高山红景天多酚对小鼠肝损伤的保护作用
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摘要
目的研究高山红景天多酚对小鼠肝损伤的作用。方法健康昆明种雄性小鼠随机分成6组:1)正常对照组;2)模型组;3)高山红景天多酚低剂量组;4)高山红景天多酚中剂量组;5)高山红景天多酚高剂量组;6)水飞蓟素阳性对照组。小鼠急性肝损伤模型分别通过腹腔注射D-氨基半乳糖(D-galactosamine,D-GalN)700 mg/kg联用脂多糖(Lipopolysaccharide,LPS)10μg/kg,腹腔注射对乙酰氨基酚(Acetaminophen,APAP)300 mg/kg,灌胃他克林(Tacrine,THA)35 mg/kg来分别制备。给药组在造模前第24h,12h,1h(2h)时分别灌胃高山红景天多酚(50,100,150 mg/kg),水飞蓟素(100 mg/kg),其他组给予相同体积的生理盐水。小鼠的肝损伤主要由血清指标丙氨酸氨基转化酶(ALT)、天冬氨酸氨基转化酶(AST)、肿瘤坏死因子(TNF-α)和组织中丙二醛(MDA)、谷胱甘肽(GSH)等水平来进行评价。并且用组织病理学和免疫组织化学方法观察每组肝组织病理变化及caspase-3和缺氧诱导因子(HIF-1α)的表达。结果D-GalN/LPS,APAP,THA可以诱导小鼠肝损伤,主要表现在小鼠血清ALT、AST水平的显著升高和肝组织MDA水平和GSH消耗的显著升高。并且肝组织发生严重的病理变化,也表达caspase-3和缺氧诱导因子-1α(HIF-1α)。提前给予高山红景天多酚不但可以减轻由D-GalN/LPS,APAP,THA诱导的肝组织病变程度和caspase-3和HIF-1α的表达,还显著降低小鼠血清中的ALT、AST水平,组织中的MDA水平和GSH的消耗。结论高山红景天多酚对D-GalN/LPS,APAP,THA诱导的小鼠肝损伤具有保护作用,其作用其机制可能是与清除自由基,抗氧化,抑制caspase-3和HIF-1α表达有关。
Objective To investigate the effect of Rhodiola sachalinensis polyphenol (RSP) on liver injure induced by D-galactosamine(D-GalN)/Lipopolysaccharide(LPS), Acetaminophen(APAP),Tacrine(THA) in mice.Methods Healthy male Kunming mice were randomly divided into 6 groups:Normal group,Model group,RSP groups(50,100,150 mg/kg,respectively),Silymarin groups(100 mg/kg).Mice were pretreated with RSP or Silymarin by orally,at 24h,12h and 1h(2h) before D-GalN/LPS,APAP,THA injection and sacrificed after 8h or 12h or 6h treated with D-GalN/LPS,APAP,THA.The liver accessed biochemically and histologically.To observe serum alanine aminotransferase(ALT),aspartate amino transferase(AST) and hepatic malondialdehyde(MDA),glutathione(GSH) et al.Furthermore, histopathological and immunohistochemical were performed after hematoxylin & esosin (H&E) staining and caspase-3,hypoxia-inducible transcription factor-1α(HIF-1α) staining.Results D-GalN/LPS,APAP,THA could induce liver injury by markedly increase serum ALT,AST,the haptic MDA and GSH depletion.Also significantly increase the histopathology change and caspase-3 or HIF-1αexpression in mice liver. Pretreated with RSP markedly reduced the leather liver injury and the expression of caspase-3 or HIF-1αin the experimental animals and significantly decrease ALT,AST and the haptic MDA.Furthermore RSP pretreatment also significantly alleviated the hepatic GSH.Conclusion These results suggest that RSP have protective effect on D-GalN/LPS,APAP,THA induced liver injury in mouse model,the potential mechanism was related on remove free radicals,antioxidation and inhibit the caspase-3, HIF-1αexpression.
Objective To investigate the effect of Rhodiola sachalinensis polyphenol (RSP) on liver injure induced by D-galactosamine(D-GalN)/Lipopolysaccharide(LPS), Acetaminophen(APAP),Tacrine(THA) in mice.Methods Healthy male Kunming mice were randomly divided into 6 groups:Normal group,Model group,RSP groups(50,100,150 mg/kg,respectively),Silymarin groups(100 mg/kg).Mice were pretreated with RSP or Silymarin by orally,at 24h,12h and 1h(2h) before D-GalN/LPS,APAP,THA injection and sacrificed after 8h or 12h or 6h treated with D-GalN/LPS,APAP,THA.The liver accessed biochemically and histologically.To observe serum alanine aminotransferase(ALT),aspartate amino transferase(AST) and hepatic malondialdehyde(MDA),glutathione(GSH) et al.Furthermore, histopathological and immunohistochemical were performed after hematoxylin & esosin (H&E) staining and caspase-3,hypoxia-inducible transcription factor-1α(HIF-1α) staining.Results D-GalN/LPS,APAP,THA could induce liver injury by markedly increase serum ALT,AST,the haptic MDA and GSH depletion.Also significantly increase the histopathology change and caspase-3 or HIF-1αexpression in mice liver. Pretreated with RSP markedly reduced the leather liver injury and the expression of caspase-3 or HIF-1αin the experimental animals and significantly decrease ALT,AST and the haptic MDA.Furthermore RSP pretreatment also significantly alleviated the hepatic GSH.Conclusion These results suggest that RSP have protective effect on D-GalN/LPS,APAP,THA induced liver injury in mouse model,the potential mechanism was related on remove free radicals,antioxidation and inhibit the caspase-3, HIF-1αexpression.
引文
[1]Asha,V.V.,Pushpangadan,P.,Preliminary evaluation of the anti-hepatotoxic activity of Phyllanthus kozhikodianus,Phyllanthus maderspatensis and Solanum indicum.Fitotherapia,1998,59:255-259
[2]Subramoniam,A.,Too many hepatoprotective drugs and a little hepatoprotection.Current Science,1995,69,898-899
[3]王亮,游冬青,陈杞.红景天药理研究进展.时珍国医国药,1999,10(4):296-296
[4]高学敏主编.中药学.第1版.北京:中国中医药出版社,2002:511
[5]高秋娜.红景天的药理与临床研究概述.河南中医,2001,22(3):76-77
[6]Min-Won Lee,Yeon-Ah Lee,Hyang-Mi Park,Sang-Hak Toh,Eun-Joo Lee,Hae-Dong Jang,Young-Ho Kim.Antioxidative phenolic compounds from the Roots of Rhodiola sachlinensis A.Bor.Medicinal Chemistry & Natural Products,2000,23(5):455-458
[7]Lei Chen,Hong-Yu Zhang.Cancer Preventive Mechanisms of the Green Tea Polyphenol(-)-Epigallocatechin-3-gallate.Molecules,2007,12:946-9
[8]Nan JX,Jiang YZ,Park EJ,Ko G,Kim YC,Sohn DH.Protecive effect of Rhodiola sachalinensis extract on carbon tetrachloride-induced liver injury in rats.Journal of Ethno-Pharmacology,2003,84:143-148
[9]Song EK,Kim JH,Kim JS,Cho H,Nan JX,Sohn DH,Ko GI,Oh H,Kim YC.Hepatoprotective phenolic constituents of Rhodiola sachalinensis on tacrine-induced cytotoxicity in Hep G2 cells.Phytother Res.,2003,17:563-565
[10]Williams,R.Classification,etiology,and considerations of outcome in acute liver failure.Semin.Liver Dis,1996;16:343-348
[11]Ostapowicz,G.,and W.M.Lee.Acute hepatic failure:a Western perspective.J Gastroenterol.Hepatol,2000;15:480-488
[12]Galanos,C.,Freudenberg,M.A.,Feutter,W..Galactosamine-induced sensitization to the lethal effects of endotoxin.Proc.Natl.Acad.Sci. U.S.A,1979;76:5939-5943
[13]WANG H,XU DX,Lu JW,Zhao L,Zhang C,Wei W.N-Acetyl cysteine attenuates lipopolysaccharide-Induced apoptotic liver damage in D-galactosamine-sensitized mice.Acta Pharmacol Sin,2007;28(11):1803-1809
[14]Tetsuko Fulkuda,Akira Mogami,Hideki Tannaka,Tsutomu Yoshikawa,Masao Hisadome,Hirotsugu Komatsu.Y-40138,amultiple cytokine production modulator,protects against D-galactosamine and lipopolysaccharide-induced hepatitis.Life Sciences,2006;79:822-827
[15]Anne M.Larson,MD,FACP,AGAF.Acetaminophen Hepatotoxicity.Clin Liver Dis,2007,11:525-548
[16]O Brien,J.T.,Eagger,S.,Levy,R..Effects of tetrahydroaminoacridine on liver function in patients with Alzheimer's disease.Age Ageing,1991,20:129-131
[17]Arvelo,M.B.,Cooper,J.T.,Longo,C.,Daniel,S.,Grey,S.T.,Mahiou,J.,Czismadia,E.,Abu-Jawdeh,G.,Ferran,C..A20 protects mice from D-galactosamine/lipopolysaccharide acute toxic lethal hepatitis.Hepatology,2002,35:535-43
[18]Huiping Wang,Yan Li.Protective effect of bcyclol on acute hepatic failure induced by lipopolysaccharide and D-galactosamine in mice.European Journal of Pharmacology,2006,534:194-201
[19]雷桂兰,吴中华,刘世旺,李树文.苦荆茶中多酚类物质的含量测定.长江大学学报,2005,1(2):37-39
[20]Sung-Hwa Kim1,Yeong Shik Kim,Sam Sik Kang,KiHwan Bae,Tran Manh Hung,and Sun-Mee Lee1.Anti-apoptotic and Hepatoprotective Effects of Gomisin A on Fulminant Hepatic Failure Induced by D-Galactosamine and Lipopolysaccharide in Mice.J Pharmacol Sci,2008,106:225-233
[21]Brouckaert P,Fiers W.Tumor necrosis factor and the systemic inflammatory response syndrome.Curr Top Microbiol Immunol,1996;216:167-187
[22]田静.水飞蓟素抗肝损药理及其复合物的研究进展.海峡药 学,2004,16(5):7-9
[23] Mary Lynn Bajt, Anwar Farhood. Mitochondarlal Bax Translocation Accelerats DNA Fragmentation and cell necosis in murine model of acetaminophen hepatototoxicity. American Society for Pharmacology and Experimental Therapeutics, 2007,10:1-25
[24] Jain A, Soni M, Deb L, Jain A, Rout SP, Gupta VB, Krishna KL. Antioxidant and Hepatoprotective activity of ethanolic and aqueous extracts of Momordica dioica Roxb. Leaves. Journal of Ethno-Pharmacology, 2008,115:61-66
[25] Knight TR, Kurtz A, Bajt ML, Hinson JA, Jaeschke H. Vascular and hepatocellular peroxynitrite formation during acetaminophen toxicity: role of mitochondrial oxidant stress. Toxicol Sci, 2001, 62:212-220
[26] Laura P. James, Brian Donahower, Angela S., Burke, Sandra McCullough, Jack A. Hinson. Induction of the nuclear factor HJP-1a in acetaminophen toxicity: Evidence for oxidative stress. Biochemical and Biophysical Research Communications, 2006,3:171-176
[27] Stachlewitz RF, Arteel GE, Raleigh JA, Connor HD, Mason RP, Thurman RG. Development and Characterization of a New Model of Tacrine-Induced Hepatotoxicity: Role of the Sympathetic Nervous System and Hypoxia-Reoxygenation. The Journal of Pharmacology and Experimental, Therapeutics, 1997, 282(3): 1591-1599
[1]陈纪军等.德钦红景天的化学成分.云南植物研究,1999:21(4):525
[2]田军等.红景天挥发油的化学成分研究.军事医学科学院院刊,2000:24(1):49
[3]Kwang-Youl Ryu,Won-Seek Kang,Young-Ho kim.Antioxidative Effects of the Rhizome of Rhodiola Sachalinensis.J.Pharm.Soc.korea,1998;42(3):312-318
[4]庞鹤,朱陵群,张文生,王硕仁,牛福玲,崔巍.红景天苷对缺氧缺糖损伤的神经细胞线粒体膜电位和凋亡的影响.中医药学刊,2005;23(12):2144
[5]张桂霞,李立,张杰.红景天缓解体力疲劳作用的实验研究.医药论坛杂志,2005:26(23):48-49
[6]徐峰,陈星,韩璐璐.对红景天抗疲劳作用机理的探讨.食品科学,2004;25(10):366-366
[7]孙非,王秀清,李静波.高山红景天多糖抗柯萨奇B5病毒作用的试验研究.中草药,1993;24(10):532
[8]肖辉,张月明.红景天提取物的抗肿瘤实验.中国临床康复,2006;70:142
[9]盛辉,苑春莉,李洋.红景天对体外培养肺腺癌细胞增殖的抑制作用.中国实验诊断学,2005;9(6):90-91
[10]马玉华.红景天治疗心血管疾病研究进展.中国分子心脏病学杂志,2002:2(4):52-56
[11]张维烨,李艳君,钟震亚,王守安,陈立强.红景天苷对神经干细胞作用的研究.黑龙江医药科学,2005;28(6):3-4
[12]张文生,朱陵群,牛福玲.红景天苷对缺氧/缺糖损伤神经细胞的保护作用.中国中药杂志,2004;25(8):459-461
[13]金沈锐,秦旭华.红景天提取物对初老大鼠海马中神经生长因子和脑源性.神经营养因子含量影响.中国中药杂志,2004;29(5):480-481
[14]Zhang L,Yu H,Sun Y,Lin X,Chen B,Tan C,Cao G,Wang Z.Protective effects of salidroside on hydrogen peroxide-induced apoptosis in SH-SY5Y human neuroblastoma cells.Eur J Pharmacol.2007;564(1-3):18-25
[15]张汝学,王玉芝.红景天多糖对小鼠体内脾淋巴细胞转化反应及NK细胞活性的影响.中国药理学通报,1993;9(4):278
[16]胡晓波,杨礼腾,穆茂,聂莉,方洵,李群,程德云.红景天对大鼠肺纤维化治疗作用初探.四川医学,2006;27(7)673-674
[17]张奕,刘永冈.红景天苷抗肝纤维化的实验研究.中国药房,2006;17(11):813-815
[18]姜义,肖雪媛,关桂梅.红景天素对X射线照射小鼠的预防作用.中华放射医学与防护杂志,1995;15(3):214
[19]贾正平.谢景文,魏虎来.狭叶红景天水提物的辐射保护作用.兰州医学院学报,1997;23(3):17-19
[20]Kim SH,Hyun SH,Choung SY.Antioxidative effects of Cinnamomi cassiae and Rhodiola rosea extracts in liver of diabetic mice.Biofactors.2006;26(3):209-19
[2]Subramoniam,A.,Too many hepatoprotective drugs and a little hepatoprotection.Current Science,1995,69,898-899
[3]王亮,游冬青,陈杞.红景天药理研究进展.时珍国医国药,1999,10(4):296-296
[4]高学敏主编.中药学.第1版.北京:中国中医药出版社,2002:511
[5]高秋娜.红景天的药理与临床研究概述.河南中医,2001,22(3):76-77
[6]Min-Won Lee,Yeon-Ah Lee,Hyang-Mi Park,Sang-Hak Toh,Eun-Joo Lee,Hae-Dong Jang,Young-Ho Kim.Antioxidative phenolic compounds from the Roots of Rhodiola sachlinensis A.Bor.Medicinal Chemistry & Natural Products,2000,23(5):455-458
[7]Lei Chen,Hong-Yu Zhang.Cancer Preventive Mechanisms of the Green Tea Polyphenol(-)-Epigallocatechin-3-gallate.Molecules,2007,12:946-9
[8]Nan JX,Jiang YZ,Park EJ,Ko G,Kim YC,Sohn DH.Protecive effect of Rhodiola sachalinensis extract on carbon tetrachloride-induced liver injury in rats.Journal of Ethno-Pharmacology,2003,84:143-148
[9]Song EK,Kim JH,Kim JS,Cho H,Nan JX,Sohn DH,Ko GI,Oh H,Kim YC.Hepatoprotective phenolic constituents of Rhodiola sachalinensis on tacrine-induced cytotoxicity in Hep G2 cells.Phytother Res.,2003,17:563-565
[10]Williams,R.Classification,etiology,and considerations of outcome in acute liver failure.Semin.Liver Dis,1996;16:343-348
[11]Ostapowicz,G.,and W.M.Lee.Acute hepatic failure:a Western perspective.J Gastroenterol.Hepatol,2000;15:480-488
[12]Galanos,C.,Freudenberg,M.A.,Feutter,W..Galactosamine-induced sensitization to the lethal effects of endotoxin.Proc.Natl.Acad.Sci. U.S.A,1979;76:5939-5943
[13]WANG H,XU DX,Lu JW,Zhao L,Zhang C,Wei W.N-Acetyl cysteine attenuates lipopolysaccharide-Induced apoptotic liver damage in D-galactosamine-sensitized mice.Acta Pharmacol Sin,2007;28(11):1803-1809
[14]Tetsuko Fulkuda,Akira Mogami,Hideki Tannaka,Tsutomu Yoshikawa,Masao Hisadome,Hirotsugu Komatsu.Y-40138,amultiple cytokine production modulator,protects against D-galactosamine and lipopolysaccharide-induced hepatitis.Life Sciences,2006;79:822-827
[15]Anne M.Larson,MD,FACP,AGAF.Acetaminophen Hepatotoxicity.Clin Liver Dis,2007,11:525-548
[16]O Brien,J.T.,Eagger,S.,Levy,R..Effects of tetrahydroaminoacridine on liver function in patients with Alzheimer's disease.Age Ageing,1991,20:129-131
[17]Arvelo,M.B.,Cooper,J.T.,Longo,C.,Daniel,S.,Grey,S.T.,Mahiou,J.,Czismadia,E.,Abu-Jawdeh,G.,Ferran,C..A20 protects mice from D-galactosamine/lipopolysaccharide acute toxic lethal hepatitis.Hepatology,2002,35:535-43
[18]Huiping Wang,Yan Li.Protective effect of bcyclol on acute hepatic failure induced by lipopolysaccharide and D-galactosamine in mice.European Journal of Pharmacology,2006,534:194-201
[19]雷桂兰,吴中华,刘世旺,李树文.苦荆茶中多酚类物质的含量测定.长江大学学报,2005,1(2):37-39
[20]Sung-Hwa Kim1,Yeong Shik Kim,Sam Sik Kang,KiHwan Bae,Tran Manh Hung,and Sun-Mee Lee1.Anti-apoptotic and Hepatoprotective Effects of Gomisin A on Fulminant Hepatic Failure Induced by D-Galactosamine and Lipopolysaccharide in Mice.J Pharmacol Sci,2008,106:225-233
[21]Brouckaert P,Fiers W.Tumor necrosis factor and the systemic inflammatory response syndrome.Curr Top Microbiol Immunol,1996;216:167-187
[22]田静.水飞蓟素抗肝损药理及其复合物的研究进展.海峡药 学,2004,16(5):7-9
[23] Mary Lynn Bajt, Anwar Farhood. Mitochondarlal Bax Translocation Accelerats DNA Fragmentation and cell necosis in murine model of acetaminophen hepatototoxicity. American Society for Pharmacology and Experimental Therapeutics, 2007,10:1-25
[24] Jain A, Soni M, Deb L, Jain A, Rout SP, Gupta VB, Krishna KL. Antioxidant and Hepatoprotective activity of ethanolic and aqueous extracts of Momordica dioica Roxb. Leaves. Journal of Ethno-Pharmacology, 2008,115:61-66
[25] Knight TR, Kurtz A, Bajt ML, Hinson JA, Jaeschke H. Vascular and hepatocellular peroxynitrite formation during acetaminophen toxicity: role of mitochondrial oxidant stress. Toxicol Sci, 2001, 62:212-220
[26] Laura P. James, Brian Donahower, Angela S., Burke, Sandra McCullough, Jack A. Hinson. Induction of the nuclear factor HJP-1a in acetaminophen toxicity: Evidence for oxidative stress. Biochemical and Biophysical Research Communications, 2006,3:171-176
[27] Stachlewitz RF, Arteel GE, Raleigh JA, Connor HD, Mason RP, Thurman RG. Development and Characterization of a New Model of Tacrine-Induced Hepatotoxicity: Role of the Sympathetic Nervous System and Hypoxia-Reoxygenation. The Journal of Pharmacology and Experimental, Therapeutics, 1997, 282(3): 1591-1599
[1]陈纪军等.德钦红景天的化学成分.云南植物研究,1999:21(4):525
[2]田军等.红景天挥发油的化学成分研究.军事医学科学院院刊,2000:24(1):49
[3]Kwang-Youl Ryu,Won-Seek Kang,Young-Ho kim.Antioxidative Effects of the Rhizome of Rhodiola Sachalinensis.J.Pharm.Soc.korea,1998;42(3):312-318
[4]庞鹤,朱陵群,张文生,王硕仁,牛福玲,崔巍.红景天苷对缺氧缺糖损伤的神经细胞线粒体膜电位和凋亡的影响.中医药学刊,2005;23(12):2144
[5]张桂霞,李立,张杰.红景天缓解体力疲劳作用的实验研究.医药论坛杂志,2005:26(23):48-49
[6]徐峰,陈星,韩璐璐.对红景天抗疲劳作用机理的探讨.食品科学,2004;25(10):366-366
[7]孙非,王秀清,李静波.高山红景天多糖抗柯萨奇B5病毒作用的试验研究.中草药,1993;24(10):532
[8]肖辉,张月明.红景天提取物的抗肿瘤实验.中国临床康复,2006;70:142
[9]盛辉,苑春莉,李洋.红景天对体外培养肺腺癌细胞增殖的抑制作用.中国实验诊断学,2005;9(6):90-91
[10]马玉华.红景天治疗心血管疾病研究进展.中国分子心脏病学杂志,2002:2(4):52-56
[11]张维烨,李艳君,钟震亚,王守安,陈立强.红景天苷对神经干细胞作用的研究.黑龙江医药科学,2005;28(6):3-4
[12]张文生,朱陵群,牛福玲.红景天苷对缺氧/缺糖损伤神经细胞的保护作用.中国中药杂志,2004;25(8):459-461
[13]金沈锐,秦旭华.红景天提取物对初老大鼠海马中神经生长因子和脑源性.神经营养因子含量影响.中国中药杂志,2004;29(5):480-481
[14]Zhang L,Yu H,Sun Y,Lin X,Chen B,Tan C,Cao G,Wang Z.Protective effects of salidroside on hydrogen peroxide-induced apoptosis in SH-SY5Y human neuroblastoma cells.Eur J Pharmacol.2007;564(1-3):18-25
[15]张汝学,王玉芝.红景天多糖对小鼠体内脾淋巴细胞转化反应及NK细胞活性的影响.中国药理学通报,1993;9(4):278
[16]胡晓波,杨礼腾,穆茂,聂莉,方洵,李群,程德云.红景天对大鼠肺纤维化治疗作用初探.四川医学,2006;27(7)673-674
[17]张奕,刘永冈.红景天苷抗肝纤维化的实验研究.中国药房,2006;17(11):813-815
[18]姜义,肖雪媛,关桂梅.红景天素对X射线照射小鼠的预防作用.中华放射医学与防护杂志,1995;15(3):214
[19]贾正平.谢景文,魏虎来.狭叶红景天水提物的辐射保护作用.兰州医学院学报,1997;23(3):17-19
[20]Kim SH,Hyun SH,Choung SY.Antioxidative effects of Cinnamomi cassiae and Rhodiola rosea extracts in liver of diabetic mice.Biofactors.2006;26(3):209-19