatment respectively.A549 cells were seeded in culture plate at certain numbers.The cell numbers of each group were counted using cell counting method every day after dosing for six days, and then cell growth curve was drew. Results (1)Cell morphology of LPS group were sparser and smaller compared with other three groups of cells, part of the cells were round.The cell morphology of other three groups was full and arranged closely.(2)The cell growth curve showed that cell proliferation of LPS group was inhibited obviously.There was statistical significance in the cell numbers between LPS group and other three groups since the fourth day after treatment (P<0.05 or P<0.01). Conclusions PFC can markedly alleviate the adverse effect of LPS on the morphology, growth status and proliferation of A549 cells.