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。
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 A549 cells; （2） DMEM mixed with LPS at a concentration of 10 mg/L containing A549 cells （L） ; （3） DMEM mixed with LPS at a concentration of 10 mg/L and PFC at a ratio （v/v） of 10% containing A549 cells （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 6 h,12 h,24 h, 36 h; （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 6h value; （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, damagedA549 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.
出处
《广西医科大学学报》
CAS
2016年第1期10-14,共5页
Journal of Guangxi Medical University
基金
广西自然科学基金资助项目(No:2013GXNSFBA019182)
关键词
戊乙奎醚
脂多糖
A549
penehyclidine
lipopolysaccharide
A549 cells
