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[目的]研究三(2-氯乙基)磷酸酯[tris(2-chloroethyl)phosphate,TCEP]对肝细胞线粒体功能的影响。[方法]用0.00(溶剂对照组)、3.12、12.50、50.00和200.00 mg/L TCEP分别处理人正常肝细胞(L02细胞)和人肝癌细胞(Hep G2细胞)24 h和48 h。测定细胞活力、线粒体内活性氧水平、线粒体DNA拷贝数、线粒体膜电位和胞内游离钙(Ca2+)水平以及胞内ATP的浓度。[结果]与相应溶剂对照组相比,在24 h,200.00 mg/L TCEP处理组L02细胞活力降低(P<0.05),≥12.50 mg/L TCEP处理组Hep G2细胞活力降低(P<0.05);在48 h,50.00 mg/L和200.00 mg/L TCEP处理组两种细胞活力降低(P<0.05)。所有TCEP处理组两种细胞的线粒体DNA拷贝数均减少(P<0.05或P<0.01)。在24 h,所有TCEP处理组两种细胞的线粒体膜电位均下降(P<0.05或P<0.01)。在24 h,200.00 mg/L TCEP处理组两种细胞胞内游离Ca2+水平均升高(P<0.01);在48 h,50.00 mg/L和200.00 mg/L TCEP处理组两种细胞胞内游离Ca2+均升高(P<0.01)。在24 h和48 h,200.00 mg/L TCEP处理组L02胞内ATP水平降低(P<0.05);在24 h,50.00 mg/L和200.00 mg/L TCEP处理组Hep G2胞内ATP浓度下降(P<0.05或P<0.01)。[结论]一定浓度的TCEP可致肝细胞线粒体损伤和线粒体功能指标异常,提示TCEP有肝细胞线粒体毒性。
[Objective] To investigate the effect of tris (2-chloroethyl) phosphate, TCEP on the mitochondrial function of hepatocytes. [Method] Human normal hepatocytes (L02 cells) and human hepatoma cells (Hep G2 cells) were treated with 0.00 (solvent control), 3.12,12.50,50.00 and 200.00 mg / L TCEP for 24 h and 48 h, respectively. Cell viability, mitochondrial ROS level, mitochondrial DNA copy number, mitochondrial membrane potential and intracellular free calcium (Ca2 +) levels and intracellular ATP concentration were determined. [Result] Compared with the corresponding solvent control group, the viability of L02 cells in 200.00 mg / L TCEP treatment group decreased (P <0.05) at 24 h and the viability of Hep G2 cells in ≥ 12.5 mg / L TCEP treatment group decreased (P <0.05) At 48 h, the viability of both cells in the 50.00 mg / L and 200.00 mg / L TCEP treatment groups decreased (P <0.05). Mitochondrial DNA copy number was decreased in all TCEP-treated cells (P <0.05 or P <0.01). At 24 h, mitochondrial membrane potential decreased in all TCEP-treated cells (P <0.05 or P <0.01). At 24 h, intracellular free Ca2 + levels were increased in TCEP-treated cells (200.00 mg / L) (P <0.01) Ca2 + were increased (P <0.01). At 24 h and 48 h, the intracellular ATP level of L02 in 200.00 mg / L TCEP treatment group decreased (P <0.05); at 24 h, the intracellular ATP concentration of Hep G2 in 50.00 mg / L and 200.00 mg / L TCEP treatment groups decreased P <0.05 or P <0.01). [Conclusion] A certain concentration of TCEP can cause mitochondrial damage and mitochondrial dysfunction in hepatocytes, suggesting that TCEP has hepatocyte mitochondrial toxicity.