[1]张春云,曹丽萍,杜金梁,等.Nrf2介导姜黄素对四氯化碳诱导鲫鱼肝损伤的保护作用[J].江苏农业学报,2018,(01):114-121.[doi:doi:10.3969/j.issn.1000-4440.2018.01.017]
　ZHANG Chun-yun,CAO Li-ping,DU Jin-liang,et al.Nrf2 mediate protective effects of curcumin against carbon tetrachloride(CCl4)-induced liver injury in Carassius auratus[J].,2018,(01):114-121.[doi:doi:10.3969/j.issn.1000-4440.2018.01.017]
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Nrf2介导姜黄素对四氯化碳诱导鲫鱼肝损伤的保护作用()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年01期
页码:
114-121
栏目:
畜牧兽医·水产养殖
出版日期:
2018-02-25

文章信息/Info

Title:
Nrf2 mediate protective effects of curcumin against carbon tetrachloride(CCl4)-induced liver injury in Carassius auratus
作者:
张春云1; 曹丽萍2; 3; 杜金梁2; 3; 贾睿2; 3; 顾郑琰1; 殷国俊1; 2; 3; Galina Jeney3; 4
（1.南京农业大学无锡渔业学院，江苏无锡214081；2.中国水产科学研究院淡水渔业研究中心，农业部淡水渔业和种质资源利用重点实验室，江苏无锡214081；3.中国水产科学研究院淡水渔业研究中心，农业部鱼类免疫药理学国际联合实验室，江苏无锡214081；4.Research Institute for Fisheries, Aquaculture and Irrigation, Anna light 8, Szarvas H-4440, Hungary）
Author(s):
ZHANG Chun-yun1; CAO Li-ping2; 3; DU Jin-liang2; 3; JIA Rui2; 3; GU Zheng-yan1; YIN Guo-jun1; 2; 3; GALINA Jeney3; 4
(1. Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China;2.Key Laboratory of Freshwater Fisheries and Germplasm Resource Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences
关键词:
核因子E2相关因子2(Nrf2 ); 姜黄素; 四氯化碳; 肝损伤; 鲫鱼
Keywords:
Nrf2; curcumin; CCl4; hepatic injury; Carassius auratus
分类号:
S965.117
DOI:
doi:10.3969/j.issn.1000-4440.2018.01.017
文献标志码:
A
摘要:
为了评价姜黄素对四氯化碳（CCl4）诱导鲫鱼肝损伤的保护作用以及核因子NF-E2相关因子(Nrf2)的调控作用，采用精密肝切片技术(Precision-cut liver slices, PCLS)分离肝组织并进行体外培养，以四氯化碳诱导建立急性肝损伤模型，同时用不同浓度(1 μg/ml、5 μg/ml和10 μg/ml)的姜黄素处理肝组织切片，收集上清液及肝组织。检测谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px )、过氧化氢酶(CAT)、碱性磷酸酶(AKP)活性，以及丙二醛(MDA)、谷胱甘肽(GSH)、总蛋白(TP)、白蛋白(Alb)含量和总抗氧化能力(T-AOC)等生化指标，同时检测Nrf2和Keap1 基因表达。结果表明：姜黄素可以改善CCl4诱导的肝损伤，其浓度为1 μg/ml和5 μg/ml时能有效抑制GPT、GOT、LDH和AKP活性的升高，提高SOD、GSH-Px活性及T-AOC和GSH含量，降低MDA的生成，上调Nrf2基因表达。可见，姜黄素对CCl4诱导的鲫鱼肝损伤有保护的作用，其作用机制可能是姜黄素能激活Nrf2基因在肝细胞中的表达，调节氧化应激，从而减轻CCl4对肝损伤。
Abstract:
To evaluate the protective effects of curcumin on liver injury induced by carbon tetrachloride (CCl4) and the regulatory role of nuclear factor NF-E2 related factor (Nrf2) in Carassius auratus, the liver tissue was isolated by precision-cut liver slices (PCLS) and cultured in vitro. The PCLS was treated with carbon tetrachlorideand curcumin (1 μg/ml, 5 μg/ml and 10 μg/ml). Then the culture supernatants and slices were collected for the analyses of biochemical parameters, such as activities of glutamatepyruvate transaminase (GPT), glutamate oxalatetransaminase (GOT), lactate dehydrogenase (LDH), superoxide dismutase (SOD), glutathione peroxidase(GSH-Px), catalase (CAT) and alkaline phosphatase(AKP), and contents of malondialdehyde (MDA), glutathione (GSH), total protein (TP), albumin (Alb), and total antioxidant capacity (T-AOC) and gene expression of Nrf2 and Keap1. The results showed that curcumin could inhibited the liver damage caused by CCl4. Curcumin at 1 μg/ml and 5 μg/ml significantly reduced the levels of GPT, GOT, LDH and AKP, increased the activities of SOD and GSH-Px, T-AOC and GSH content, suppressed the production of MDA, and up-regulated the expression of Nrf2 in the CCl4-induced liver. In conclusion, curcumin had protective effect on liver injury induced by CCl4. The hepatoprotective action of curcumin was probably accomplished by regulating Nrf2 expression to elevate cellular antioxidative capacity and inhibit lipid peroxidation.

参考文献/References:

[1]OTTU O J, ATAWODI S E, ONYIKE E. Antioxidant hepatoprotective and hypolipidemic effecs of methanolic root extract of Cassia singueana in rats following acute and chronic carbon tetrachloride intoxication[J]. Asian pacific Journal of Tropical Medicine, 2013, 2: 609-615.
[2]黄燕平,杨先乐,湛嘉,等. 水产动物疾病控制的研究和进展[J]. 上海水产大学学报, 2004, 13(1): 60-66.
[3]CABELLO F C. Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal health and for the environment[J]. Environmental Microbiology, 2006, 8(7): 1137-1344.
[4]贾睿,曹丽萍,杜金梁,等. 水飞素对四氯化碳致鲫鱼肝(细胞)损伤的保护和抗氧化作用[J]. 中国水产科学, 2012, 3: 551-560.
[5]苏岭,刘红柏,王荻,等. 四种复方中药和黄芪多糖对鲫鱼生长、组织中NO含量与NOS活性的影响[J]. 水产学杂志, 2010, (23)3: 11-15.
[6]GALINA J, YIN G, ARD L, et al. The use of immunostimulating herbs in fish. An overview of research[J]. Fish Physiology and Biochemistry, 2009, 35(4): 669-676.
[7]LIU X, WANG D, LU T Y. The effects of compound Chinese medicine immune additives on growth performance and antioxidant capacity of Acipenser schrenckii[J]. Animal Husbandry and Feed Science, 2012, 4(5): 221-225.
[8]李霞,宋其林,陈炳卿. 姜黄素抗诱变的研究[J]. 卫生研究, 1998, 27(4): 145-147.
[9]石晶,顾军,邓心新,等. 姜黄素对大鼠心肌缺血性损伤的保护作用[J]. 中国药理学通报, 1998, 14(2): 145-147.
[10]石晶,王中孝,田亚平,等. 姜黄素对高脂血症大鼠血浆和肝脏超氧化物歧化酶和脂质过氧化物的影响[J]. 中草药, 1997, 28(5): 285-287.
[11]汪海慧,成扬. 姜黄素药理作用的研究进展[J]. 上海中医药大学学报, 2007, 21(6): 73-76.
[12]胡忠泽,杨久峰,谭志静,等. 姜黄素对草鱼生长和肠道酶活力的影响[J]. 粮食与饲料工业, 2003(11): 29-30.
[13]FRYER J L, LANNAN C N. Three decades of fish cell culture: A current listing of cell lines derived from fishes[J]. Journal of Tissue Culture Methods, 1994, 16: 87-94.
[14]HONG Y, SCHARTL M. Establishment and growth responses of early medakafish (Oryzias latipes) embryonic cells in feeder layer-free cultures[J]. Molecular Marine Biology & Biotechnology, 1996, 5(5): 93-104.
[15]KELLY R K, MILLER H R, NIELSON O, et al. Fish cell culture: characteristics of a continuous fibroblastic cell line from walleye (Stlzosterion vltrem vireum) [J]. Fish Aqunt Science, 1980, 37: 1070-1075.
[16]刘永刚,赵进军. 姜黄素对四氯化碳损伤原代培养大鼠肝细胞的影响[J]. 中成药, 2003, 25(3): 222-224.
[17]FAROMBI E O, SHROTRIYA S, NA H K, et al. Curcumin attenuates dimethylnitrosamine-induced liver injury in rats through Nrf2-mediated induction of heme oxygenase-1[J]. Food & Chemical Toxicology, 2008, 46(4): 1279-1287.
[18]BALOGAN E, HEQUE M, GONG P, et al. Curcumin activatrs the haem oxygenane-I gene via regulation of Nrf2 and the antioxidant-responsive element[J]. Biochemical Journal, 2003,371(3): 887-895.
[19]LEE H S, LI L, KIM H K, et al. The protective effects of Curcuma longa Linn. Extract on carbon tetrachloride-induced hepatotoxicity in rats via upregulation of Nrf2[J]. Journal Microbiol Biotechnol, 2010, 20(9): 1331-1338.
[20]MALEKINEJAD H, ALIZADEH A, CHERAGHI H, et al. The protective effect of liquorice plant extract on CCl4-induced hepatotoxicity in common carp (Cyprinus carpio)[J]. Veterinary Research Forum, 2010, 1(3): 158-165.
[21]HAN J, GAO C, YANG S, et al. Betanin attenuates carbon tetrachloride (CCl4)-induced liver injury in common carp ( Cyprinus carpio L.)[J]. Fish Physiology & Biochemistry, 2014, 40(3): 865-874.
[22]曹丽萍,贾睿,丁炜东,等. 建鲤急性肝损伤模型的建立及当归提取物的保肝和抗氧化作用研究[J]. 大连海洋大学学报,2012,27(6): 551-557.
[23]ZHOU J F,CAO D,ZHU Y G,et al. A study on relationship of nitric oxide,oxidation,peroxidation,lipoperoxidation with chrinic cholecystities[J]. World Journal Gastroente ROL,2000,6: 501-507.
[24]杨吉文,谭小艳,茆永琴,等. 组织细胞内ATP的动态变化研究[J]. 现代生物医学进展,2009,9(21): 4142-4145.
[25]高见,董凌月,安威,等. GFER抑制四氯化碳对eHepG2细胞的损伤[J]. 中国组织化学与细胞化学杂志,2015,24(5): 447-451.
[26]BASU S. Carbon tetrachlotide-induced lipid peroxidation: eicosanoid formation and their regulation by antioxidant nutrients[J]. Toxicology,2003,189: 113-127.
[27]BERRY M N,HALLS H J,GRIVELL M B,et al. Techniques for pharmacological and toxicological studies with isolated hepatocyte suspensions[J]. Life Sciences,1992,51(1): 1-16.
[28]ROMERTO F J,BOSCH-MORELL F,ROMERO M J,et al. Lipid peroxidation products and antioxidants in human disease[J]. Environmental Health Perspectives,1998,106(Suppl5): 1229-1234.
[29]NAIK R S,MUJUMDAR A M,GHASKADBI S,et al. Protection of liver cells from ethanol cytotoxicity by curcumin in liver slice culture in vitro[J]. Ethnopharmaacol,2004,95(1): 31-37.
[30]SMUCKLER E A,BENDITT E P. Studies on carbon tetrachloride intoxication. Iii A subcellular defect in protein synthesis[J]. Biochemistry,1965,4(4): 671-679.
[31]FOLMAR L C,BONOMELLI S,MOODY T,et al. The effect of short-term exposure to three chemicals on the blood chemistry of the pinfish (Lagodon rhomboides)[J]. Archives of Environmental Contamination and Toxicology,1993,24(1): 83-86.
[32]宁康健,丁建华,代守志,等. 黄芩苷对小鼠四氯化碳肝损伤保护的研究[J]. 畜牧与饲料科学,2009,30(1): 4-5.
[33]熊文静. 姜黄素的醇提工艺及其对动物的肝保护研究[D]. 武汉:华中农业大学,2016.
[34]NAYAK S,SASHIDHAR R B. Metabolic intervention of aflatoxin B1 toxicity by curcumin[J]. Ethnopharmacol,2010,127(3): 641-652.
[35]荣爽,李珂,宋方方,等. 姜黄素对乙醇诱导的大鼠原代细胞损伤的防护作用[J]. 环境与健康杂志,2009,26(6): 487-489.
[36]邹国良. 姜黄素对CCl4所致急性肝损伤的保护研究[J]. 中国中医药现代远程教育,2010,8(12): 229-230.
[37]钟越,钟秀宏,孙燕美,等. 姜黄素拮抗四氯化碳致肝纤维作用[J]. 中国公共卫生,2015,31(4): 447-449.
[38]E1-AGAMY D S. Comparative effects of curcumin and resveratrol on aflatoxin B(1)-induced liver injury in rats[J]. Archives of Toxicology,2010,84(5): 389-396.
[39]MAGDA K,EZZ,GERMINE M HMDY,et al. The synergistic hepatoprotective effect of curcumin and ginger against carbon tetrachloride induced-liver fibrosis in rats[J]. Australian Journal of Basic and Applied Sciences,2011,5(9): 1962-1971.
[40]ELISABETH B,MARTHA H,PENGFEI G,et al. Curcumin activates the haem oxygenase-1 gene via regulation of Nrf2 and the antioxidant-responsive element[J]. Biochemical,2003,1371(3): 887-895.
[41]戈孟雪,屈强,周宏灏,等. 调控Nrf2-Keap1-ARE信号通路的中药有效成分研究进展[J]. 中国临床药理学与治疗学,2015,20(6): 707-712.
[42]ESMAEILI M A,ALILOU M. Naringenin attenuates CCl4-induced hepatic inflammation by the activation of an Nrf2-mediated pathway in rats[J]. Clinical & Experimental Pharmacology & Physiology,2014,41(6): 416-422.
[43]WU T,LI J,LI Y,et al. Antioxidant and hepatoprotective effect of swertiamarin on carbon tetrachloride-induced hepatotoxicity via the Nrf2/HO-1 pathway[J]. Cellular Physiology & Biochemistry,2017,41(6): 22-42.
[44]CHEN B,LU Y,CHEN Y,et al. The role of Nrf2 in oxidative stress-induced endothelial injuries[J]. J Endocrinol,2015,225(3): 83-99.
[45]QUE L L,WANG H X,CAO B S,et al. The regulation and functions of transcription factor Nrf2 in cancer chemoprevention and chemoresistance[J]. Journal of Chinese Pharmaceutical Sciences,2011,20(1): 5-19.
[46]WU S C,YUAN Y,HUI T,et al. Carthamus red from Carthamus tinctorius L. exerts antioxidant and hepatoprotective effect against CCl4-induced liver damage in rats via the Nrf2 pathway[J]. Journal of Ethnopharmacology,2013,148: 570-578.
[47]宋永周,关键,李明,等. Nrf2介导姜黄素对软骨细胞氧化应激损伤的保护作用[J]. 中国组织工程研究，2015,51(19): 8218-8222.
[48]KEUM Y S,CHOI B Y. Molecular and chemical regulation of the Keap1-Nrf2 signaling pathway[J]. Molecules,2014,19(7): 10074-10089.

备注/Memo

备注/Memo:
收稿日期：2017-10-12 基金项目：中央级公益性科研院所基本科研业务费专项资金项目(2017JBFZ01)；国家自然科学基金项目（31702318）；江苏省自然科学基金项目（BK20170218） 作者简介：张春云(1989-), 女，安徽宿州人，硕士，研究方向为鱼类免疫药理学。(E-mail)shanyun0729@163.com 通讯作者：殷国俊，(E-mail)yingj@ffrc.cn
更新日期/Last Update: 2018-03-06