环磷酰胺诱导的尼罗罗非鱼体内外肝损伤模型的构建  

Establishment of the acute hepatic injury model induced by cyclophosphamide in tilapia(Oreochromis niloticus) in vivo and in vitro

作　　者：曹丽萍[1,2] 杜金梁[1,2] 贾睿[1,2] 丁炜东[1,2] Galina Jeney 徐跑[1,2] 殷国俊[1,2] CAO Liping;DU Jinliang;JIA Rui;DING Weidong;Galina Jeney;XU Pao;YIN Guojun(Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization,Ministry of Agriculture and Rural Affairs,Freshwater Fisheries Research Center,Chinese Academy of Fishery Sciences,Wuxi 214081,China;International Joint Research Laboratory for Fish Immunopharmacology,Chinese Academy of Fishery Sciences,Wuxi 214081,China;Research Institute for Fisheries,Aquaculture and Irrigation,Ann Light 8,Szarvas H-4440,Hungary)

机构地区：[1]中国水产科学研究院淡水渔业研究中心,农业农村部淡水渔业和种质资源利用重点实验室,无锡214081 [2]中国水产科学研究院淡水渔业研究中心,农业农村部鱼类免疫药理学国际联合实验室,无锡214081 [3]Research Institute for Fisheries,Aquaculture and Irrigation,Ann Light 8,Szarvas H-4440,Hungary

出　　处：《安徽农业大学学报》2022年第1期76-80,共5页Journal of Anhui Agricultural University

基　　金：中央级公益性科研院所基本科研业务费专项资金(2019JBFM11);江苏自然科学基金(BK20201143)共同资助。

摘　　要：以环磷酰胺(CTX)为诱导物,建立罗非鱼体内外急性肝损伤模型。体外,0、5、10、15、20和25 mmol·L^(-1) CTX分别作用于离体培养的罗非鱼精密肝切片(PCLS)6 h后,测定切片培养上清中谷丙转氨酶(GPT)、谷草转氨酶(GOT)和乳酸脱氢酶(LDH)水平,同时收集肝切片,用噻唑蓝(MTT)法检测肝切片增殖活性,测定切片内总抗氧化能力(T-AOC)、丙二醛(MDA)、过氧化物歧化酶(SOD)和谷胱甘肽(GSH)水平。体内,采用胸腔注射法造模,每千克鱼体重分别注射0、10、25、75和100 mg环磷酰胺(下文以mg·kg^(-1)表示),每隔3 d给药1次,连续3次。末次给药后第4天取血,测定罗非鱼血清中GPT、GOT和LDH及肝组织匀浆中T-AOC、MDA、SOD和GSH水平。结果显示:20~25 mmol·L^(-1) CTX作用体外培养精密肝切片6 h及75~100 mg·kg^(-1) CTX胸腔注射罗非鱼后,均可以导致GPT、GOT和LDH活力显著升高;T-AOC、SOD活力及GSH含量显著下降,MDA含量显著提高。同时,20和25 mmol·L^(-1) CTX能导致肝切片增殖活性显著下降,分别为空白对照组的67.29%和55.41%,且有剂量依赖性。结果表明:CTX可引起罗非鱼肝损伤;分别采用20 mmol·L^(-1) CTX作用体外培养PCLS 6 h或者采用75~100 mg·kg^(-1) CTX胸腔注射罗非鱼,每隔3 d给药1次,连续3次,均可以构建体内外急性肝损伤模型。The present study aimed to develop acute liver injury models of tilapia(Oreochromis niloticus)in vivo and in vitro using cyclophosphamide(CTX)as hepatotoxicant.In vitro,the precision cut liver slices(PCLS)were exposed to 6 concentrations of CTX(0,5,10,15,20 and 25 mmol·L^(-1))for 6 h,and the levels of glutamate oxalate transaminase(GOT),glutamate pyruvate transaminase(GPT)and lactate dehydrogenase(LDH)in slices culture supernatant were determined.At the same time,the total antioxidant capacity(T-AOC),superoxide dismutase(SOD),glutathione(GSH)and malondialdehyde(MDA)levels in slices and the survival rate of the PCLS were detected.In vivo,tilapias were injected with 0,10,25,50,75 and 100 mg·kg^(-1) CTX intrapleurally.It was ad-ministered for intrapleurally once every three days for three times.Samples were taken on the 4th day after the last administration.The GPT,GOT and LDH of serum and T-AOC,SOD,GSH and MDA of liver tissue ho-mogenates were detected.The results showed that both in the PCLS treated with CTX at 20-25 mmol·L^(-1) for 6 h and the tilapias injected with 75-100 mg·kg^(-1) CTX,the activities of GPT,GOT and LDH were significantly increased,the MDA production were increased,while the activities of T-AOC,SOD and the GSH content were decreased.Meanwhile,20 and 25 mmol·L^(-1) CTX significantly decreased PCLS viabilities(67.29%and 55.41%,respectively)compared to the control dose-dependently.Overall results proved CTX can cause liver injury in tilapia;PCLS cultured in vitro with 20 mmol·L^(-1) CTX for 6 h or tilapia injected with 75-100 mg·kg^(-1) CTX intrapleurally once every three days for three times can build acute liver injury models in vivo and in vitro,respectively.

关 键 词：环磷酰胺 罗非鱼 肝损伤模型 体内外 

分 类 号：S965.125[农业科学—水产养殖] S941.91[农业科学—水产科学]