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枸杞多糖对四氯化碳诱导建鲤原代肝细胞损伤的保护作用研究
引用本文:刘英娟,杜金梁,贾睿,曹丽萍,王佳豪,殷国俊.枸杞多糖对四氯化碳诱导建鲤原代肝细胞损伤的保护作用研究[J].上海海洋大学学报,2014,23(5):718-725.
作者姓名:刘英娟  杜金梁  贾睿  曹丽萍  王佳豪  殷国俊
作者单位:南京农业大学无锡渔业学院,中国水产科学研究院淡水渔业研究中心,南京农业大学无锡渔业学院,中国水产科学研究院淡水渔业研究中心,南京农业大学无锡渔业学院,中国水产科学研究院淡水渔业研究中心
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(2013JBFM11,2013JBFM12) ;国家自然基金青年科学基金(31202002、31200918);江苏省自然科学基金(BK2012535)
摘    要:研究中药枸杞多糖(Lycium barbarum polysaccharide,LBP)对四氯化碳(CCl4)诱导的建鲤(Cyprinus carpiovar Jian)肝细胞急性损伤的保护作用。利用8 mmol/L四氯化碳构建建鲤肝细胞损伤模型,用不同浓度的枸杞多糖处理肝细胞,设空白对照组、枸杞多糖对照组(0.4 mg/mL)、模型组(CCl4)、预防组(CCl4处理前加枸杞多糖孵育,0.1、0.2和0.4 mg/mL)、治疗组(CCl4处理后加枸杞多糖孵育,0.1、0.2和0.4 mg/mL)和预防治疗组(枸杞多糖孵育后经CCl4处理再经枸杞多糖孵育,0.1、0.2和0.4 mg/mL)。12 h后,收集细胞和细胞培养液,然后测定谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、细胞活力、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、免疫球蛋白M(IgM)及药物代谢酶细胞色素P450 2E1(CYP2E1)等生化指标。研究结果表明:CCl4处理后可以显著增加GPT、GOT、LDH、MDA、TNF-α、IL-1β、IgM及CYP2E1的含量,降低SOD活性和细胞活力;枸杞多糖处理组可不同程度地抑制上述生化指标的变化,其中预防组及预防治疗组对于转氨酶(GPT、GOT)、LDH、MDA、细胞因子(TNF-α、IL-1β)、IgM及CYP2E1等活性的升高皆有显著抑制作用,同时显著提高SOD酶活性及细胞活力,且呈剂量依赖性;而治疗组仅对LDH、TNF-α及CYP2E1有显著的抑制作用。综上结果,认为枸杞多糖对四氯化碳致建鲤肝损伤具有一定保护作用,可以应用于鱼类肝损伤的防治。

关 键 词:枸杞多糖  急性肝损伤  四氯化碳  原代培养肝细胞  建鲤
收稿时间:3/5/2014 12:00:00 AM
修稿时间:2014/4/15 0:00:00

Hepatoprotective effects of Lycium barbarum polysaccharides on carbon tetrachloride induced primary hepatocyte injury in Jian Carp
LIU Ying-juan,DU Jin-liang,JIA Rui,CAO Li-ping,WANG Jia-hao and YIN Guo-jun.Hepatoprotective effects of Lycium barbarum polysaccharides on carbon tetrachloride induced primary hepatocyte injury in Jian Carp[J].Journal of Shanghai Ocean University,2014,23(5):718-725.
Authors:LIU Ying-juan  DU Jin-liang  JIA Rui  CAO Li-ping  WANG Jia-hao and YIN Guo-jun
Institution:Wuxi Fisheries College, Nanjing Agricultural University,Freshwater Fisheries Research center, Chinese Academy of Fishery Sciences,Wuxi Fisheries College, Nanjing Agricultural University,Freshwater Fisheries Research center, Chinese Academy of Fishery Sciences,Wuxi Fisheries College, Nanjing Agricultural University,Freshwater Fisheries Research center, Chinese Academy of Fishery Sciences
Abstract:In order to study the hepatoprotective effects of Chinese herbal medicine Lycium barbarum polysaccharide(LBP) against carbon tetrachloride-induced hepatocyte injury in Cyprinus carpio var. jian. Different concentrations of LBP (0.1 mg/mL, 0.2 mg/mL, 0.4 mg/mL) were added to the carp primary hepatocytes before(pre-treatment) ,after(post-treatment)and both before and after( pre-and post-treatment)incubation of the hepatocytes with CC14. After 12 h, the cell and supernatant were collected, and then the levels of glutamatepyruvate transaminase ( GPT ), glutamate oxalate transaminase ( GOT ), lactate dehydrogenase ( LDH), malondialdehyde ( MDA ), superoxide dismutase ( SOD ), cell viability, tumor necrosis factor-or ( TNF- α ), Interleukin-1 β ( IL-1β ), immunoglobulin M ( IgM ) and cytochrome p450 2El ( CYP2E1 ) were measured. The results showed that CC14 at 8 mmol/L in the culture medium significantly elevated the levels of GPT, GOT, LDH, MDA, TNF-α, IL-1β, IgM and CYP2E1, and significantly reduced levels of SOD and cell viability; however, treatment with LBP attenuated the adverse changes of these biochemical indexes, especially in pre-treatment and pre-and post-treatment; the changed levels of GPT, GOT, LDH, MDA,TNF- α, IL-16, IgM and CYP2Eland the elevated levels of SOD and cell viability were significantly inhibited in a dose-dependent manner, while the post-treatment of hepatocytes with LBP only significantly reduced the levels of LDH,TNF-α and CYP2E1. Based on the results, it is indicated that LBP may have the hepatoprotective effects against carbon tetrachloride-induced hepatocyte injury in Cyprinus carpio var. jian and may be used as a hepatoprotective agent in fish liver diseases.
Keywords:Lycium barbarum polysaccharide  Acute liver injury  Carbon tetrachloride  primary hepatocytes
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