对氨基水杨酸钠对铅诱导体外培养PC12细胞凋亡的影响
发布时间：2019-07-24 15:39
【摘要】：目的探讨对氨基水杨酸钠(PAS-Na)对铅诱导PC12细胞凋亡的影响。方法 PC12细胞培养至其对数生长期后,正常对照组和正常+PAS-Na 500μmol·L~(-1)组细胞于正常培养液中培养24 h后弃培养液,前者加入正常培养液、后者加入PAS-Na继续培养24 h。醋酸铅模型组及醋酸铅+PAS-Na 20,100和500μmol·L~(-1)组先与醋酸铅(终浓度10μmol·L~(-1))共培养24 h后弃培养液,再加入相应浓度PAS-Na继续培养24 h。用MTT法测定细胞存活率,试剂盒方法测定细胞内还原型谷胱甘肽(GSH)含量,Hoechst33342荧光染色检测凋亡率,AnnexinⅤ/PI双染流式细胞术检测PC12细胞早期凋亡率,Western蛋白质印迹法检测Bcl-2、Bax和P53蛋白水平。结果与正常对照组比较,醋酸铅模型组细胞存活率降低(P0.05),细胞凋亡形态变化典型,细胞早期凋亡率增高(P0.05),细胞内GSH含量降低(P0.01),P53蛋白水平升高(P0.01),Bax/Bcl-2比值升高(P0.01);正常+PAS-Na 500μmol·L~(-1)组上述指标未见明显变化。与醋酸铅模型组比较,醋酸铅+PAS-Na组细胞存活率增高(P0.05),细胞凋亡率和早期凋亡率均降低(P0.05),细胞内GSH含量增高(P0.01),P53蛋白水平和Bax/Bcl-2比值降低(P0.05)。结论 PAS-Na对铅致PC12细胞凋亡有一定的拮抗作用,其机制可能与PAS-Na抗脂质过氧化损伤和降低Bax/Bcl-2比值有关。
[Abstract]:Objective to investigate the effect of sodium p-aminosalicylic acid (PAS-Na) on apoptosis of PC12 cells induced by lead. Methods after PC12 cells were cultured in logarithmic growth phase, normal control group and normal PAS-Na 500 渭 mol 路L ~ (- 1) group were cultured in normal culture medium for 24 h and then abandoned the culture medium. the former was added to normal culture medium, and the latter was added to PAS-Na for 24 h. Lead acetate model group and lead acetate PAS-Na 20100 and 500 渭 mol 路L ~ (- 1) groups were co-cultured with lead acetate (final concentration 10 渭 mol 路L ~ (- 1) for 24 h, then the culture medium was abandoned after 24 h, and then the corresponding concentration of PAS-Na was added to the culture medium for 24 h. The cell survival rate was measured by MTT assay, the content of reduced glutathione (GSH) in cells was measured by kit method, the apoptosis rate was detected by Hoechst33342 fluorescence staining, the early apoptosis rate of PC12 cells was detected by Annexin V / PI double staining flow cytometry, and the levels of Bcl-2,Bax and p53 protein were detected by Western imprinting. Results compared with the normal control group, the survival rate of lead acetate model group was decreased (P 0.05), the morphological changes of apoptosis were typical, the early apoptosis rate was increased (P 0.05), the intracellular GSH content was decreased (P 0.01), the p53 protein level was increased (P 0.01), and the Bax/Bcl-2 ratio was increased (P 0.01). There was no significant change in the above indexes in the normal PAS-Na 500 渭 mol 路L ~ (- 1) group. Compared with the lead acetate model group, the cell survival rate, apoptosis rate and early apoptosis rate, intracellular GSH content, p53 protein level and Bax/Bcl-2 ratio in lead acetate PAS-Na group were higher than those in lead acetate model group (P 0.05), and the apoptosis rate and early apoptosis rate in lead acetate group were significantly lower than those in lead acetate model group (P 0.05). Conclusion PAS-Na has antagonistic effect on apoptosis of PC12 cells induced by lead, and its mechanism may be related to the anti-lipid peroxide injury of PAS-Na and the decrease of Bax/Bcl-2 ratio.
【作者单位】： 广西医科大学公共卫生学院卫生毒理学教研室;
【基金】：国家重点基础研究发展计划(973计划)(2012-CB525001)~~
【分类号】：R114
[Abstract]:Objective to investigate the effect of sodium p-aminosalicylic acid (PAS-Na) on apoptosis of PC12 cells induced by lead. Methods after PC12 cells were cultured in logarithmic growth phase, normal control group and normal PAS-Na 500 渭 mol 路L ~ (- 1) group were cultured in normal culture medium for 24 h and then abandoned the culture medium. the former was added to normal culture medium, and the latter was added to PAS-Na for 24 h. Lead acetate model group and lead acetate PAS-Na 20100 and 500 渭 mol 路L ~ (- 1) groups were co-cultured with lead acetate (final concentration 10 渭 mol 路L ~ (- 1) for 24 h, then the culture medium was abandoned after 24 h, and then the corresponding concentration of PAS-Na was added to the culture medium for 24 h. The cell survival rate was measured by MTT assay, the content of reduced glutathione (GSH) in cells was measured by kit method, the apoptosis rate was detected by Hoechst33342 fluorescence staining, the early apoptosis rate of PC12 cells was detected by Annexin V / PI double staining flow cytometry, and the levels of Bcl-2,Bax and p53 protein were detected by Western imprinting. Results compared with the normal control group, the survival rate of lead acetate model group was decreased (P 0.05), the morphological changes of apoptosis were typical, the early apoptosis rate was increased (P 0.05), the intracellular GSH content was decreased (P 0.01), the p53 protein level was increased (P 0.01), and the Bax/Bcl-2 ratio was increased (P 0.01). There was no significant change in the above indexes in the normal PAS-Na 500 渭 mol 路L ~ (- 1) group. Compared with the lead acetate model group, the cell survival rate, apoptosis rate and early apoptosis rate, intracellular GSH content, p53 protein level and Bax/Bcl-2 ratio in lead acetate PAS-Na group were higher than those in lead acetate model group (P 0.05), and the apoptosis rate and early apoptosis rate in lead acetate group were significantly lower than those in lead acetate model group (P 0.05). Conclusion PAS-Na has antagonistic effect on apoptosis of PC12 cells induced by lead, and its mechanism may be related to the anti-lipid peroxide injury of PAS-Na and the decrease of Bax/Bcl-2 ratio.
【作者单位】： 广西医科大学公共卫生学院卫生毒理学教研室;
【基金】：国家重点基础研究发展计划(973计划)(2012-CB525001)~~
【分类号】：R114
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