当归(粗)多糖对麻黄素肝组织损伤的保护作用

doi:10.16098/j.issn.0529-1356.2015.03.016

摘要/Abstract

摘要：

目的探讨当归多糖对麻黄素致肝损伤的保护作用。方法 72只小鼠分为正常对照组、实验对照组、当归多糖1组和当归多糖2组4个组，实验对照组和当归多糖组连续腹腔注射0.2ml浓度为4.0g/L麻黄素溶液15d(每天2次)，正常对照组注射等量生理盐水，当归多糖1组和2组在腹腔注射麻黄素溶液1h后，灌胃0.3ml(12.5g/L、25.0g/L)当归（粗）多糖溶液，正常对照组和实验对照组灌胃等量的生理盐水。分别于5、10、15d用光学显微镜技术观察各组小鼠肝组织结构的变化，用比色法检测小鼠血浆γ-谷氨酰转移酶(γ-GT)、碱性磷酸酶(AKP)、谷丙转氨酶(ALT)、谷草转氨酶(AST)及肝组织超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量的变化，用免疫组织化学方法检测小鼠肝组织Bax蛋白、Csapase-3蛋白表达的变化。结果实验对照组小鼠肝组织有不同程度的损伤，肝细胞索排列紊乱，肝细胞疏松，肝血窦扩张，血浆γ-GT、AKP、ALT、AST活性明显升高，肝组织SOD活性显著降低，MDA含量显著升高，Bax、Caspase-3蛋白的表达显著升高；当归多糖组小鼠肝组织病理损伤明显减轻，血浆γ-GT、AKP、ALT、AST活性明显降低，肝组织SOD活性明显升高，MDA含量显著降低，Bax、Caspase-3蛋白的表达有所降低。结论当归（粗）多糖可提高肝组织细胞的抗氧化能力，抑制细胞凋亡，促进组织细胞损伤修复，对麻黄素致肝损伤具有一定的保护作用。

Abstract:

Objective To investigate the protection of Angelica sinensis polysaccharide(ASP) on ephedrine induced hepatic injury in mice. Methods Seventy-two mice were randomly divided into 4 groups：the natural control group，experiment control group，ASP1 group， and ASP2 group.The experimental control group and ASP groups were intraperitoneally injected with a 4.0g/L of ephedrine 0.2ml twice a day for 15 days， and the natural control group was injected with a same amount of saline.The ASP groups were irrigated to stomach with 0.3ml Angelica sinensispolysaccharide solution (12.5g/L，25.0g/L)1 hour after ephedrine injection，the natural control group and experiment control group were irrigated to stomach with a same amount of saline.The mice were sacrificed at 5 day，10day，and 15 day after treatment，respectively.The morphological observation was used to observe pathological changes of hepatic tissues.Activities of hepatic histologicalsuperoxide dismutase（SOD）and plasma γ-glutamyltransferase(γ-GT), alkaline phosphatase (AKP), glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminase(AST)，as well as content of malonaldehyde（MDA）were detected by colorimetry.The expressions of Bax protein and Caspase-3 protein were measured by immunohistochemistry in liver of mice. Results There were various degree damages in hepatic tissues，hepatic cords were disorganized，liver cells were loose and liver sinusoid expanded，the activity of γ-GT、AKP、ALT、AST in plasma were significantly increased，the activity of SOD in liver tissue was significantly reduced，the content of MDA in liver tissue was increased significantly，the expression of Bax and Caspase-3 protein in liver were significantly increased.Compared with the experimental control group，pathological damage of liver was significantly improved，the activity of plasma γ-GT, AKP, ALT, AST were significantly reduced，the activity of SOD in liver tissue was increased significantly，however the content of MDA was significantly decreased and the expressions of Bax and Caspase-3 protein in liver were reduced in Angelica sinensis polysaccharide group. ConclusionAngelica sinensis polysaccharide can improve liver tissue antioxidant capacity and promote reparation of cell injury and protect hepatic injury which is induced by ephedrine.

孟茹李重阳俞诗源* 袁静魏娉婷. 当归(粗)多糖对麻黄素肝组织损伤的保护作用[J]. 解剖学报, 2015, (3): 379-386.

MENG Ru LI Chong-yang YU Shi-yuan* YUAN Jing WEI Ping-ting. Protection of Angelica sinensis polysaccharide on liver histological structure in mice injured by ephedrine[J]. AAS, 2015, (3): 379-386.

参考文献

［1］Pan ZhH，Sun XR.The safety evaluation of ephedra and its preparation aboard［J］.Chinese Journal of Pharmacovigilance，2007，4(2)：111-116.(in Chinese)
潘周华，孙晓如.国外对麻黄及其制剂的安全性评价［J］.中国药物警戒，2007，4(2)：111-116.
［2］Shekelle P G，Hardy ML, Morton SC，et al.Efficacy and safety of ephedra and ephedrine for weight loss and athletic performance: a meta\|analysis［J］. JAMA，2003，289(12)：1537-1545.
［3］Li ChY，Wang JJ，Yu ShY.Effect of Chinese traditional medicine on activities of antioxidant enzyme and special protein of kidney in ephedrine addiction mice［J］.Acta Anatomica Sinica，2013，3(44)：372-378.(in Chinese)
李重阳，王锦锦，俞诗源.中药复方对麻黄素成瘾小鼠肾抗氧化酶和特异蛋白的影响［J］.解剖学报，2013，3(44)：372-378.
［4］Nadir A，Agrawal S，King PD, et al.Acute hepatitis associated with the use of a Chinese herbal product，ma-huang［J］. Am J Gastroenterol，1996，91(7)：1436-1438.
［5］Sun J，Li ChY，Yu ShY，et al.Effects of ephedrine on structure and polypeptides in the liver of filial mice［J］.Acta Anatomica Sinica，2012，43(3)：398-404.(in Chinese)
孙隽，李重阳，俞诗源，等.麻黄素对仔鼠肝组织结构和相关活性物质的影响［J］.解剖学报，2012，43(3)：398-404.
［6］Yu F.Protective effects of Angelica sinensis polysaccharide on immunological liver in jury induce by concanavalin A in mice［J］.Journal of Chinese Medicinal Materials，2009，32(10)：1580-1583.(in Chinese)
俞发.当归多糖对刀豆蛋白A所致小鼠免疫性肝损伤的保护作用［J］.中药材，2009，32(10)：1580-1583.
［7］Cantner F，Leist M，Lohse AW，et al.Concanavalin A induced T-cell-mediated hepatic injury in mice：the role of tumormecrosis factor［J］.Hepatology，1995，21(1)：190-198.
［8］Sun YL，Yang PF，Wu HX，et al.Optimization of extraction technology for polysaccharide from angelica sinensis［J］.Journal of Chinese Institute of Food Science and Technology，2009，5(9)：130-134.(in Chinese)
孙元琳，杨萍芳，吴海霞, 等.水法提取当归多糖工艺条件优化［J］.中国食品学报，2009，5(9)：130-134.
［9］Zhang WJ. Sugar Review and Biochemical Research Techniques［M］.Hangzhou：Zhejiang University Press，1998:30-50.(in Chinese)
张惟杰，糖复核生化研究技术［M］.杭州：浙江大学出版社，1998:30-50.
［10］Feng HL，Yu ShY，Wang Y，et al.Histological structure and expression of KGF，C-Fos and Bax in the lung of Aguila chrysaetos［J］.Chinese Journal of Zoology 2009，44(1)：33-38. (in Chinese)
冯红丽，俞诗源，王昱，等.金雕肺的组织结构及KGF、c-Fos和Bax的表达［J］.动物学杂志，2009，44（1）：33-38.
［11］Jaeschke H, Gores GJ, Cederbaum AI, et al.Mechanisms of hepatotoxicity［J］.Toxicol Sci，2002，65(2)：166-176.
［12］Liu J，Peng RX，Le J，et al.Isolation，purification and partial properties of polysaccharides from Angelica Sinensis［J］.West China Journal of Pharmaceutical Sciences，2004，19(6):412-414.(in Chinese)
刘娟，彭仁琇，乐江，等.当归多糖的分离纯化及其部分理化性质的研究［J］.华西药学杂志，2004，19(6):412-414.
［13］Ning KJ，Zhang ZM，Si YY，et al.Electron microscope observation of radix Angelicae Sinensis on protection with the liver injury caused by carbon tetrachloride in mice［J］.Chinese Journal of Experimental Traditional Medical Formulae，2009，15(12)：68-70.(in Chinese)
宁康健，张子明，司阳洋，等.当归对四氯化碳肝损伤保护作用的电镜观察［J］.中国实验方剂学杂志，2009，15(12)：68-70.
［14］Yang TH，Jia M，Mei QB.Immunoloregulation effect of polysaccharide isolated from Angelica Sinensis on mouse［J］.Chinese Traditional Patent Medicine，2005，27(5):563-565.(in Chinese)
杨铁虹，贾敏，梅其炳.当归多糖对小鼠免疫功能的调节作用［J］.中成药，2005，27(5):563-565.
［15］Hu XP，Li YY，Li XH，et al.The advances of angelica polysaccharide composition and pharmacology［J］.Journal of Chinese Medicinal Materials，2004，27(1)：70-72.(in Chinese)
胡小平，李玉云，李先何，等.当归多糖的成分及药理学研究新进展［J］.中药材，2004，27(1)：70-72.
［16］Yuan DF，Wang HJ.The significance of serum ALT、γ-GT、CG、HA diagnosis in alcoholic liver disease ［J］.Journal of Radioimmunology，2005，2(18)：111-112.(in Chinese)
袁登峰，王海菊.血清ALT、γ-GT、CG、HA测定在酒精性肝病诊断中的意义［J］.放射性免疫学杂志，2005，2(18)：111-112.
［17］Li JT，Wang JL，Fu ShG.The activity changes of alkaline phosphatase and its ultrastructual localization during liver regeneration rats ［J］.Acta Anatomica Sinica，2004，4(35)：392-395.(in Chinese)
李金亭，王俊丽，傅山岗.大鼠肝再生过程中碱性磷酸酶活性变化及其超微细胞化学研究［J］.解剖学报，2004，4(35)：392-395.
［18］Lü JT，Tang H.Experimental study of the protective effect on liver damage by treatment with polysaccharide fructus ligustri［J］.Chinese Journal Of Hospital Pharmacy，2010，12(30):1024-1026.(in Chinese)
吕娟涛，汤浩.女贞子多糖对肝损伤保护作用的实验研究［J］.中国医院药学杂志，2010，12(30):1024-1026.
［19］Wang BL，Zhang YM，Hou YN，et al.Effects of cadmium and lead on transaminase activities in tissues of Misgurnus anguillicaudatus［J］.Journal of Lanzhou University（Natural Sciences），2006，42(3)：67-70.(in Chinese)
王丙莲，张迎梅，侯亚妮，等.镉铅对泥鳅组织转氨酶活性的影响［J］.兰州大学学报(自然科学版)，2006，42(3)：67-70.
［20］Cao G，Zhao J，Chen HG，et al.Effect of angelica sinensis polysaccharide on experimental hepatic injury in mice［J］.Journal of Xianning College，2004，18(5)：332.(in Chinese)
操刚，赵骥，陈红光，等.当归多糖对小鼠实验性肝损伤的影响［J］.咸宁学院学报，2004，18(5)：332.
［21］Liang R，Zhong Y.Free Radical Biology［M］.Beijing: Higher Education Press，2007:1-40.(in Chinese)
梁荣，中洋.自由基生物学［M］.北京：高等教育出版社，2007:1-40.
［22］Kikugawa K， Kosugi H.Detection and quantitative determination of lipid peroxidation in living bodies: thiobarbituric acids test ［J］.Japanese Journal of Toxicology And Environmental Health，1993，39(1)：1-19.
［23］Chen Y，Shen ShL，Chen HX.Analysis of ephedrine and its metabolites in rat urine by HPLC-ESI-ITMSn［J］.Acta Pharmaceutical Sinica，2005，40(9)：838-841.(in Chinese)
陈勇，沈少林，陈怀侠.HPLC-ESI-ITMSn法鉴定麻黄碱及其大鼠体内主要代谢产物［J］.药学学报，2005，40(9)：838-841.
［24］Benichou C.Criteria of drug-induced liver disorders.Report of an international consensus meeting［J］.Hepatology, 1990，11(2)：272-276.
［25］Huang YS，Chern HD，Su WJ，et al.Cytochrome P450 2E1 genotype and the susceptibility to antituberculosis druginduced hepatitis［J］.Hepatology，2003，37(4)：924-930.
［26］Xu X，Qu CQ.Mechanisms of drug-induced liver injury［J］.Medical Recapitulate，2008，5(14)：747-749.(in Chinese)
徐鑫，屈彩琴.药物性肝损伤机制［J］.医学综述，2008，5(14)：747-749.
［27］Li GR，Lü ChY，Yang ShY.Study on isolation of angelica sinensis polysaccharide and its effects on antiactive oxygen free radicals［J］.Journal of Nanhua University，2002，3(16)：18-20.(in Chinese)
李贵荣，吕昌银，杨胜圆.当归多糖清除活性氧自由基作用的研究［J］.南华大学学报，2002，3(16)：18-20.
［28］Liu L，Lin J.Overview of gastric mucosal healing and apoptosis［J］.Modern Journal of Integrated Traditional Chinese and Western Medicine，2008，17(14)：2267-2268.(in Chinese)
刘磊，林洁.胃黏膜愈合和细胞凋亡概述［J］.现代中西医结合杂志，2008，17(14)：2267-2268.
［29］Yu ShY，Liu XY，Wu ZhZ，et al.Protection of Chinese medicine com pound on ovarian histological structure in filial mice of ephedrine injury［J］.Journal of North West Normal University (Natural Science)，2014，50（3）：79-87.(in Chinese)
俞诗源，刘小媛，吴转子，等.中药复方液对麻黄素损伤小鼠卵巢的保护作用［J］.西北师范大学学报：自然科学版，2014，50（3）：79-87.
［30］Muzio M，Salvesen GS，Dixit VM.FLICE induced apoptosis in a cell-free system.Cleavage of caspase zymogens［J］. J Biol Chem，1997，272(5)：2952-2956.
［31］Ding H，Peng RX，Yu JP.The protection of Angelica sinensis polysaccharide on NOS and apoptosis related genes in mice liver by immunological liver injured［J］.Chinese Journal of Hepatology，2001，7(9)：50-52.(in Chinese)
丁虹，彭仁琇，于皆平.当归多糖对免疫性肝损伤小鼠肝中一氧化氮合酶及凋亡相关基因的影响［J］.中国肝脏病杂志，2001，7(9)：50-52.
［32］Zheng M，Wang YP.Experimental study on effect of Angelica polysaccharide in inhibitory proliferation and inducing differentiation of K652 cells［J］.Chinese Journal of Integrated Traditional and Western Medicine，2002，22 (1)：54-57.(in Chinese)
郑敏，王亚平.当归多糖对K562细胞增殖与诱导分化的研究［J］.中国中西医结合杂志，2002，22 (1)：54-57.
［33］Yang QH，Jia M，Mei QB.Effect of Angelica sinensis polysaccharide on lymphocyte proliferation and cytokine induction［J］.Journal of Chinese Medicinal Materials，2005，28(5)：405.(in Chinese)
杨钦红, 贾敏，梅其炳.当归多糖组分促进淋巴细胞增殖及对 IL-2和INF-γ的诱生作用［J］.中药材，2005，28(5)：405.