口灌醋酸棉酚对鲤血清转氨酶、肝脏抗氧化酶活力及肝细胞凋亡的影响

Effects of Gossypol Acetate on Serum Aminotransferase,Liver Antioxidant Enzyme Activities and Hepatocyte Apoptosis of Cyprinus carpio following Oral Administration

为了评价醋酸棉酚对鲤肝脏的急性毒性效应,对鲤单次口灌20 mg/kg醋酸棉酚,分别于口灌后0.5、1、2、4、6、12、24、48、72 h时测定鲤血清中谷草转氨酶(GOT)、谷丙转氨酶(GPT)和肝脏超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活力;观察口灌醋酸棉酚24 h时,鲤肝细胞的组织学特征和凋亡现象,并检测鲤肝脏中caspase3、caspase 6、caspase 7基因表达水平。结果显示,对鲤单次口灌20 mg/kg醋酸棉酚后,其血清GOT活力在0.5 h时显著高于对照组未口灌醋酸棉酚的鲤(P<0.05),在48、72 h时显著低于对照组(P<0.05),在1、2、4、6、12、24 h时与对照组相比无显著性差异(P>0.05);其血清GPT活力在0.5、1 h时显著高于对照组(P<0.05),4、48 h时显著低于对照组(P<0.05),2、6、12、24、72 h时与对照组相比无显著性差异(P>0.05)。鲤肝脏SOD活力在0.5、1、2、48 h时显著高于对照组(P<0.05),4、12 h时显著低于对照组(P<0.05),6、24、72 h时与对照组相比无显著性差异(P>0.05);GSH-Px活力在0.5、1、2、24、48、72 h时显著高于对照组(P<0.05),6、12 h时显著低于对照组(P<0.05),4 h时与对照组相比无显著性差异(P>0.05)。口灌醋酸棉酚24 h时,鲤肝细胞体积和细胞核大小无明显变化,细胞未发生凋亡现象;鲤肝脏中caspase3、caspase 6、caspase 7基因表达量均显著低于对照组(P<0.05)。综上,对鲤单次口灌20 mg/kg醋酸棉酚后,鲤的肝功能短时间内受到了一定程度影响,但未引起组织病变或细胞凋亡,可能是通过激活机体抗氧化酶系统缓解了醋酸棉酚的毒性作用。

In order to evaluate the acute hepatotoxicity of gossypol acetate in Cyprinus carpio , the activities of serum gluamic-oxalacetic transaminase (GOT),glutamic pyruvic transaminase (GPT), hepatic superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were analyzed at 0.5,1,2,4,6,12,24,48,72 h after oral administration of gossypol acetate at20 mg/kg.The histological characteristics of liver, TUNEL of hepatocytes, and the genes expression of caspase3, casspase 6 , caspase 7 were analyzed at24 h after oral administration.The results showed that the serum GOT activity of20 mg/kg dose group was significantly higher than the control group at 0.5 h ( P <0.05),and it was significantly lower at 48 h and 72 h than the control group ( P <0.05),and there was no significant difference compared with the control group at1,2,4,6,12,24 h ( P >0.05).The serum GPT activity was significantly higher than the control group at 0.5 h and1 h ( P <0.05),it was lower than the control group at4h and 48 h ( P <0.05),and there was no significant difference compared with the control group at2,6,12,24,72 h ( P >0.05).The activity of SOD in liver was significantly higher than the control group at 0.5 ,1,2,48 h ( P <0.05),significantly lower at4h and12 h than the control group ( P <0.05),and there was no significant difference compared with the control group at 6,24,72 h ( P > 0.05 ).The GSH-Px activity in liver was significantly higher than that of the control group at 0.5,1,2,24,48,72 h ( P < 0.05 ),and significantly lower than the control group at 6 h and12 h( P <0.05), and there was no significant difference between the gossypol group and the control group at4h ( P >0.05).There was no significant change in the volume of hepatocytes and nucleus,and there was no apoptosis in the cells of Cyprinus carpio at24 h after oral administration.The expression levels of caspase3,caspase 6 and caspase 7 in liver were significantly lower than those in the control group ( P <0.05).In summary,the results suggested that the liver function was affected in a short time following oral administration,but the liver was not damaged, and the free gossypol did not induce apoptosis in liver cells of Cyprinus carpio . Cyprinus carpio may increase its own antioxidant enzyme activities to reduce the toxic effects of gossypol acetate.