α-硫辛酸在微囊藻毒素-LR诱导草鱼卵巢细胞损伤中的作用

Protective role of α-lipoic acid against microcystin-LR-induced grass carp ovary cell damage

为研究α-硫辛酸(Alpha lipoic acid,α-LA)在微囊藻毒素-LR(Microcystin-LR,MC-LR)诱导水生动物毒性效应中的保护作用,本研究以草鱼卵巢(Grass carp ovary,GCO)细胞为研究对象,检测分析了不同浓度α-LA以及α-LA与MC-LR共同暴露对GCO细胞活力的影响,随后根据测定的结果设置对照组(不添加MC-LR和α-LA)、125μmol·L^(-1)α-LA组、24μmol·L^(-1)MC-LR组及125μmol·L^(-1)α-LA+24μmol·L^(-1)MC-LR组,检测分析了α-LA在缓解MC-LR对GCO细胞活性、氧化应激以及炎症方面的影响。结果表明:与对照组相比,24μmol·L^(-1)MC-LR可诱导乳酸脱氢酶(LDH)活性和丙二醛(MDA)含量显著上升,同时显著抑制谷胱甘肽(GSH)活性(P<0.05),当MC-LR处理组中加入125μmol·L^(-1)α-LA后,与MC-LR单独暴露组相比,LDH活性和MDA含量均显著下降(P<0.05),但GSH活性却显著上升(P<0.05)。对氧化相关基因分析发现,与对照组相比,24μmol·L^(-1)MC-LR可显著降低SOD1、CAT和GST基因的表达量(P<0.05),当125μmol·L^(-1)α-LA与24μmol·L^(-1)MC-LR共同作用于GCO细胞后,与MC-LR单独暴露组相比,联合暴露组的GST基因的表达量显著上升(P<0.05),但SOD1和CAT两个基因的表达变化不显著(P>0.05)。此外,对炎症因子进行分析发现,MC-LR单独暴露组TNFα和IL11基因的相对表达水平显著高于对照组(P<0.05),但联合暴露组中的TNFα和IL11基因的相对表达水平却显著低于MC-LR单独暴露组(P<0.05)。研究结果表明,α-LA可缓解MC-LR诱导的氧化应激,提高细胞活力,抑制GCO细胞炎症的发生,从而减弱MC-LR对GCO细胞的损伤。

To evaluate the protective effects of alpha lipoic acid(α-LA)on microcystin-LR(MC-LR)-induced toxicity in aquatic animals,grass carp ovary(GCO)cells were used in this study.The viability of GCO cells across different concentrations of α-LA treatment and the joint exposure ofα-LA with MC-LR were analyzed.Then,the control(without adding MC-LR and α-LA),125μmol·L^(-1)α-LA,24μmol·L^(-1) MC-LR,and 125μmol·L^(-1)α-LA+24μmol·L^(-1) MC-LR groups were set according to these cell viability results;finally,the effects ofα-LA on the cell viability,oxidative stress,and inflammation of MC-LR-induced GCO cells were analyzed.The results showed that 24μmol·L^(-1) MC-LR treatment significantly increased lactic dehydrogenase(LDH)activity and malondialdehyde(MDA)content,and significantly inhibited the glutathione(GSH)activity of the GCO cells compared to that of the control group(P<0.05).When 125μmol·L^(-1)α-LA was added to the MC-LR treated group,LDH activity and MDA content were significantly reduced compared to those of the MC-LR exposed group(P<0.05);however,GSH activity was significantly increased(P<0.05).Compared with the control group,the 24μmol·L^(-1) MC-LR group exhibited a significant lower SOD1,CAT,and GST gene expression(P<0.05).In the 125μmol·L^(-1)α-LA+24μmol·L^(-1) MCLR group,GST gene expression was significantly increased(P<0.05);however,the expressions of SOD1 and CAT genes were not significantly changed compared with those of the MC-LR exposure group(P>0.05).In addition,analysis of inflammatory factors demonstrated that the relative expression levels of TNFα and IL11 genes in the MC-LR exposure group were significantly higher than those in the control group(P<0.05);nonetheless,the relative expression levels of TNFα and IL11 genes in the combined exposure group were significantly lower than those in the MC-LR exposure group(P<0.05).These results indicate that α-LA can alleviate MC-LR-induced oxidated stress,improve cell viability,and inhibit inflammation of GCO cells,and thus reduce MC-LR-mediated damage to GCO cells.