| Object: To investigate the protective effects of perfluorocarbon (PFC) onhuman alveolar epithelial-like cells A549injured by lipopolysaccharide (LPS)from the perspective of cell biology.Materials and Methods: Human alveolar epithelial-like cells A549weresubcultured conventionally and divided into four groups:(1) control group: cellsdid not receive any intervention;(2) PFC group: PFC was added to the cell culturemedium to a final volume concentration (vol/vol, PFC: culture media) of10%.After ultrasonic mixing, the mixed liquor containing PFC and culture media wastransferred into the culture dish;(3) LPS group: cells were incubated with LPS ata final concentration of100μg/ml;(4) LPS+PFC group (co-culture group): cellswere incubated with both LPS and PFC at above concentration, respectively. Theexperiment was investigated in following four aspects:(1) Morphological changeswere observed under inverted microscope and photographed after24h,48h and72h of treatment respectively.(2) A549cells were seeded in culture plate atcertain numbers. The cell numbers of each group were counted using cell countingmethod every day after dosing for6days, and then cell growth curve was drew.(3)A549cells were mechanically wounded with a pipette tip before givencorresponding treatment. The time point of administration was defined as0h. Thearea of the denuded surface was measured immediately after administration andagain after24h and48h respectively. The percentage of wound healing andinternuclear distance was analyzed with image analysis software imageJ (NIHImage1.55).(4) The apoptotic rates of A549cells of four groups were examined by flow cytometry24hours after administration.Result:(1) Cell morphology of LPS group were sparser and smallercompared with other three groups of cells, part of the cells were round. The cellmorphology of remaining three groups was full and arranged closely.(2) The cellgrowth curve showed that cell proliferation of LPS group was inhibited obviously(P<0.05).(3) Wound healing assay showed that the percentage of wound healingat48h of control group, LPS group, PFC group and LPS+PFC group were42.8%,25.3%,37.0%and33.2%, respectively. Differences in percentage of woundhealing between LPS group and the other three groups all reached statisticalsignificance (p<0.05).The internuclear distance at scratch edge of control group,LPS group, PFC group and LPS+PFC group were25.2μm,20.7μm,24.9μm and24.5μm, respectively. Differences in internuclear distance between LPS group andthe other three groups all reached statistical significance (p<0.05).(4) Theapoptosis rate of A549in control group, LPS group, PFC group and LPS+PFCgroup were5.33%,35.76%,5.22%and13.39%, respectively. Compared withcontrol, PFC and LPS+PFC group, the apoptosis rate in LPS group wassignificantly increased (p<0.01). There was no statistic difference between thecontrol and PFC group. Compared with LPS group, the apoptosis rate inLPS+PFC group was significantly decreased (p<0.01).Conclusion:(1) PFC could alleviate the adverse effect of LPS on themorphology and growth status of A549cells conspicuously.(2) PFC couldalleviate the adverse effect of LPS on proliferation of A549cells markedly.(3)PFC could improve wound healing of A549significantly which was inhibited byLPS. The improving effect of PFC on A549in wound healing ability was relativeto its protective role on cell migration.(4) PFC could alleviate LPS inducedapoptosis significantly. |
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