【摘要】：目的探討對氨基水楊酸鈉(PAS-Na)對鉛誘導PC12細胞凋亡的影響。方法 PC12細胞培養(yǎng)至其對數(shù)生長期后,正常對照組和正常+PAS-Na 500μmol·L~(-1)組細胞于正常培養(yǎng)液中培養(yǎng)24 h后棄培養(yǎng)液,前者加入正常培養(yǎng)液、后者加入PAS-Na繼續(xù)培養(yǎng)24 h。醋酸鉛模型組及醋酸鉛+PAS-Na 20,100和500μmol·L~(-1)組先與醋酸鉛(終濃度10μmol·L~(-1))共培養(yǎng)24 h后棄培養(yǎng)液,再加入相應濃度PAS-Na繼續(xù)培養(yǎng)24 h。用MTT法測定細胞存活率,試劑盒方法測定細胞內還原型谷胱甘肽(GSH)含量,Hoechst33342熒光染色檢測凋亡率,AnnexinⅤ/PI雙染流式細胞術檢測PC12細胞早期凋亡率,Western蛋白質印跡法檢測Bcl-2、Bax和P53蛋白水平。結果與正常對照組比較,醋酸鉛模型組細胞存活率降低(P0.05),細胞凋亡形態(tài)變化典型,細胞早期凋亡率增高(P0.05),細胞內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)組上述指標未見明顯變化。與醋酸鉛模型組比較,醋酸鉛+PAS-Na組細胞存活率增高(P0.05),細胞凋亡率和早期凋亡率均降低(P0.05),細胞內GSH含量增高(P0.01),P53蛋白水平和Bax/Bcl-2比值降低(P0.05)。結論 PAS-Na對鉛致PC12細胞凋亡有一定的拮抗作用,其機制可能與PAS-Na抗脂質過氧化損傷和降低Bax/Bcl-2比值有關。
[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.
【作者單位】： 廣西醫(yī)科大學公共衛(wèi)生學院衛(wèi)生毒理學教研室;
【基金】：國家重點基礎研究發(fā)展計劃(973計劃)(2012-CB525001)~~
【分類號】：R114

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