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[目的]研究一氧化氮合酶(NOS)抑制剂干预前后软骨藻酸(DA)对PC12细胞存活率及其产生一氧化氮(NO)的影响。[方法]MTT法检测PC12细胞的存活率;Greiss法测定PC12细胞培养基上清中的NO含量。L-NAME干预前,MTT实验分为对照组及0.0001、0.001、0.01、0.1、1.0μmol/L DA处理组;NO含量测定时分为对照组及0.01、0.1、1.0μmol/L DA处理组。L-NAME干预时,MTT实验及NO含量测定均分为对照组,单纯1.0μmol/L DA处理组,250、1000、2500μmol/L L-NAME+1μmol/L DA处理组。[结果]随着DA染毒剂量的升高,PC12细胞的存活率逐渐下降;0.01、0.1、1.0μmol/L DA作用PC12细胞24 h后,细胞培养基上清NO水平均升高,差异有统计学意义,P<0.05;L-NAME浓度为1000、2500μmol/L干预时,PC12细胞存活率增加,NO生成减少。[结论]DA对PC12细胞具有毒性作用,可以降低细胞的存活率,诱导细胞NO生成增加;NOS抑制剂具有一定保护作用,可明显抑制DA诱导PC12细胞NO生成增加。
[Objective] To investigate the effects of domoic acid (DA) on the survival rate and nitric oxide (NO) production of PC12 cells before and after the intervention of nitric oxide synthase (NOS) inhibitor. [Methods] The survival rate of PC12 cells was detected by MTT method. The content of NO in the supernatant of PC12 cells medium was measured by Greiss method. Before L-NAME intervention, the MTT experiment was divided into control group and DA group, 0.0001,0.001,0.01,0.1,1.0μmol / L DA group; NO concentration was divided into control group and 0.01,0.1,1.0μmol / L DA group. L-NAME intervention, MTT assay and NO content were divided into control group, 1.0μmol / L DA group, 250,1000,2500μmol / L L-NAME group and 1μmol / L DA group. [Results] The survival rate of PC12 cells decreased gradually with the increase of DA dose. After treated with 0.01, 0.1 and 1.0 μmol / L DA for 24 h, the levels of NO in supernatant of cell culture medium increased Statistical significance, P <0.05; L-NAME concentration of 1000,2500μmol / L intervention, PC12 cell survival increased, NO production decreased. [Conclusion] DA has a toxic effect on PC12 cells, which can reduce the survival rate of cells and induce the increase of NO production. NOS inhibitor has a certain protective effect, and can significantly inhibit the increase of NO production induced by DA in PC12 cells.