用含不同浓度维生素C(0,50,100,200,400和800 μmol/L)的培养液培养异育银鲫原代肝脏细胞,待细胞融合后,测定细胞活性、细胞内维生素C含量,乳酸脱氢酶(LDH)活性。再将用维生素C培养的肝脏细胞经敌百虫胁迫24 h,测定细胞内总抗氧化能力(T-AOC)、谷胱甘肽-S-转移酶(GST)和丁酰胆碱酯酶(B-ChE)活性以及细胞内细胞色素P450(CYP450)含量。结果表明,与未添加维生素C组相比,在100 μmol/L的维生素C剂量组,肝脏细胞活性显著高于较其它剂量组(P<0.05);细胞内维生素C的含量随着培养液中维生素C的增加而增加,且差异显著(P<0.05);在800 μmol/L维生素C剂量组,细胞LDH活性显著增加(P<0.05),但是其它剂量组均无显著变化。肝脏细胞经敌百虫胁迫后,在50、100和200 μmol/L维生素C剂量组,细胞T-AOC能力,GST活性,B-ChE活性和CYP450含量显著升高(P<0.05),细胞解毒和抗氧化能力增强;但是当维生素C的剂量为400和800 μmol/L时,细胞T-AOC能力和GST活性降低。综上所述,在体外细胞培养液中添加在50~200 μmol/L剂量范围的维生素C可以促进原代肝脏细胞生长,增强细胞解毒能力,提高细胞的抗氧化水平。

In the present study,the effect of ascorbic acid on primary cultured hepatocytes viability and antioxidant capability under trichlorfon stress were investigated in Carassius auratus gibelio. The hepatocytes were cultured with media containing 0,50,100,200,400 and 800 μmol/L concentration ascorbic acid.Cell viability,the changes in hepatocytes lactate dehydrogenase(LDH)activity and ascorbic acid concentration were assayed.In addition,cellular intracellular total antioxidant capacity(T-AOC),glutathione-S-ePoxide transferase(GST)and butyrylcholinesterase(B-ChE)activities and cytochrome P450(CYP450)concentration under trichlorfon stress were determined.The results showed that hepatocyte viability was significantly increased(P<0.05)when cell was cultured with 100 μmol/L concentrations ascorbic acid in vitro compared with control.Cellular ascorbic acid concentration was significantly increased with media contained 50,100,200,400 and 800 μmol/L concentration ascorbic acid.There were no significant changes in cellular LDH activity with media containing 50,100,200,400 μmol/L concentration ascorbic acid.But LDH activity was markedly(P<0.05)increased when ascorbic acid concentration was 800 μmol/L.Under trichlorfon tress,the cellular antioxidant capability was increased significantly(P<0.05)at 50,100 and 200 μmol/L concentration ascorbic acid treatments.In conclusion,ascorbic acid facilitated hepatocytes viability and function of anti-oxidative stress when cells were cultured with 50-200 μmol/L concentrations ascorbic acid.

现代农业产业技术体系建设专项(CARS-46-20);江苏省科技支撑计划(BE2010394)