全氟化碳对脂多糖诱导Ⅱ型肺泡上皮损伤的保护机制研究

The protective effect of perfluorocarbon on the injury of alveolar epithelial cells induced by lipopolysaccharide

目的 探讨全氟化碳（perfluorocarbon,PFC）对脂多糖（lipopolysaccharide,LPS）作用下胎鼠Ⅱ型肺泡上皮细胞的凋亡以及炎症反应的影响。方法 分离纯化原代胎鼠Ⅱ型肺泡上皮细胞,采用随机数字表法随机分为对照组、LPS组、PFC组、PFC＋ LPS组.LPS组培养液加入1μg/ml的LPS,PFC组培养液加入20％容积比的PFC,PFC＋ LPS组培养液加入1μg/ml LPS和20％容积比PFC,共同培养后lh后,电镜观察Ⅱ型肺泡上皮细胞的形态学变化,流式细胞仪鉴定各组细胞凋亡率,用ELISA方法检测各组细胞培养上清液中白细胞介素（interleukin,IL）-6及IL-10的浓度.结果 LPS组Ⅱ型肺泡上皮细胞凋亡率（14.29±1.93）％显著高于对照组（10.89±1.04）％,差异有统计学意义（P＜0.05）;PFC＋ LPS组Ⅱ型肺泡上皮细胞凋亡率（12.22±1.47）％显著低于LPS组（P ＜0.05）;LPS组IL-6浓度[（482.58 ±26.84） pg/ml]显著高于对照组[（229.40 ±7.61） pg/ml] （P ＜0.05）,加入PFC后可显著减少LPS诱导的IL-6合成[（265.44 ±29.95） pg/ml] （P ＜0.05）;LPS组IL-10浓度[（1 497.29±191.89） pg/ml]显著高于对照组[（725.87 ±51.83） pg/ml] （P ＜0.05）,加入PFC对LPS诱导的IL-10合成无影响（P＞0.05）。结论 PFC可有效减轻脂多糖诱导的胎鼠Ⅱ型肺泡上皮细胞的炎性损伤,并减轻炎症反应的程度。

Objective To explore the effects of perfluorocarbon （PFC） on the damaged type Ⅱ alveolar epithelial cells （AEC Ⅱ） induced by lipopolysaccharide （LPS）,and the apoptosis and inflammatory reaction of AEC Ⅱ induced by LPS.Methods Primary AEC Ⅱ was divided into control group according to the random number table method,LPS group,PFC group and PFC ＋ LPS group.LPS group：LPS （1 μg/ml） was added to cells.PFC group：PFC （20％） was added to cells.PFC ＋ LPS group：PFC （20％） and LPS （1 μg/ml） were added to cells.The apoptotic rate of AEC Ⅱ was detected by flow cytometry.Morphologic change was observed by electron microscope.Concentrations of intedeukin （IL）-6 and IL-10 of supernatant were detected by ELISA.Results Apoptotic rate of AEC Ⅱ remarkably increased in LPS group than in control grouop [（10.89 ± 1.04） ％ vs （14.29 ± 1.93） ％] （P ＜ 0.05）.Compared with LPS group,the apoptotic rate of AEC Ⅱ decreased remarkably in the PFC ＋ LPS group [（12.22 ± 1.47） ％],（P ＜ 0.05）.IL-6 production of AEC Ⅱ significantly increased in LPS group than in control group [（482.58 ± 26.84） vs （229.40 ± 7.61） pg/ml pg/ml] （P ＜ 0.05）,while decreased in PFC ＋ LPS group [（265.44 ± 29.95） pg/ml].IL-10 production of AEC Ⅱ significantly increased in LPS group than in control group [（1 497.29 ±191.89） pg/ml vs （725.87 ±51.83） pg/ml] （P ＜0.05）,while there was no difference between LPS group and PFC ＋ LPS group （P ＞ 0.05）.Conclusion PFC can protect AEC Ⅱ against the injury induced by LPS.PFC can also release the level of inflammatory response.