苯并(a)芘对体外培养人淋巴细胞免疫活性和HSP70的影响

Effects of benzo(a)pyrene on cell immune activity and HSP70 of human lymphocytes in vitro

不同浓度苯并(a)芘[B(a)P](0.125、0.250、0.500、1.000、2.000、4.000、8.000、16.000μmol/L)处理体外培养的人淋巴细胞系721.221,研究B(a)P对体外培养的人淋巴细胞免疫活性和热休克蛋白70的影响.染毒16h,分别应用MTT比色法、Elisa法和Western blot法研究B(a)P对体外培养的淋巴细胞细胞活力、细胞因子IFN-α、interleukin-1α和interleukin-6分泌和热休克蛋白70表达情况的影响.与对照组比较,0.125～1.000μmol/L低剂量B(a)P染毒组促进细胞的生长(P<0.05),1.000μmol/L B(a)P染毒组细胞的增殖活性达到峰值,与溶剂对照组相比,增加了55%.2.000～16.000μmol/L B(a)P染毒细胞,16h后对细胞活力存在显著抑制作用(P<0.05).IFN-α随着B(a)P染毒浓度的升高,IFN-α分泌呈先上升后下降的趋势,1μmol/L B(a)P染毒组分泌达到峰值,随后降低.IL-1α和IL-6分泌随着染毒浓度的增加而降低,与对照组相比有统计学意义(P<0.05),呈现剂量-效应关系.HSP70随着染毒浓度的增加,呈现先增加后降低的趋势.本研究证实B(a)P对721.221细胞活力、免疫活性和应急因子的影响存在显著的剂量依赖性,为今后评估人群接触B(a)P对免疫系统和自身应急系统的毒性提供实验依据.

Human lymphocytes(721.221)were treated with benzo(a)pyrene[B(a)P]at different doses(0.125,0.250,0.500,1.000,2.000,4.000,8.000,16.000μmol/L)for 16hto study the cell viability,imroune activity and HSP70expression in vitro.Cells at exponential growth phase in vitro planted in 96wells microplates were treated by low doses of 0.125-1.000μmol/L and high doses of 2.000-16.000μmol/L B(a)P for 16hrespectively.MTT assay was used to evaluate the effects of B(a)P on cell viability of cells,and the supernatants were collected and analyzed by Elisa to quantify the release of IL-1α,IL-6and IFN-α from the cells,Western blot was used to measure the expression of HSP70.The proliferation of cells were increased at the concentrations of 0.125-1.000μmol/L(P<0.05).The proliferation reached to the highest point at the concentration of 1.000μmol/L.It was enhanced by fifty-five percent compared with the solvent control group.Compared with the negative control,B(a)P at doses of 2.000-16.000μmol/L for 16hon cell viability was significantly inhibited(P<0.05).IFN-αsecretion was first increased and then a downward trend with B(a)P increased.The secretion reached to the highest point at the concentration of 1.000μmol/L,then lowered.IL-1αand IL-6secretion reduced with B(a)P increasing,compared with the control group was statistically significant(P<0.05)in a dose-effect relationship.HSP70expression increased B(a)P increased extend in 0.125-1.000μmol/L groups and then a downward trend with B(a)P increased.Effects of B(a)P on cell viability,immune activity and HSP70 were significantly dependent on the doses.Providing the experimental basis for future assessments of population exposure to B(a)P on the immune system toxicity and emergency response system.