Zhang Guixin, Zhang Jingwen, Shang Dong, Qi Bing, Chen Hailong
Department of General Surgery, First Affiliated Hospital of Dalian Medical University, Zhongshan Road 222, Dalian, China, 116011.
Dalian Medical University, Dalian, China, 116044.
In Vitro Cell Dev Biol Anim. 2015 Sep;51(8):851-6. doi: 10.1007/s11626-015-9907-x. Epub 2015 May 20.
The objective of this study is to investigate the effect of deoxycholic acid (DCA) on rat pancreatic acinar cell line AR42J and the functional mechanisms of DCA on AR42J cells. AR42J cells were treated with various concentrations of DCA for 24 h and also treated with 0.4 mmol/L DCA for multiple times, and then, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to detect the AR42J cell survival rate. Flow cytometric was used to detect the cell apoptosis and necrosis in AR42J cells treated with 0.4 mmol/L and 0.8 mmol/L DCA. The cells treated with phosphate buffer saline (PBS) were served as control. In addition, the DNA-binding activity assays of transcription factors (TFs) in nuclear proteins of cells treated with DCA were determined using Panomics Procarta Transcription Factor Assay Kit. The relative survival rates were markedly decreased (P < 0.05) in a dose- and time-dependent manner. Compared with control group, the cell apoptosis and necrosis ratio were both significantly elevated in 0.4 mmol/L DCA and 0.8 mmol/L DCA groups (P < 0.01). A significant increase (P < 0.05) in the activity of transcription factor 2 (ATF2), interferon-stimulated response element (ISRE), NKX-2.5, androgen receptor (AR), p53, and hypoxia-inducible factor-1 (HIF-1) was observed, and the activity of peroxisome proliferator-activated receptor (PPAR), activator protein 1 (AP1), and E2F1 was reduced (P < 0.05). In conclusion, DCA inhibited proliferation and induced apoptosis and necrosis in AR42J cells. The expression changes of related genes regulated by TFs might be the molecular mechanism of AR42J cell injury.
本研究的目的是探讨脱氧胆酸（DCA）对大鼠胰腺腺泡细胞系AR42J的作用及其对AR42J细胞的功能机制。将AR42J细胞用不同浓度的DCA处理24小时，并用0.4 mmol/L DCA多次处理，然后，进行3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）试验以检测AR42J细胞存活率。采用流式细胞术检测用0.4 mmol/L和0.8 mmol/L DCA处理的AR42J细胞中的细胞凋亡和坏死情况。用磷酸盐缓冲盐水（PBS）处理的细胞作为对照。此外，使用Panomics Procarta转录因子分析试剂盒测定用DCA处理的细胞的核蛋白中转录因子（TFs）的DNA结合活性。相对存活率呈剂量和时间依赖性显著降低（P < 0.05）。与对照组相比，0.4 mmol/L DCA和0.8 mmol/L DCA组的细胞凋亡和坏死率均显著升高（P < 0.01）。观察到转录因子2（ATF2）、干扰素刺激反应元件（ISRE）、NKX-2.5、雄激素受体（AR）、p53和缺氧诱导因子-1（HIF-1）的活性显著增加（P < 0.05），而过氧化物酶体增殖物激活受体（PPAR）、激活蛋白1（AP1）和E2F1的活性降低（P < 0.05）。总之，DCA抑制AR42J细胞增殖并诱导其凋亡和坏死。TFs调控的相关基因的表达变化可能是AR42J细胞损伤的分子机制。
