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目的探讨Cr(Ⅵ)对细胞内电压依赖性离子通道(VDAC1)mRNA表达和三磷酸腺苷(ATP)水平的干涉效应及其之间的联系。方法实验共分成6个处理组,分别用0、2、4、8、16和32μmol/L Cr(Ⅵ)染毒处理12、24和36 h,然后用逆转录荧光定量聚合酶链反应(RT-qPCR)和荧光素生物发光法分别检测细胞内VDAC1mRNA和能量ATP水平。结果 (1)细胞内VDAC1 mRNA表达在12h明显低于对照组水平,在24h表达水平有所增加,染毒36h后,各剂量组均明显增加,平均增加至对照组的2.65倍;(2)细胞内ATP含量在12h增高,高剂量组(2μmol/L)尤为明显,在24h后细胞内ATP水平明显下降,染毒36h后,低剂量(2μmol/L和4μmol/L)组ATP含量又回升至对照水平,在高浓度(8、16和32μmol/L)组,ATP仍处较低水平;(3)相关分析显示,细胞内VDAC1 mRNA表达与ATP水平之间呈中度负相关(r=0.604,P<0.05)。结论 Cr(Ⅵ)对细胞内ATP水平的干涉效应与VDAC1 mRNA表达异常有关,VDAC1 mRNA表达增加是Cr(Ⅵ)诱导细胞内能量ATP水平降低的分子机制之一。
Objective To investigate the interference effect of Cr (Ⅵ) on the expression of intracellular voltage-dependent ion channel (VDAC1) mRNA and the level of adenosine triphosphate (ATP) and its relationship. Methods The experiment was divided into 6 treatment groups and treated with 0, 2, 4, 8, 16 and 32 μmol / L Cr (Ⅵ) for 12, 24 and 36 h, respectively, and then analyzed by reverse transcription polymerase chain reaction -qPCR) and fluorescein bioluminescence were used to detect intracellular VDAC1 mRNA and energy ATP levels, respectively. Results (1) The expression of VDAC1 mRNA in the cells was significantly lower than that in the control group at 12h, and increased at 24h. After 36h exposure, the expression of VDAC1 mRNA in all the groups increased significantly (2.65 folds of the control) The content of ATP in the cells increased at 12h, especially in the high-dose group (2μmol / L). The content of ATP in the cells decreased obviously after 24h, and the content of ATP in the low dose (2μmol / L and 4μmol / L) (3) Correlation analysis showed that there was a moderate negative correlation between intracellular expression of VDAC1 mRNA and ATP level (r = 0.604, P <0.05). Conclusion The interference effect of Cr (Ⅵ) on intracellular ATP level is related to the abnormal expression of VDAC1 mRNA. The increase of VDAC1 mRNA expression is one of the molecular mechanisms of Cr (Ⅵ) -increasing intracellular ATP level.