甲醇和汽油汽车尾气对A549细胞株细胞毒性及增殖和凋亡功能的影响  

THE STUDY OF PROLIFERATION AND APOPTOSIS OF A549 CELLS ON EXPOSURE TO BOTH GASOLINE AND METHANOL EXHAUST.

作　　者：张浩[1] 梁英[1] 曾祥贵[1] 刘勇[2] 李昌全[3] 蔡春华[3] 吴德生[1] 

机构地区：[1]四川大学华西公共卫生学院环境卫生教研室,成都610041 [2]中国石油西南油气田分公司 [3]中国石油西南油气田分公司西北气矿

出　　处：《现代预防医学》2005年第7期721-723,共3页Modern Preventive Medicine

摘　　要：目的:比较甲醇燃料汽车尾气和汽油燃料汽车尾气对A549细胞株的细胞毒性及增殖和凋亡功能的影响.方法:1)MTT比色法检测受试物细胞毒性.2)受试物体外培养实验:汽油组剂量分组为:0.125 L/ml,0.062 5 L/ml,0.03125L/ml,0.015 6 L/ml,0.007 8 L/ml和一个空白对照;甲醇组剂量分组与汽油相同.A.细胞培养介质乳酸脱氢酶活性的测定;B.细胞形态学观察;C.流式细胞术a.检测细胞周期并计算细胞增殖指数;b.观察细胞亚二倍体峰的变化,计算凋亡百分比;结果:1)MTT比色法汽油组在0.125 L/ml剂量组OD值与对照组相比差异有统计学意义(P＜0.05);甲醇组在1.0L/ml剂量组OD值与对照组相比差异有统计学意义(P＜0.05);2)受试物体外培养实验:A.细胞培养介质乳酸脱氢酶活性的测定:汽油组在0.0625～0.125 L/ml各剂量组LDH值与对照组相比差异有统计学意义(P＜0.05),同等剂量下甲醇组与对照组相比,差异无统计学意义(P＞0.05);B.细胞形态学观察:汽油组在0.125 L/ml,可见大量死亡细胞和凋亡细胞数增加,相同剂量甲醇组未见明显改变;C.流式细胞术:仅汽油组0.015 6,0.031 25L/ml、0.062 5 L/ml剂量组细胞增殖指数显著低于对照组(P＜0.05),汽油组和甲醇组在0.015 6～0.062 5 L/ml剂量组凋亡百分比均高于对照组,但后者作用更强.结论:在同等剂量下,汽油燃料尾气对A549细胞株的毒性及对细胞增殖的抑制明显高于甲醇,但甲醇诱导A549细胞凋亡的作用略强.Objective:To compare the adverse effects of the gasoline exhaust and methanol exhaust on proliferation and apoptosis in A549 cells.Methods:1) The cytotoxicity was detected by MTT assay.2)An in vitro study using A549 cells was done when samples both gasoline and methanol exhaust was given to medium at concentration of 0,0.0078,0.0156,0.03125,0.0625 and 0.125 L/ml.A.LDH in medium was assay. B. cytomorphology was analyzed by light microscope. C.flowcytometry: cell proliferation was determined by both counting proliferation index and analyzing cell cycle . Apoptosis was determined by flow cytometry to observe hypodiploid .and counting percentage of apoptosis cell on different exposure groups .Results:1)The group of gasoline: there was significant difference between control and 0.125 L/ml (P<0.05); the group of methanol: there was significant difference between control and 1.0 L/ml (P<0.05); 2) A.LDH: the group of gasoline: there was significant difference between control and 0.0625-0.125 L/ml (P<0.05); B. Cytomorphology observation: compared with control,the counting of apoptosis and death cell at 0.125 L/ml inceresed significantly; C. Flowcytometry: the group of gasoline: there was significant different PI and percentage of apoptosis between control and above dose (P<0.05); the group of methanol: there was significant difference between control and 0.0625 L/ml (P<0.05); but there was significant different percentage of apoptosis between control and all dose(P<0.05) in the groups of methanol.Conclusion:Both the toxicity and toward the restraining to proliferation in A549 cells of gasoline exhaust is distinctly stronger than the effects of methanol exhaust; But to the apoptosis ,group methanol is greater than group gasoline a little.

关 键 词：A549细胞 汽油 甲醇 增殖与凋亡 

分 类 号：R122.7[医药卫生—环境卫生学]