Effects of Vitamin C on Resistance to Dimethoate Stress in Juvenile Schizothorax grahami
YANG Tongzhi1, ZHAN Huixiang2, YAO Junjie1, YANG Mei1, WEI Yulai1, YANG Longbao1
1. College of Animal Sciences, Guizhou University, Guiyang 550025, China; 2. Aquaculture Techniques Extension Station of Bijie, Guizhou Province, Bijie 551700, China
Abstract：40% Dimethoate was added to the mountain spring water in a tank with culture of Schizothorax grahami with body weight of (1.52±0.26) g at a dose of 0 (control group), 5.091, 10.182, 24.89, and 39.598 mg/L (96 h LC50=39.598 mg/L, safe concentration SC=10.182 mg/L) at water temperature of (13.5±0.5) ℃ for 120 h, and then 90 mg/L of vitamin C was added to water body in each group in order to investigate the toxicity of dimethoate to juvenile S. grahami, the toxic environment simulated by injecting dimethoate into the aquaculture water. The changes in activities of Fethoxy resorufin-deethylase (EROD), AchE, and catalase (CAT) and malondialdehyde (MDA) and the expression levels of HSP70 gene in juvenile S. grahami under dimethoate stress were investigated, and the detoxification effect of vitamin C on the stress was further explored. Activities of EROD, acetylcholinesterase and MDA content were analyzed in the muscle, liver, and gills of Schizothorax grahami in each group collected at 0, 24, 48, 96, and 120 h, and the expression of HSP70 mRNA and gene cloning in another muscle samples in a 5 to 10-fold volume of RNA preservation solution to investigate the ability of vitamin C to resist dimethoate stress in juvenile S. graham. The results showed that there were significantly increase in activities of EROD, AchE, and CAT and MDA level in juvenile S. graham in the over SC groups (P<0.05), with the lower activities of EROD, CAT, and AchE and MDA level in the SC group and the less than SC groups after adding vitamin C, the higher activity of EROD, and lower activities of CAT, and AchE and MDA level were observed in the juvenile S. graham exposed to the water containing vitamin C. The expression of HSP70 mRNA was shown to be significantly induced by different concentrations of dimethoate (P<0.05), without significant change in the other groups (P>0.05), except the maximal concentration group from 24 h to 96 h. After 96 h, however, the expression level of HSP 70 gene was decreased significantly in each group (P<0.05), close to the control group in the two lower concentration groups. The findings indicated that the damage to the body by dimethoate was not found to be recovery at dimethoate concentration of over SC, to a certain extent, without effect on the body at dimethoate concentration of less than SC. Vitamin C led to an active role in protecting the body from oxidative damage caused by exogenous substances to a certain extent, even though dimethoate induced the expression level of HSP 70 gene in juvenile S. grahami fish.
杨通枝, 詹会祥, 姚俊杰, 杨梅, 韦玉来, 杨龙保. 维生素C对昆明裂腹鱼幼鱼抗乐果胁迫能力的影响[J]. 水产科学, 2020, 39(6): 871-879.
YANG Tongzhi, ZHAN Huixiang, YAO Junjie, YANG Mei, WEI Yulai, YANG Longbao. Effects of Vitamin C on Resistance to Dimethoate Stress in Juvenile Schizothorax grahami. Fisheries Science, 2020, 39(6): 871-879.
[1]詹会祥,周礼敬,晏宏,等. 昆明裂腹鱼生物学特性和流水养殖技术[J]. 中国水产,2011(9):29-30. [2]赵飞,王摆,张晓娜,等. 久效磷农药对金鱼肝细胞DNA的损伤及其机制研究[J]. 中国环境科学,2015,35(5):1563-1569. [3]Gagnon M M, Holdway D A. EROD induction and biliary metabolite excretion following exposure to the water accommodated fraction of crude oil and to chemically dispersed crude oil[J]. Archives of Environmental Contamination and Toxicology,2000,38(1):70-77. [4]Solé M, Porte C, Albaigés J. The use of biomarkers for assessing the effects of organic pollution in mussels[J]. The Science of the Total Environment,1995,159(2/3):147-153. [5]Solé M. Assessment of the results of chemical analyses combined with the biological effects of organic pollution on mussels[J]. TrAC Trends in Analytical Chemistry,2000,19(1):1-9. [6]Thomas J P, Maiorino M, Ursini F, et al. Protective action of phospholipid hydroperoxide glutathione peroxidase against membrane-damaging lipid peroxidation. In situ reduction of phospholipid and cholesterol hydroperoxides[J]. The Journal of Biological Chemistry,1990,265(1):454-461. [7]祝璟琳,王国良. 鱼类HSP70的研究进展[J]. 宁波大学学报:理工版,2007,20(4):446-450. [8]Mambula S S, Calderwood S K. Heat shock protein 70 is secreted from tumor cells by a nonclassical pathway involving lysosomal endosomes[J]. The Journal of Immunology,2006,177(11):7849-7857. [9]Yar Ahmadi P Y, Farahmand H, Kolangi Miandare H, et al. The effects of dietary immunogen on innate immune response, immune related genes expression and disease resistance of rainbow trout (Oncorhynchus mykiss) [J]. Fish & Shellfish Immunology,2014,37(2):209-214. [10]Wang X, Kim K, Bai S C, et al. Effects of the different levels of dietary vitamin C on growth and tissue ascorbic acid changes in parrot fish (Oplegnathus fasciatus)[J]. Aquaculture,2003,215(1):203-211. [11]Chen R, Lochmann R, Goodwin A E, et al. Effects of dietary vitamins C and E on alternative complement activity, hematology, tissue composition, vitamin concentrations and response to heat stress in juvenile golden shiner (Notemigonus crysoleucas)[J]. Aquaculture,2004,242(1/4):553-569. [12]高明辉. VC、VE对亚硝酸盐胁迫下异育银鲫血液指标及抗氧化能力的影响[D]. 武汉:华中农业大学,2008. [13]刘扬,池磊,冯琳,等. 维生素C对建鲤肠上皮细胞氧化损伤保护作用的研究[J]. 动物营养学报,2012,24(8):1503-1511. [14]武迪,管婷婷,张思雨,等.维生素C对泥鳅幼鱼生长和免疫的影响[J].水产科学,2016,35(6):658-662. [15]熊铧龙,姚俊杰,安苗,等.普安银鲫胚胎发育中抗氧化酶活性及外源维生素C的作用[J].水产科学,2014,33(10):631-634. [16]明建华,谢骏,徐跑,等. 大黄素、维生素C及其配伍对团头鲂生长、生理生化指标、抗病原感染以及两种HSP70s mRNA表达的影响[J]. 水产学报,2010,34(9):1447-1459. [17]万金娟. 维生素C对团头鲂幼鱼生长及免疫的影响[D]. 南京:南京农业大学,2012. [18]秦国兵,熊伟,姚俊杰,等. 维生素C对亚硝酸盐胁迫下匙吻鲟仔鱼非特异性免疫及抗氧化能力的影响[J]. 河南农业科学,2018,47(7):132-136. [19]Otte J C, Andersson C, Abrahamson A, et al. A bioassay approach to determine the dioxin-like activity in sediment extracts from the Danube River ethoxyresorufin-O-deethylase induction in gill filaments and liver of three-spined sticklebacks (Gasterosteus aculeatus L.)[J]. Environment International,2008,34(8):1176-1184. [20]Santos M A, Pacheco M, Ahmad I. Anguilla anguilla L. antioxidants responses to in situ bleached kraft pulp mill effluent outlet exposure[J]. Environment International,2004,30(3):301-308. [21]单书宝,浦佩玉. 氧自由基与临床 [J]. 国外医学:神经病学神经外科学分册,1995,22(2):94-96. [22]方允中,杨胜,伍国耀. 自由基稳衡性动态生理科学进展[J]. 生理科学进展,2004,35(3):199-204. [23]李勇男. 水中添加维生素C缓解鲤鱼运输应激的研究[D]. 无锡:江南大学,2016. [24]明建华. 大黄素和维生素C对团头鲂生长、非特异性免疫以及抗应激的影响[D]. 南京:南京农业大学,2011. [25]周传朋,刘波,谢骏,等. 抗应激制剂对运输团头鲂血清生化和肝脏抗氧化指标的影响[J]. 水产科学,2014,33(1):8-14. [26]姜亚军,家声,周君富.脑出血患者氧化与抗氧化状态的研究[J].铁道医学,1997,25(6):335-338. [27]Nemcsók J, Rakonczay Z, Kása P, et al. Effects of methidathion on distribution of molecular forms of acetylcholinesterase in carp, as revealed by density gradient centrifugation[J]. Pesticide Biochemistry and Physiology,1990,37(2):140-144. [28]Moore A, Waring C P. Sublethal effects of the pesticide Diazinon on olfactory function in mature male Atlantic salmon parr[J]. Journal of Fish Biology,1996,48(4):758-775. [29]赵丹. 急、慢性缺氧对斑马鱼幼鱼神经系统损伤机制的初步研究[D]. 广州:南方医科大学,2014. [30]齐红莉,张树林,戴伟. 农药对水产动物毒理的研究现状[J]. 水利渔业,2005,25(1):73-75. [31]谢芳,周志俊,吴强恩,等. 乐果对U87细胞HSP70表达及AChE活性的影响[J]. 环境与职业医学,2005,22(2):105-108. [32]杨东仁,李秋生,牛勇,等. 有机磷农药对骨骼肌细胞C-FOS以及HSP70表达的影响[J]. 中华劳动卫生职业病杂志,1999,17(3):172-173. [33]郭雅洁,梁沛,高希武. 烯啶虫胺、毒死蜱和高效氯氰菊酯对烟粉虱地中海隐种hsp70mRNA的诱导表达[J]. 昆虫学报,2013,56(1):29-38.
2020, Vol. 39 
