摘要
目的：探讨低剂量脂多糖（Lipopolysaccharide,LPS）对PC12细胞抗氧化能力的影响。方法：分别以浓度为0、0.01、0.025、0.05、0.1、0.25、0.5、1μg/μl的LPS刺激PC12细胞24小时,MTT法检测细胞活性确立对细胞没有毒性作用的安全剂量;采用安全剂量刺激PC12细胞24小时,收集培养基上清和细胞,采用试剂盒检测上清中的总抗氧化能力（T-AOC）,细胞中超氧化物歧化酶（SOD）的表达,Western blot检测细胞内Bcl-2蛋白的表达;流式细胞仪检测细胞内活性氧（ROS）、钙离子（Ca2＋）的平均荧光强度。结果：LPS浓度小于0.1μg/μl对细胞没有明显毒性作用（P〉0.05）;采用0、0.01、0.025、0.05、0.1μg/μlLPS分别刺激PC12细胞24小时,培养上清中T-AOC呈下降趋势,以0.05、0.1μg/μl组明显（P〈0.01）,细胞内SOD呈上升趋势（0.05μg/μl组,P〈0.05;0.1μg/μl组,P〈0.01）,细胞内Bcl-2表达逐渐升高（0.0250.1μg/μl组,P〈0.01）,细胞内ROS呈下降趋势（0.1μg/μl组,P〈0.01）,钙离子未见明显变化（P〉0.05）。结论：低剂量LPS处理PC12 24小时具有剂量依赖性增强细胞抗氧化能力,并可能藉此参与应激耐受。
Objective：To investigate the effects of low-dose LPS on anti-oxidant capacity in PC12 cells.Methods：Stimulating PC12 cells with various concentrations of LPS（0,0.01,0.025,0.05,0.1,0.25,0.5,1 μg/μl） for 24 h,MTT assay was conducted to establish safe doses for cell viability.Non-lethal doses of LPS were used to deal with PC12 cells for 24 h,the culture supernatant and cells were collected.The total anti-oxidant capacity in culture supernatant and superoxide dismutase in cells were detected with assay kits.The intracellular Bcl-2 protein expressions were detected with Western-blot,and the average fluorescence intensity of ROS and calcium were tested with flow cytometry.Results：Less than 0.1 μg/μl LPS concentration had no toxicity for cell activity（P〉0.05）.Stimulating PC12 cells with different concentrations of LPS（0,0.01,0.025,0.05,0.1 μg/μl）,the T-AOC decreased significantly,especially in 0.05 μg/μl and 0.1 μg/μl groups（P〈0.01） and the intracellular SOD increased（0.05 μg/μl group,P〈0.05;0.1 μg/μl group,P〈0.01）.The Bcl-2 expressions in PC12 cells increased gradually（0.025-0.1 μg/μl group,P〈0.01）.The intracellular ROS decreased（0.1 μg/μl group,P〈0.01）,but there was no significant changes in calcium（P〉0.05）.Conclusion：Low-dose LPS increases PC12 cells＇ anti-oxidant capacity at 24 h in a dose-dependent manner,and that probably participates in stress tolerance.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2011年第5期430-433,共4页
Chinese Journal of Immunology
