论文部分内容阅读
目的探讨增塑剂邻苯二甲酸二丁酯对类神经细胞PC12细胞增殖能力的影响及其对PC12细胞凋亡相关基因表达的调控,为接触人群可能产生远期效应的生物学检测提供参考评价指标。方法体外培养类神经细胞株PC12细胞,培养基分别含有0,12.5,25,50 mg/L邻苯二甲酸二丁酯,作用24,48,72和96 h后,用细胞计数CCK-8法检测细胞的存活和增殖,台盼蓝染色绘制生长曲线,DNA凝胶电泳观察DNA-Ladder,RT-PCR检测p53,bcl-2,Bax等凋亡相关基因表达变化情况。结果邻苯二甲酸二丁酯在12.5 mg/L时即表现出显著的抑制PC12细胞增殖的作用(P<0.01),且随剂量增加抑制作用增强,同时出现DNA断裂阳性;凋亡相关基因均有表达,其中p53基因、Bax基因表达增加,bcl-2基因表达降低,且呈剂量相关关系(P<0.01)。结论邻苯二甲酸二丁酯暴露可抑制PC12细胞增殖并诱导PC12细胞凋亡,同时通过调节凋亡相关基因p53、Bax、bcl-2基因的表达水平而在其凋亡发生中起重要作用。推测邻苯二甲酸二丁酯长期低剂量暴露的累积效应与阿尔茨海默病以及其他神经细胞凋亡相关性疾病的发生发展可能存在相关关系。
Objective To investigate the effect of plasticizer dibutyl phthalate on the proliferation of PC12 cells and the regulation of apoptosis related genes in PC12 cells so as to provide reference for the biological detection of long-term effects index. Methods The neural stem cell line PC12 was cultured in vitro. The medium contained 0, 12.5, 25 and 50 mg / L dibutyl phthalate, respectively. After 24, 48, 72 and 96 h, The survival and proliferation of cells were detected by trypan blue staining, and the growth curve was drawn. The expression of p53, bcl-2, Bax and other apoptosis-related genes were detected by DNA gel electrophoresis and DNA-Ladder. Results The results showed that dibutyl phthalate showed a significant inhibitory effect on the proliferation of PC12 cells at a dose of 12.5 mg / L (P <0.01), and its inhibitory effect was enhanced with the increase of the dose. (P <0.01). The p53 gene and Bax gene expression were increased and the expression of bcl-2 gene was decreased in a dose-dependent manner (P <0.01). Conclusion Dibutyl phthalate exposure can inhibit the proliferation of PC12 cells and induce the apoptosis of PC12 cells, and play an important role in the apoptosis of PC12 cells by regulating the expression of p53, Bax and bcl-2 genes. It is speculated that the cumulative effect of long-term low-dose dibutyl phthalate exposure and Alzheimer’s disease and other neuronal apoptosis-related diseases may be related to the occurrence and development.