CAJ | 학술논문

目的研究镍冶炼烟尘对人肺腺癌细胞(A549)损伤的影响及L抗坏血酸的保护作用.方法将A549细胞分为染毒组和L-抗坏血酸干预组.染毒组加由镍冶炼烟尘配制的浓度分别为0.00、6.25、12.50、25.00、50.00、100.00pg/ml的混悬液,干预组在染毒组的基础上加含有L-抗坏血酸(100mmol/L),作用24h.利用MTT法检测细胞存活率.受试物浓度选取0.00、25.00、50.00、100.00μg/ml进行实验时,用于Flou-3荧光探针检测细胞内Ca2+浓度,DCFH-DA检测细胞内活性氧(ROS)含量,实时定量PCR (real time,RT-PCR)法检测细胞内HIF-1α基因表达.结果随受试物浓度的升高,25.00,50.00,100.00 μg/ml染毒组细胞生长抑制率增加,与0μg/ml染毒组比较,差异均有统计学意义(P＜0.05).镍浓度为25.00,50.00,100.00μg/ml时,染毒组Ca2荧光强度分别为727.70±8.62、777.70±5.50、870.30±13.01,干预组Ca2+荧光强度分别为697.30±15.01、731.70±7.76、825.00±10.54;染毒组ROS荧光强度分别为779.70±4.93、824.30±11.68、899.30±3.06,干预组ROS荧光强度分别为751.30±4.50、789.30±5.67、879.70±7.10;染毒组HIF-1α基因相对表达量分别为2.66±0.28、6.45±0.43、27.42±0.44,干预组HIF-1α基因相对表达量分别为2.10±0.06、5.59±0.27、11.62±0.22;干预组Ca2+荧光强度、ROS荧光强度、HIF-1α基因相对表达量均低于染毒组,差异有统计学意义(P＜0.05).结论随镍浓度增加,A549细胞损害增加,L-抗坏血酸对镍引起的细胞损伤有一定保护作用.
목적 연구얼야련연진대인폐선암세포(A549)손상적영향급L항배혈산적보호작용.방법 장A549세포분위염독조화L-항배혈산간예조.염독조가유얼야련연진배제적농도분별위0.00、6.25、12.50、25.00、50.00、100.00pg/ml적혼현액,간예조재염독조적기출상가함유L-항배혈산(100mmol/L),작용24h.이용MTT법검측세포존활솔.수시물농도선취0.00、25.00、50.00、100.00μg/ml진행실험시,용우Flou-3형광탐침검측세포내Ca2+농도,DCFH-DA검측세포내활성양(ROS)함량,실시정량PCR (real time,RT-PCR)법검측세포내HIF-1α기인표체.결과 수수시물농도적승고,25.00,50.00,100.00 μg/ml염독조세포생장억제솔증가,여0μg/ml염독조비교,차이균유통계학의의(P＜0.05).얼농도위25.00,50.00,100.00μg/ml시,염독조Ca2형광강도분별위727.70±8.62、777.70±5.50、870.30±13.01,간예조Ca2+형광강도분별위697.30±15.01、731.70±7.76、825.00±10.54;염독조ROS형광강도분별위779.70±4.93、824.30±11.68、899.30±3.06,간예조ROS형광강도분별위751.30±4.50、789.30±5.67、879.70±7.10;염독조HIF-1α기인상대표체량분별위2.66±0.28、6.45±0.43、27.42±0.44,간예조HIF-1α기인상대표체량분별위2.10±0.06、5.59±0.27、11.62±0.22;간예조Ca2+형광강도、ROS형광강도、HIF-1α기인상대표체량균저우염독조,차이유통계학의의(P＜0.05).결론 수얼농도증가,A549세포손해증가,L-항배혈산대얼인기적세포손상유일정보호작용.
Objective Studying different concentrations of nickel smelting smoke subjects of human lung adenocarcinoma cells (A549) carcinogenic effects,discusses the influence of L-ascorbic acid protection.Methods The A549 cells were divided into experimental and L-ascorbic acid in the intervention group.Plus exposure group concentration of nickel refining dusts were formulated 0.00,6.25,12.50,25.00,50.00,100.00 μg/ml suspension,the intervention group on the basis of the added exposure group containing L-ascorbic acid (100 mmol/L),contact 24 h.Detection of cell viability by MTT assay.When the test substance concentration select 0.00,25.00,50.00,100.00 μg/ml experiment for internal Flou-3 fluorescent probe to detect cell Ca2+ concentration,within DCFH-DA detect intracellular reactive oxygen (ROS) content,real-time quantitative PCR (real time,in the RT-PCR) was used to detect cell HIF-1α gene expression.Results With the increase of concentration,subjects increased cell growth inhibition rate,intracellular Ca2+ concentration increases,ROS content increased,HIF-1α gene expression increased,differences were statistically significant (P＜0.05).After L-ascorbic acid intervention treatment,the results of the intervention group were lower than that of the experimental group,and the difference was statistically significant (P＜0.05),so L-ascorbic acid can effectively protect the nickel exposure damage to cells.Conclusion With subjects following exposure to nickel concentration increased,its effect on A549 cell damage increases,L-ascorbic acid cell damage caused by nickel has certain protective effect.