【摘要】 目的:观察抗胆碱药物戊乙奎醚(PHC)对脂多糖(LPS)所介导的A549细胞作用的影响,并探讨其作用机制。方法:采用具备人正常肺泡Ⅱ型细胞特点的A549细胞作为研究对象,实验共分为3组:(1)空白对照组(C组):A549细胞+培养液;(2)LPS组(L组):A549细胞+培养液+10mg/L LPS;(3)LPS+PHC组(L+P组):A549细胞+培养液+10mg/L LPS+5mg/L PHC。培养24h后分别进行以下检测和观察:(1)噻唑蓝(MTT)法药物敏感试验分别检测6h、12h、24h和36h各组A549细胞的吸光度值(OD值);(2)检测各组培养液中损伤因子乳酸脱氢酶(LDH)、碱性磷酸酶(ALP)、丙二醛(MDA)表达水平;(3)用显微镜观测各组细胞形态学改变;(4)应用流式细胞仪检测各组A549细胞周期及细胞凋亡情况。结果:(1)MTT法检测结果:与C组比较,L组和L+P组的OD值明显降低(P<0.05);L+P组与L组比较,L+P组的OD值除6h外均明显升高(P<0.05)。(2)损伤因子检测结果:L+P组LDH和MDA的表达较L组明显降低(P<0.05)。(3)细胞形态学改变:L组A549细胞较C组形态变圆,体积缩小,胞质变空亮,且胞质内出现大小不等的颗粒;加入PHC干预后的L+P组损伤的A549细胞表现有所改善。(4)细胞周期及细胞凋亡情况:与C组比较,L组细胞增殖活性明显下降,细胞凋亡率明显增加,S期细胞比率降低,G0/G1期细胞比率增加;与L组比较,L+P组细胞存活率升高,细胞凋亡率下降,G0/G1期细胞比率降低,G2/M期细胞比率增加。结论:抗胆碱药物PHC能够抑制LPS对A549细胞的损伤,PHC可能参与了A549细胞损伤的保护机制。更多还原

【Abstract】 Objective:To investigate the effects of anticholinergic drug(Penehyclidine(PHC))on lipopolysaccharide(LPS)mediated A549 cells,and to explore its mechanism.Methods:Using A549 cells characterized by normal human alveolar type II cells as research objects,and divided into three groups:(1)Control group(C):DMEM containing A549cells;(2)DMEM mixed with LPS at a concentration of 10mg/L containing A549cells(L);(3)DMEM mixed with LPS at a concentration of 10mg/L and PFC at a ratio(v/v)of 10% containing A549cells(L+C).After cultivated for 24 hours,we carried on the following tests and observations:(1)OD value of A549 cells using MTT(MTT)were detected at 6h,12 h,24h,36h;(2)detected the expression of damage factor of lactate dehydrogenase(LDH),alkaline phosphatase(ALP),and malondialdehyde(MDA);(3)Observed morphological changes in each group;(4)Used flow cytometry to detect the cell cycle and apoptosis of A549 cells.Results:(1)MTT test results:Compared with C group,OD values of L and L+P group was significantly lower.Compared with L group,OD value of the L+P group increased significantly except for the 6hvalue;(2)damage factor test results:the expression of lactate dehydrogenase(LDH)and malondialdehyde(MDA)in L+P group was significantly lower than that in L group;(3)compared with C group,morphology of A549 cells in L group were more rounded,smaller;empty cytoplasm became brighter,and the particles of various sizes occur within the cytoplasm;after PHC intervention,damaged A549 cells in L+P group had been improved;(4)cell cycle and apoptosis:compared with C group,cell proliferation was significantly lower than that in L group.We observed that he apoptosis rate increased,S phase cells decreased,and G0/G1 phase of the cell ratio increased.Compared with L group,L+P group cell viability increased,apoptosis rate decreased,G0/G1 phase cells decreased,and G2/M phase cells increased.Conclusion:Anticholinergics Penehyclidine(PHC)could inhibit lipopolysaccharide(LPS)on A549 cell damage.PHC might involve in the protective mechanism of A549 cell damage.更多还原