【摘要】 目的 通过检测脂多糖诱导的MLE-12细胞炎性模型中TNF-α、TLR4、AKT、ERK的表达情况，研究胰岛素减轻脂多糖对小鼠肺上皮细胞炎症反应的分子机制，以期为急性肺损伤的潜在治疗方法提供实验基础和理论依据。方法 脂多糖(10 ug/ml)诱导MLE-12细胞构建炎性肺泡上皮细胞损伤模型，更换血清培养基，胰岛素或PBS处理72 h。将细胞分成5组：control组(C组):MLE-12细胞常规培养基培养；LPS组(L组):加入LPS(10 ug/ml)培养；LPS+胰岛素1组(L+I1组):加入0.1 nM胰岛素处理；LPS+胰岛素2组(L+I2组):加入1 nM胰岛素处理；LPS+胰岛素3组(L+I3组):加入3 nM胰岛素处理。每组分别于12 h、24 h、72 h 3个时间点进行测定。结果 MLE-12细胞在脂多糖(10 ug/ml)作用下，TNF-α、TLR4等细胞炎性因子的表达明显升高。胰岛素的处理可以部分抑制脂多糖对MLE-12细胞造成的炎症反应。胰岛素可以通过抑制PI3K/AKT、MAPK/ERK信号通路中蛋白的磷酸化水平来降低肺上皮细胞的炎症反应。结论 胰岛素可通过调节AKT、ERK磷酸化水平部分减轻脂多糖对肺上皮细胞的炎症反应，这为临床合理应用胰岛素干预治疗ALR、ARDS提供了初步的理论基础。更多还原

【Abstract】 Objective To study the molecular mechanism of insulin attenuating the inflammatory response of lipopolysaccharide on mouse lung epithelial cells by detecting the expression of TNF-α, TLR4, AKT and ERK in the lipopolysaccharide-induced inflammatory model of MLE-12 cells, with a view to providing experimental basis and theoretical basis for potential therapeutic approaches for acute lung injury.Methods MLE-12 cells were used as the research object, induced by adding lipopolysaccharide(10 ug/ml) for 24h, to construct the model of inflammatory alveolar epithelial cell injury, replacing the serum medium, and treated with insulin or PBS for 72h.The cells were divided into 5 groups: control group(C):MLE-12 cells were cultured in regular medium; LPS group(L): MLE-12 cells were cultured by adding LPS(10 ug/ml); LPS+Insulin 1 group(L+I1): which 0.1 nM insulin treatment was added to continue the culture; LPS+Insulin 2 group(L+I2): which 1nM insulin treatment was added to continue the culture; LPS+insulin 3 group(L+I3):which 3nM insulin treatment was added to continue the culture. Each group was measured at three time points, 12h, 24h and 72h, respectively.Results The expression of cellular inflammatory factors such as TNF-α and TLR4 was significantly elevated in MLE-12 cells in the presence of lipopolysaccharide(10 ug/ml).Treatment with insulin partially inhibited the inflammatory response caused by lipopolysaccharide on MLE-12 cells.Insulin can reduce the inflammatory response in lung epithelial cells by inhibiting the phosphorylation levels of proteins in the PI3K/AKT and MAPK/ERK signaling pathways.Conclusion Insulin attenuates the inflammatory response of lipopolysaccharide on alveolar epithelial cells in part by regulating the levels of AKT and ERK phosphorylation.This provides a preliminary theoretical basis for the rational clinical application of insulin intervention in the treatment of ALR/ARDS.更多还原