摘要
目的:观察“肝心宁”对实验大鼠肝纤维化的防治作用,研究“肝心宁”对实验大鼠肝纤维化的防治作用的机理,为“肝心宁”在临床的广泛应用提供理论依据。
方法:57只sD大鼠随机分为空白组、模型组、阳性对照组、中药低剂量组、中药高剂量组5组。采用改良复合因素法复制大鼠肝纤维化模型。空白组与模型组给予生理盐水10ml/kg灌胃,阳性对照组给予秋水仙碱0.125mg/kg治疗,中药高、低剂量组分别给予肝心宁26g/kg、6.5g/kg治疗。6周后,HE染色后比较各组大鼠肝脏病理组织改变;采用全自动生化分析仪,对各组大鼠血清ALT、AST、TBA进行检测;采用放免分析法检测大鼠血清HA、PⅢNP、LN的含量;用免疫组织化学法研究各组大鼠肝脏组织中caspase-3、bcl-2表达的部位及强度;并测定肝组织中PDGF-BB、bFGF的表达情况。
结果:在肝脏病理积分上各治疗组与模型组比较均有显著性差异(P＜0.05或P＜0.01),且在炎症积分上中药低剂量组效果最为明显,与阳性对照组比较也有显著性差异(P＜0.05);在血清酶学指标上,与空白组比较,其余各组大鼠血清中ALT、AST的活性与TBA的含量均明显升高,有显著性差异(P＜0.01或P＜0.05)。与模型组比较,各治疗组血清中ALT、AST的活性与TBA的含量均明显减少有显著性差异(P＜0.01或P＜0.05),各治疗组间无显著性差异(P＞0.05);在血清ECM指标上,与空白组比较,其余各组大鼠血清中LN、HA、PⅢNP的含量均明显升高,有显著性差异(P＞0.01或P＜0.05),与模型组比较,各治疗组血清中LN、HA、PⅢNP的含量均明显减少,有显著性差异(P＜0.01或P＜0.05)。治疗药物之间,除血清LN中药低剂量组显著低于阳性对照组(P＜0.05)外,其余各治疗组之间比较无显著性差异(P＞0.05);在抑制bFGF表达上,与模型组比较,各治疗组肝组织中bFGF均明显减少,有显著性差异(P＜0.01),各治疗组间无显著性差异(P＞0.05);在抑制PDGF-BB表达上,各治疗组与模型组比较均有显著性差异(P＜0.01),各治疗组间无显著性差异(P＞0.05);在调节caspase-3方面,与模型组比较,各治疗组肝组织中caspase-3均明显减少,有显著性差异(P＜0.01或P＜0.05)。,治疗药物之间,中药低剂量组显著低于阳性对照组(P＜0.05),肝心宁低剂量组、肝心宁高剂量之间比较无显著性差异(P＞0.05);在调节bcl-2方面,与空白组比较,其余各组各组大鼠肝组织中bcl-2均明显升高,有显著性差异(P＜0.01),与模型组比较,各治疗组肝组织中bcl-2未见明显减少(P＞0.05)。
结论:“肝心宁”能有效预防实验大鼠肝纤维化形成,改善肝纤维化大鼠肝组织损伤程度,可以有效降低肝纤维化模型大鼠血清ALT、AST、TBA等酶学指标和LN、HA、PⅢNP等肝纤维化指标的含量。其机制可能与“肝心宁”能够抑制PDGF-BB、bFGF介导的HSC增殖有关;此外通过调整caspase-3,bcl-2减轻肝细胞凋亡也可能“肝心宁”防治肝纤维化机制之一。中药防治肝纤维化往往是通过多途径、多层次、多靶点来实现的,所以以上结论的可靠性还需要在以后的科研临床工作中进一步验证。
Objective:To observe the preventive and therapeutic effect of "Ganxinning" capsule on experimental HF rats and the possible therapeutic mechanism,to provide the theoretic evidence for the clinic apply of"Ganxinning".
Methods:57 SD rats were divided into 5 groups randomly,the normal group,the model group,the positive control group,the high dose group,and the low dose group.HF rat model was duplicated with Improved Composite Factor Method.Rats of the normal group and the model group were administered normal sodium with dose 10ml/kg,positive control group were administered Colchicine with the dose 0.125mg/kg.The high dose group and the low dose group were administered "Ganxinning" with the dose 26g/kg、6.5g/kg respectively.The liver pathology change of each group by HE coloration were compared and determination of liver tissue PDGF-BB expression 6 weeks later.The ALT,AST,and TBA in blood serum of experimental rats were detected by automatic biochemical analyzer.The HA,PⅢNP and LN were detdected by radio-immunoassay.The expression of caspase-3,bcl-2,PDGF-BB and bFGF were detected by immunohistochemical analyzer,
Results:Compared with the model group,each treatment group has significant difference in the liver pathology integral(P＜0.05 or P＜0.01),and the low dose group is the most significant difference in the inflammation integral.Compared with the Colchicin group,each treatment group has significant difference too(P＜0.05).Compared with the control group,the ALT,AST and TBA of each group increased significantly(P＜0.05 or P＜0.01).And compared with the model group,these indexes of each treatment group decreased significantly(P＜0.01 or P＜0.05).these indexes between each treatment group showed no significant difference(P＞0.05).Compared with the control group,the LN,HA and PⅢNP of each group increased significantly(P＜0.05 or P＜0.01).And compared with the model group, these indexes of each treatment group decreased significantly(P＜0.01 or P＜0.05).The LN of lower dose TCM group was lower significantly than the Clochicin group(P＜0.05).these indexes between the other treatment groups showed no significant difference.Compared with the model group,the expression of bFGF of each treatment group decreased significant(P＜0.01).the expression of bFGF between each treatment group showed no significant difference(P＞0.05).Compared with the model group,the expression of PDGF-BB of each treatment group decreased significant(P＜0.01).the expression of PDGF-BB between each treatment group showed no significant difference(P＞0.05).Compared with the model group,the caspase-3 in olive of each treatment group decreased significant(P＜0.01 or P＜0.05). The caspase-3 of lower dose TCM group was lower significantly than the Clochicin group(P＜0.05).The caspase-3 of the higher Ganxinning dose and lower Ganxinning dose showed no significant difference(P＞0.05).Compared with the control group,the bcl-2 of each treatment group increased significant(P＜0.01).Compared with the model group,thebcl-2 between each treatment group showed no significant difference(P＞0.05).
Conclusion:"Ganxinning"capsule can prevent HF of the experimental rats effectively and decrease the enzymology index of ALT,AST,TBA and the HF index of LN,HA,PⅢNP.The possible mechanism is that Ganxinning can depress the HSC multiplication which were transfected by PDGF and bFGF. The other possible mechanism is that it can adjust the caspase-3、bcl-2 to prevent the apoptosis of liver cell.The TCM was used on HF mostly through multi-approach,multi-hiberarchy and multi-target.All of these experiment data need to be verified by clinical study.
引文
[1]Montfort I,Preze-TamayoR.Collagenase in experimental carbon tetrachloride cirrhosis of the liver.Am J Pathol,1997,92:411.
[2]刘平.高云华.肝纤维化动物模型的建立.世界华人消化杂志,2002,10(6):693.
[3]韩德五,马学惠,赵元昌.肝硬化动物模型的研究.山西医药杂志,1979,(4):1.
[4]李生财,李形.肝纤维化动物模型的造模原理及应用.中医药学刊,2006,24(12):2267-2268
[5]李少林.核医学[M].北京:人民卫生出版社,2002:33.
[6]中化医学会肝病学分会肝纤维化学组.肝纤维化诊断及疗效评估共识 中化肝脏病杂志,2002,10:327-328.
[7]中华医学会肝病学分会脂肪肝和酒精性肝病学组.酒精性肝病诊疗指南.中华肝脏病杂志,2006,3:164-165.
[8]Montfort I,Preze-Tamayo R.Collagenase in experimental carbon tetrachloride cirrhosis of the liver.Am J Pathol,1997,92:411.
[9]Jiang Z,You D Y,Chen X Cetal.Monitoring of serum markers of fibrosis during CC14-induced liver damage-effects of antifibrotic agents.J Hepatol,1992;16(3):282-284.
[10]3Jonker AM,Dijkhuis F,Hardonk MJ,et al.Immunohistochemical study of hepatic fibrosis induced in rats by multiple galactosamine injections[J].Hepatology,1994,19:775-781.
[11]牛挺献,史智勇.肝纤维化动物模型的研究.实验动物科学与管理,1997,14(3):27.
[12]刘平,高云华.肝纤维化动物模型的建立.世界华人消化杂 志,2002;10(6):693-5.
[13]汪国运,蔡卫民,等.肝纤维化组织学量化诊断的实验研究.中华肝脏病杂志,1998,6(4):56-58.
[14]Sugiyama M,Ichhida T,sato T,et al.Expression of activinA is increased in cirrhotic and fibrotic rat livers.Gastroenterology,1998,114:550.
[15]王宝恩,王志富,殷蔚荑,等.实验性免疫性肝纤维化模型的研究.中华医学杂志,1989.69(9):503.
[16]李培建,梁保忠,王爱民,等.四氯化碳、人血白蛋白所致大鼠肝纤维化动物模型的实验研究.实验动物科学与管理,2000,17(2):32.
[17]Tams EG.Morphoiogical and functional changes in the livers of rats after ligation and excision of the common bile duct.Am J pathol,1957,33:13.
[18]Dunn MA,saber MA,Warren KS,et al.Liver Collagen Synthesis in murine schistosomiasis.J Clin invest,1977,59:666.
[19]韩德五,马学惠,赵元昌.肝硬化动物模型的研究.山西医药杂志,1979,(4):1.
[20]孙敬方主编.动物实验方法学.北京:人民卫生出版社,2001.474-475.
[21]陆伦根.曾民德.范建高,等.实验性肝纤维化组织中细胞间粘附分子表达的研究.中华內科杂志,1999,68(1):37.
[22]范建高,曾民德,李继强.等.肝内脂肪和脂质过氧化与肝纤维化关系的实验研究,中华内科杂志,1997,36(12):808.
[23]Bataller R,Brenner D A.Liver fibrosis[J].J Clin Invest,2005,115:209.
[24]Liaw YETai DI,Chu CM,et al.The development of cirrhosis in patients with chronic type B hepatitis:a prospective study.Hepatology,1988,493-496.
[25]Hsu YS,Chien RN,Yeh CT,et al.Long-term outcome after pontaneous HBeAg seroconversion in patients with chronic epatitis B.Hepatology,2002,35:1522-1527.
[26]Wu GC,Zhou W只Zhao YR,et al.Study on the natural history of chronic hepatitis B.Zhonghua Ganzangbing Zazhi,2002,10:46-48.
[27]Becket U.et al.Hepatology 2002,35:868-875.
[28]Michael PM,Christian PS.Autoimmune hepfifis:dinical challenges.Gastroenterology,2001,120:1502.
[29]Yabushita K,Yabushita K.Aberant expression of cktokeratin7 as a histological marker of progression in primary biliary cirrhosis.Liver,2001,121(1):50.
[30]Ahern M,Hall P,Halliday J.Hepatic stellate cells nomenclature(letter)[J].Hepatology,1996,23(1):193.
[31]Geerts A.History heterogeneity developmental biology and functions of quiescent hepatic stellate cells.Semin Liver Dis.2001,21:311-335.
[32]Jean S,Campbell Steven D,Hughs Debra G,et at.Pletelet derived growth factor C induces liver fibrosis,steatosis,and hepatocellular carcinoma[J].Proe Natl Acad Sci.2005,102(9):3389-3394.
[33]Gressner AM.The up and down of hepatic stellate cells in tissue injury:apoptoosis restores cellular moneostasis[J].Gastroenterplogy,2002,120:1285.
[34]Senon H,Structure and function of hepatic stellate cells[J].Med Electron Microsc,2004,37(1);3-15.
[35]Burt AD.Cellularand molecularaspects Of thepatic fibrosis[J].J Patbol,1993,170:105-114.
[36]Schebesch C,Kodel ja V,Muller C,et al.Alternatively activated macrophages actively inhibit proliferation of peripheral bloed lymphoeytes and CD4+ T cells in vitro[J].Immunology,1997,92:478-86.
[37]Muriel P,Escobar Y,Kupfer cells are responsible for liver cirrhosis induced by carbon tetrachloride[J].J Appl Toxicol,2003,23:103-108.
[38]Hironaka K,Sakaida I,Matsmura et al.Enhanced interstitial collagenase production of Kupfer cell by gadolinium chloride prevents pig serunirduced rat liver fibrosis[J].Biochem Biophys Res Commun,2000,267:290-295.
[39]Xidakis C,Ljumovic D,Manousou P,et al.Production of pro and an tifibrotic agents by rat Kupfer cells;the efect of octreotide[J].Dig Dis Sci,2005,50:935-941.
[40]Fiseher R,Cariers A,Reinehr R,et al.Caspase 9-dependent killing of hepatic stellate cells by activated kupfer cells[J].Gastroenterology,2002,123:845-861.
[41]Xaus J,Comalada M,Valledor AF,et al.LPS induces apoptosis in macrophages mostly through the autocrine prod uction of TNF-alpha[J].Blood,2000,95:3823-3831.
[42]Sokol RJ.Liver cell injury and fibrosis[J].Pediat Gastroenterol Nutr,2002,35:7-10.
[43]Keynolds WF,Patel K,Pianko S.A genotypic association implicates myelope roxidase in the progression of hepatic fibrosis in chronic hepatitis Cvirus infection[J].Genes lmmun,2002,3:345-349.
[44]Van Eyken P,Sciot R,Desmet VJ,et al.Expression of the noevl extra- cellular matrix component tenascin in normal and diseased human liver.An immunohisto-chemical stydy[J].J Hepatol,1990,11(1):42-52.
[45]Kim T,Mars W,Stolz D,et al.Expression and activation Of pro-MMP-2 and pro-MMP-9 during rat liver regeneration[J],Hepatology,2000,31(1):75-82.
[46]Intmaka S,Uano T,Torimura T,et al.The signif icance of colocal ization of plasminogen activator inhibitor-1and vltronect in in hepatic fibrosis[J].Scand J Gastroenterol,1997,32(10):1052-1060.
[47]Kanzler S,Lohse AW,Keil A,et al.TGF-β in liver fibrosis:an in dueible transgenic mouse model to study liver fibrogmesis[J].Am J Physiol,1999,276(4):1059-1068.
[48]王爱民,王宝恩,杨跃伟,等.表皮生长因子对肝星状细胞基质分解素-1及其抑制因子基因表达调节的影响[J].中国病理生理杂志,2002,18:197-198.
[49]Saile B,Nlatthes N,Knittel T,et al.Transforming growth factor beta and tumor necrosis factor alpha inhibit both apoptosis and proliferation of activated rat hepatic stellate cells[J].Hepatology,1999,30:196-202.
[50]Pinzani M,Marra F,Carloni V,et al.Signal transduction in hepatic stellate cells[J].Liver,1998,18:2-3.
[51]刘小菁.吴红斌,刘芳,等.肿瘤坏死因子-a对肝星状细胞表达结缔组织生长因子的影响[J].中华肝脏病杂志,2001,9:15-17.
[52]陈治新,张莉娟,陈运新,等.胰岛素样生长因子-1受体在实验性肝纤维化中的表达及IL-10的干预作用[J].福建医科大学学报,2002,36:134-136.
[53]黄新.激活素和纤维化[J].华人消化杂志,1999,19:158-160.
[54]Williams EJ,gaca MD,Brigstock DR,et al.Increased expression of connective tissue growth factor in fibrotic human liver and in activated hepatic stellate cells[J].J Hepatol,2000,32:754-761.
[55]Shao R,Rockey DC.Eflects of endothelins on hepatic stellate cell synthesis of endothelin-1 during hepatic wound healing[J].J Cell Physiol,2002,191:342-350.
[56]Diehl AM.Nonalcoholic steatosis and stetohepatitis Ⅳ.Nonalcoholic:fatty liver disease abnormalities in macro phage function and cytokines[J].Am J Physiol gastrointest Liver Physiol,2002,282:61-65.
[57]Ikejima K,Takei Y,Honda H,et al.Leptin receptor mediated signaling regulates hepatic profibrogenic and remodeling of extracellular matrix in the rat[J],gastroenterology,2002,122:1399-1410.
[58]扬永平.IFN-γ对肝星状细胞表达al(Ⅰ)mRNA信号传递途径的影响[J].中华肝脏病杂志,2000,8:123-124.
[59]Chinnaiyan AM,Tepper CU,Seldin MF,et al.FADD/MORT1 is a common mediator of CD95and tumor necrosis factor receptor induced apoptosis[J].J Biol Chem,1996,271:4961.
[60]Lonis H,Van Laethem JL,Quermrnom E,et al.Imerleukin-10 controis neutrophilic infiltraton,hepatoeyte proliferation,and liver fibrosis induced by carbon tetrachloride mice[J].Hepatology,1998,28(6):1607-1615.
[61]Thompaon KC,Maltby J,Fallowfield J,et al.Interleukin-10 expression and function in experimental murine liver inf lammation and fibrosis[J].Hepatology,1998,28(6):1597-1606.
[62]翁山耕,冷希圣,魏玉华.白细胞介素10抑制枯否细胞诱导的肝星状细胞激活的研究[J].中华医学杂志,2002,2:104-107.
[63]Rust C,Gores GJ.Apoptosis and liver disease[J].Am J Mbd,2000,106(7):567-574.
[64]彭黎明,王曾礼 细胞凋亡的基础与临床[M].北京:人民卫生出版社,2000,112-114,394-405.
[65]Canbay A,Friedman S,Gores GJ.Apoptosis:the nexus of liver injury and fibrosis[J],Hepatology,2004,39:273-278.
[66]Zhao YZ,Kim JY,Park EJ,et al.Tetrandrine induces apoptosis in hepatic stellate cells[J].Phytotlier Res,2004;18(2):306-309.
[67]YangJ,liuX,Bhalla K,et al.Prevention of apoptosis by Bcl-2:releaseof cytochrome c from mitochondria blocked[J].Science,1997,275:1129-1132.
[68]Bergsten E,IJutela M,li X.et al.PDGF-D is a specific,pmtease-activated ligand for the PDGF 13 receptor[J].Mat Cel Biol[J],2001,3:512-516.
[69]Pinzani M,Milani S,Herbst H,et al.Expression of platelet derived growth factor and its receptors in normal human liver and during active hepatic fibrogenesis[J].Am J Pathol,1996,148(3):785-800.
[70]Zhang BB,Cai WM,Weng HL,et al.Diagnostic value of platelet derived growth factor,transforming growth factorbetal,matrixmetalloproteinase-1,and tissue inhibitor of matrix metalloproteinase-1 in snlm and peripheral blood moaonuclear cells for hepatic fibrosis[J].World J Gastroenterol,2003,9(11):2490-6.
[71]Ikura Y,Morimoto H,Ogami M,et al.Expresion of platelet derived growth factor and its receptor in livers of patients with chronic liver disease[J].JGastroenterol,1997,32(4):496-501.
[72]Yoshi ji H,Noguchi R,Kuriyama S,et al.Imatinib mesylate attenuates liver fibrosis development in rats[J].Am J Physiol Gastrointest Liver Physiol,2005,288(5):907-13.
[73]Amakawa M,Endo Y.The motility of hepatic Ito cells can be acquired by their myofibroblastic transformation[J].Arch Histol Cytol,2002,55(2):169-78.
[74]Preaux AM,Mallat A,Rosenbaum J,et al.Pentoxiffy line inhibits growth and col lagen synthesis of cultured human hepatic myofibroblast like cells[J].Hepatology,1997,26(2):315-327.
[75]Caligiuri A,De Franco RM,Romaneili RG,et al.Antif brogenic effects of canrenone,an ant ial dosteronic drug,on hunan hepatic stellate cells[J].Gastroenterology,2003,124(2):504-520.
[76]Bolen JB,Brugge JS.Leukocyte protein tyrosine kinases,Potential targets for drug discovery[J].Annu Rev Immunol,1997,15:371-404.
[77]Mallat A,Gallois C,Tao J,et al.Platelet derived growth factor-BB and thrombin generate pos itire and negative signals for human hepatic stellate cell proliferation.Role of a prostaglandin/cyclic AMP pathway and crosstalk with endothelin receptor[J].J Biol Chem,1998,273(42):27300-27305.
[78]Garloni V,Pinzani M,Giusti S,et al.Tyrosine phosphory lation of focal adhesion kinase by PDGF is dependent on rats in human hepatic stellate cells[J],ttepatology,2000,31(1):131-140.
[79]Disario A,Bendia E,Svegliati Baron I,et al.Rearrangement of the cytoskeletal network induced by platelet derived growth factor in rat hepatic stellate cells,role of different intracellular signaling pathway[J].J Hepatol,2002,36(2):179-190.
[80]Benedetti A,Di Sario A,Casini A,et al.Inhibition of the NA(+) / H(+) exchanger reduces rat hepatic stelate cell activity and liver fibrosis,an in vitro and in vivo study[J].Gastroenterology,2001,120(2):545-56.
[81]刘海林,李宣海,王丹艺,等.肝纤维化时Ⅳ型胶原酶与金属蛋白酶组织抑制因子-1的表达[J].上海医学,2000,23(2):73-75.
[82]Kim R Bridle,Lin Li,Rosemary O Neill,et al.Coordinate activation of intracellular signlaing pathways by insulin like growth factor-1 and Platelet derived growth factor in rat hepatic stellate ceils[JJ.J Lab Clin Med,2006,147:234-241.
[83]Win KM,Charlotte F,Malat A,et al.Mitogenic effect of transforming growth factor-1 on human Ito cells in culture.evidence for mediation by endogenous platelet-derived growth factor[J].Hepatology,1998,18:137.
[84]Liu JY,Brass DM,Hoyle GM.et al.TNF receptor knockout mice are protected from the fibroproliferative effects of inhaled asbestos fibers[J].Am J Pathol,1998,153(6):1839-1847.
[85]Battegay EJ,Raines EW,Collbert T,et al.TNF stimulation of fibro blast proliferation dependence on platelet derived growth factor(PDGF)seeretion and alteration of PDGF receptor expression[J].Immu nol,1995,154:6040.
[86]Plomikov AN,Hubba SR,Schlessinger J,et al.Crystal structures of two FGF-FGFR complexes reveal the determinants ofligand-receptor specificity,Cell,2000,101:413-424.
[87]Gehrmam J,Jannes vievirs J,stewan L,et al.Differential expression of fibroblast growth factor-2 and receptor by glial cells in experimental autoimmune encephalomyelitis(EAE).J Glia 1996,16(2):93-100.
[88]陈佳,罗成群.碱性成纤维细胞生长因子(bFGF)与创伤愈合.中国烧伤亿疡杂志,2005,17:74-76.
[89]修日东,张学广.碱性成纤雏细胞生长因子治疗口腔黏膜溃疡的临床观察;河北医药,2003,25:637.
[90]严家春,陈文笔,马勇,等.慢性乙型肝炎肝组织中碱性成纤维细胞生长因子及其mRNA的表达.中华传染病学杂志,2004,22:314-317.
[91]付小兵,杨银辉,孙同柱,等.抑制或拮抗內源性bFGF活性对肠缺血再灌注所致肠、肝、肾功能的影响.中国危重病急救医学,2000,12:465-468.
[92]黄月红,王小众,陈运新,等.白细胞介素-10,血小板衍生生长因子-BB对肝星状细胞表达表皮生长因子和碱性成纤维细胞生长因子的影响.中华肝脏病杂志,2004,12:562-563.
[93]张愚,屈文东,黄华,等.bPGF在实验性肝硬化中的免疫组化研究.胃肠病学和肝病学杂志.2005.14:40-44.
[94]辛邵杰,邹正升.细胞外基质的代谢(J).世界华人消化杂志,2002,10(1):54-6.
[95]Monfort I.Collagenase in experimental carbon tetraehloride cirrhosis of the liver[J].Liver J Pathol,1978,92:411-415.
[96]王晋源,.雷教华.乙型肝炎辩证分型与检测指标关系的探讨[J].现代中医,1990,4(1):31.
[97]薛博瑜,顾学兰,肝纤维化的病机认识和辩证论治[J].南京中医药大学学报(自然科学版),2001,17 C):76.
[98]牛建昭,李健.等.酒精性肝纤维化“毒损肝络”病机假说的形成及临床意义[J].中西医结合且病杂志,1996,6(5):46.
[99]Hoyumpa AM,Schenker S.Is glucu ronidation truly preserved in patients with Liver disease[J],neoatology,1991,13:786-795.
[100]叶任高,刘厚钰.內科学[M].北京:人民卫生出版社,2000,10.460.
[101]扬建辉.林鹤和治脂肪肝的经验[J].江西中医药,2000,31(3):1.
[102]Tokar JL,Berg CL.Therapeutic option in nonalcoholic fatty liver disease[J].Gurr Treat options Gastroenterol,2002,5(6):425-436.
[103]Jenta K,Van Thiel DH.nonalcoholic fatty liver disease Pathogenesis and the role pf antioxidants[J].Nu Rev,2002,60(9):289-293.
[104]郭丽珠.中成药致肝炎的病例分析[J].药物不良反应杂志年,2004,4:231-232.
[105]黄志余,林宪和.等.中医中药治疗抗结核药致药物性肝炎的观察[J].中国防痨杂志,2003,25(6):400.
[106]赵翠英,刘学华.茵黄合剂治疗妊娠肝內胆汁淤积症的实验研究[J].江苏中医,2000,7:39.
[107]马克明.扶正祛邪法治疗乙型肝炎150例[J].新中医,1996,28(9):46-47.
[108]张俊富,崔丽安,范淑芳,等.乙肝2号治慢性乙型肝炎的临床研究[J].中医杂志、1993,34(11):674.
[109]Lok hSF,Heathcote EJ,Hoofnagle JH.Management of hepatitis B:2000 summary of a workshop.Gastroenterology,2001,120:1828-1853.
[110]中华医学会肝病学分会,中华医学公感染病学分会.慢性乙型肝炎防治指南,2005年12月10日.
[111]Chisari FV,Ferrari CR.Hepatitis B virus immunopathogenesis Pe[J].Immunol,1995,13:29.
[112]唐智敏,茹清静.等.论肝血瘀阻与肝纤维化的关系[J].中国中医基础医学杂志,1996,2(3):14-18.
[113]于家军,李戈,等.李寿山教授治疗慢性肝炎的经验[J].辽宁中医杂志,1999,26(11):483-484.
[114]刘宏元,刘作恩.慢性乙肝早期肝纤维化的治疗研究[J].天津中医,1997,14(3):101-102.
[115]张浚富,崔丽安.慢性肝炎及肝硬化辩证分型与血清肝纤维化指标的关系[J].中西医结合肝病杂志,1995,5(4):10.
[116]崔丽安,张俊富,等.从慢性肝病胃粘膜的改变探讨中医“肝病传脾”理论的意义[J].中西医结合肝病杂志,1997,7(4):201-203.
[117]史广宇,单书健.当代名医临证精华.肝炎肝硬化专辑[M].北京:北京中医古籍出版社,1998.325.
[118]范宗滂.中药补肾养阴方抗肝纤维化的实验研究[J].江苏医药,1989,5:24.
[119]雀清萍,张延超,清扫化瘀方对慢性肝病肝纤维化及相关指标的影响[J].江苏中医,2000,21(11):9.
[120]于世瀛,清热利湿方抗肝纤维化的形态学和免疫组织化学定量研究[J].北京中医药大学学报,1997(3):26.
[121]王宝恩,孙淼,活血化瘀复方中药对实验性肝纤维化的疗效观察[J].中成药,1990,21(4):33.
[122]卢良威.张平,等.活血渗湿方抗肝纤维化作用的实验研究[J].渐江中医学院学报发,1999,23(4):47-48.
[123]晏丹,陈建明,等.水蛭桃仁汤对肝纤维化大鼠HSC活化及TGF-β表达的影响.中西医结合肝病杂志,2004,14(1):36-38.
[124]刘平.中医中药治疗慢性肝病的几个问题.中西医结合肝病杂志,2000,增刊:1-2.
[125]陈忠义.俞伟.慢乙肝患者肝组织胶原含量与中医征型关系的研究[J].中国医药学报,2000,15(3):73.
[126]张俊富,崔丽安.等.乙肝养阴活血冲剂抗肝纤维化的临床观察.天津中医,1996,13(3):14-1.
[127]苏涟,张超,等.健睥活柔肝法治疗慢性乙型肝炎257例疗效观察.中西医结合肝病杂志,1998,8:52-51
[128]陈文慧,王臻楠,等.福乐肝6号对DMN肝纤维化大鼠治疗作用的实验研究.中国中医药科技,2001,8:15-1.
[129]吴嘉庚,疏肝健脾活血法治疗乙型肝炎肝纤维化的研究[J],中国中西医結合杂志,1994,14(12):744.
[130]李倩,冯卫生.连翘的化学成分研究进展[J].河南中医学院学报,2005,20(11):78.
[131]芮菁,尾崎幸,唐元泰.连翘提取物的抗炎镇痛作用[J].中草药,1999,30(1):43.
[132]张海燕.连翘化学成分及药理活性的研究进展[J].中药材,2000,23(10):657.
[133]朱淑云,杨建雄,李发荣.连翘叶提取物对小鼠氧化损伤的保护作用[J].中药药理与临床.2004,20(1):18.
[134]李长城,严仪昭,等,连翘、丹参、和川芎嗪抑制弹性蛋白酶活力的对比观察[J].中国中药杂志.1990,15(9):47.
[135]雷载权.中药学[M].上海:上海科学技术出版社,1995,145-146.
[136]熊玉兰 茵陈有效成份对四氯化碳损伤的原代培养大鼠肝细胞的作用[J].中国实验方剂学杂志,2002,(1):32.
[137]昊其恺.中西医结合治疗病毒性黄疸型肝炎135例[J].湖北中医杂志,2000,(8):15-16.
[138]曾小玲.七种菊科植物抗活性缾作用的研究对起氧奶离子自由基的清除作用.中国现代医学杂志,1999;9(2):44.
[139]王新华.泽泻研究进展[J].中草药,1999,30(7):557.
[140]余世锋.黄贤樟.童光东.不同剂量丹参注射液抗肝纤维化临床研究[J].中国中西医结合杂志,2004,24(1):17-20
[141]Wasser S,Ho JM,Ang HK,et al.Salvia miitiorrhiza reduces experimentally induced hepatic fibrosis in rats[J].J Hepatol,1998,29(5):760-771.
[142]Lee TY,Mai LM,Wang GJ,et al.Protective mechanism of salvia miltiorrhiza on carbon tetracblorede induced acute hepatotoxicity in rats[J].J Pharmacol Sci,2003,91(3):202-210.
[143]刘永刚,陈厚昌,蒋毅萍.等.丹参酮ⅡA对四氯化碳致大鼠纤维细胞实验研究[J].中药材,2002,25(1):31-33.
[144]陈岳祥.丹参单体IH764-3抗肝纤维化作用的实验研究[J].中华医学杂志,1998,78(8):636-637.
[145]胡义扬,王润平,张学军,等,体外四氯化碳损伤肝细胞对肝星状细胞活化的影响及丹酚酸A 的干预效果[J].中华肝脏病杂志,2000.8(5)-299.
[146]王晓玲.刘平.等,丹酚酸A对成纤维细胞活力、增殖及胶原合成的影响[J].中西医结合肝病杂志,2000,10(1):22.
[147]刘成海,刘平,胡义扬,等.丹酚酸B盐刺激肝星状细胞活化与胞内信号转导的作用[J].2002 年全国中西医结合肝病学术会议论文汇编[A],贵阳.2002:162.
[148]Ping Liu,Yi-Yang Hu,Cheng Liu,et al,Clinic observation of soleianolic aeid B in treatment of liver fibrosis in chronic hepatitis B[J].World J Gastmenterol,2002,8(4):679.
[149]张明霞,李效忠,赵磊.红花抗衰老作用的实验研究[J].中草药,2001.32(1):52-53.
[150]柏惠英.Labruyere J,秦月琴.红花对脑减压缺氧缺血幼鼠脑神经元的保护作用[J].中草药,1992,23(10):531-533.
[151]杨丽华,张敏,马春.红花的现代研究进展[J].中国老年学杂,2007,27(7):1420-1430.
[152]王筠默.中药药理学[M].上海:上海科技出版社,1983:63.
[153]方文贤.医用中药药理学[M].北京:人民卫生出版社,1998:328.
[154]黄沛力,曾绍晖.银杏叶、山楂叶对氧自由基的清除作用[J].中国中药杂志,1996,21(4):245.
[155]曾绍晖,颂晓虹,赵彪 山楂叶茶提取物抗氧自由基作用的研究[J].首都医科大学学报,1996,21(14):245.
[156]陈建明,强世平,赵冬,等.水蛭桃仁汤抗肝纤维化的研究[J].武汉冶金科技大学学报,1998,21(3):356-359.
[157]李天全.新一代高效特异抗凝药物-水蛭素[J].生物医学工程学杂志,1998,15(3):306-310.
[158]晏丹,陈建明.水蛭桃仁煎剂抗实验大鼠肝纤维化机理研究[J].江苏中医药,2005,26(8):45-48.
[159]范秦鹤,朱爱华,吕兰熏.女贞子不同炮制品免疫作用比较[J].陕西中医学院学报,1999,22(2):34-35.
[160]赵英,闻杰,孙忠人.女贞子对小鼠脑、肝过氧化脂质含量及肝SOD活性的影响[J].中医药学报,1990,18(6):47.
[161]闫秀英.枸杞子对四氯化碳致大鼠肝脂类过氧化作用的抑制[J].包头医学院学报,1998;14(1):79.
[162]张敏.枸杞子抗衰老保肝等实验研究综述[J].时珍国医国药,2000;11(4):373-375.
[163]张明华,陈虹.五味子甲素和五味子醇甲对四氧化碳所致肝脏损伤的保护作用[J].武警医学,2002,13(7):395-396.
[164]包天桐,徐桂芳,等.五味子仁七种成分的一些药理作用比较[J].药学学报,1979,14(1):1.
[165]高普军.北五味子粗多糖保肝作用的机理[J].白求恩医科大学学报:自然科学版,1996,(1):23-24.
[166]刘宗香,胡觉民,张仲一.五味子的药理实验[J].天津中医,1986,3:48.
[167]苟贤筠,张伟莉,贾政海.猪苓多糖治疗乙型肝炎的研究进展[J].中华临床医学研究杂志,2005,11(5):680-681.
[168]朱自平,张明发,等.肉桂的温中止痛药理研究[J].中国中药杂志,1993,18(9):553-557.
[169]张明发,沈雅琴.肉桂的药理作用及温里功效[J].陕西中医,1995,16(1):39-42.
[170]冯书晓,陈文,等.甘草酸类制剂在肝病治疗领域的研究进展[J].农垦医学,2005,27(1):58-59.
[171]白木公康.甘草酸抗乙肝病毒的作用机理[J].和汉医药学杂志(日),1995;12(1):24.
[172]Ynshikawa M,Matsui Y,Kawamoto H,et al.Efects of glycyrrhizin on immune mediated eytotoxicity[J].J Gas troenterol Hepatol,1997,12(3):243-248.
[173]郝飞.甘草酸国外研究进展[J].中国药房.2001,12(8):500-501.
[1]余世锋,黄贤樟,童光东.不同剂量丹参注射液抗肝纤维化临床研究[J].中国中西医结合杂志,2004,24(1):17-20.
[2]Wasser S,Ho JM,Ang HK,et al.Salvia miitiorrhiza reduces experimentally induced hepatic fibrosis in rats[J].J Hepatol,1998,29(5):760-771.
[3]Lee TY,Mai LM,Wang GJ,et al.Protective mechanism of salvia miltiorrhiza on carbon tetracblorede induced acute hepatotoxicity in rats[J].J Pharmacol Sci,2003,91(3):202-210.
[4]刘永刚,陈厚昌,蒋毅萍.等.丹参酮ⅡA对四氧化碳致大鼠纤维细胞的实验研究[J].中药材,2002,25(1):31-33.
[5]陈岳祥.丹参单体IH764-3抗肝纤维化作用的实验研究[J].中华医学杂志,1998,78(8):636-1637.
[6]胡义扬 王润平,张学军,等,体外四氯化碳损伤细胞对肝星状细胞活化的影响及酚酸的干预效果[J].中华肝脏杂志;2000,8(5)-299.
[7]王晓玲.刘平,刘成海,丹酚酸A对成纤维细胞活力、增殖及胶原合成的影响[J].中西医结合肝病杂志,2000,10(1):22.
[8]刘成海,刘平,胡义扬,等丹酚酸B盐刺激肝星状细胞活化与胞内信号转导的作用[J].2002年全国中西医结合肝病学术会议论文汇编[A],贵阳.2002:162.
[9]Ping Liu,Yi-Yang Hu,Cheng Liu,et al,Clinic observation of soleianolic aeid B in treatment of liver fibrosis in chronic hepatitis B[J].World J Gastmenterol,2002,8(4):679.
[10]石小枫 刘杞 刘林等.三七总苷抗实验性肝纤维化的研究[J].中药药理与临床,2004,20(1):12-13.
[11]张荣华,周子成,洪多伦,等.三七.人工虫草菌丝对实验性肝纤维化预防作用的研究[J].重庆医学,2000,29(6):483-486
[12]武凡,张树三,康格非.三七皂甙对肝纤维化大鼠分泌型磷脂酶A2和肿瘤坏死因子表达的影响[J].中华肝脏病杂志,2003,11(1):51-52
[13]方翠艳,杨金枝,富新伟,等.苦参素葡萄糖注射液治疗慢性乙型肝炎的疗效观察[J].实用肝脏病杂志,2004,7(4):238-239.
[14]王英,林小玲,张福华.苦参素对慢性乙型肝炎患者血清肝纤维化指标的影响[J].实用肝脏病杂志,2004,7(4):239-240.
[15]彭安邦,姚育红,谢黎明,等.苦参素治疗乙肝所致肝纤维化的临床观察[J].临床肝胆病杂志,2004,20(5):279-280.
[16]陈伟忠,张俊平.苦参碱对大鼠实验性肝纤维化的影响[J].第二军医大学学报,1996,17(5):424-426.
[17]卢清;张清波,张继明,等氧化苦参碱对大鼠肝星状细胞旁分泌活化途径的抑制作用[J].肝脏,2004,9(1):31-33.
[18]林明华,闰海明,李凯霞,等.氧化苦参碱对成纤维细胞增殖及Ⅰ型胶原合成的影响[I].黑龙江医药科学,2004,27(1):13-14
[19]甘乐文,王国俊,李玉莉.氧化苦参碱对大鼠肝纤维化的影响[J].第二军医大学学报,1999,20(7):447-448.
[20]Muriel P,Moreno MG.Effects of silymarin and vitamins E and C on liver damage induced by prolonged biliary obstruction in therat[J].Basic Clin Pharmacol Toxicol,2004,94(2):99-104.
[21]Jia JD,Bauer M,Cho JJ,et al.Antifibrotic effect of silymarin in rat secondary biliary fibrosis is mediated by downregulation of procol lagen alphal(Ⅰ) and TIg21[J].J Hepatol,2001,35(3):392-398.
[22]Schumann J,Prockl J,Kiemer AK,et al.Silibinin protects mice fron T cell dependent liver injury(small star,filled)[J].J Hepatol,2003,39(3):333-340.
[23]姬成伟,刘晋敏,李亮成,等.川芎嗪抑制大鼠肝纤维化的实验研究[J].山西医科大学学报,2003.31(4):293-295.
[24]王红,陈在忠,川芎嗪对大鼠肝纤维质过氧化的影响[J].中华肝脏病杂志,2000,8(2):98.
[25]展玉涛,李定国,等.大黄素抗纤维化的细胞学机制[J].临床肝胆病杂志,2006.22(1):13-44.
[26]展玉涛,魏红山,王志荣,等.大黄素抗肝纤维的实验研究[J].中华肝脏病学杂志,2001,9(4):235-239.
[27]陆江,李辰蕊,黄芪注射液对早期肝硬化患者生化及肝纤维化指标的响[J].山东中医杂志,2003,22(12):714-715.
[28]程鹏,刘素侠,孙晋浩,等.黄芪抗肝纤维化的作用与TGFβ的关系[J].临床军医杂志,2000,28(3):22-23.
[29]马红,王宝恩,黄芪抑制大鼠肝星状细胞增殖及胶原分泌的研究[J].中西医结合肝病杂志,2000(增刊):97-98.
[30]王要军,权启镇,孙自勤等.黄芪对实验性肝纤维化组织ICAM-1表达影响的免疫组化研究[J].中国临床药理学与治疗学,2000,5(1):49-51.
[31]韩涛,王宝恩,钱绍诚 黄芪对肝星状细胞胶原降解基因表达的影响[J].天津医科大学学报,2003,9(2):159-161.
[32]吴建良,刘成海 冬虫夏草对肝纤维化的作用[J].中西医结合肝病杂志,2001,11(6):382-384.
[32]颜吉丽,李华,范钰,等.虫草多糖对大鼠肝星状细胞NF-KB活性表达的影响[J].复旦学报(医学版),2003,30(1):27-29.
[33]刘玉侃,沈薇,张霞.虫草菌丝对实验性肝纤维化的防治作用及其机制研究[J].中国药与临床杂志,2004,23(3):139-143.
[34]Liu YK,Shen W.Inhibitive effect of cordyceps Sinensis on exprimental hepatic fibrosis and its possible mechanism[J].Wrld J Gastroenterol,2003,9(3):529-533.
[35]王要军,权启镇,孙自勤,等.复方汉防己治疗实验性肝纤维化的机理探讨[J].中国中西医结合脾胃杂志,200,8(30):151 152
[36]陈颖伟,李定国,等.汉防己甲素抗肝纤维化研究进展[J].中华消化杂志,2000,20(1):47-48.
[37]王伯祥,张赤志,聂广.肝胆病中西医诊疗学[M].北京:中国中医药出版社,2000,571-574.
[38]Park P H,Nan J x.Park E J,et al.Elect of tetrandrine on expefi mental hepatic fibrosis induced by bile duct ligation and scission in rats[J].Pharmacol Toxical,2000,87(6):261.
[39]王志荣,陈锡美,李定国,等,粉防己碱抑制肝纤维化大鼠肝组织c-fos和c-jun mRNA表达[J].上海医学,2003,26(5):332.
[40]Lee S H,Man J X,Sohn D H.Tetrandfine prevent tissue inhibitor of TIMP1mRNA expression in rat liver fibrosis[J].Pharmacol Toxicol,2001.89(6):214.
[41]袭柱婷,单长民,姜学连,等.三棱、莪术抗大鼠免疫性肝纤维化研究[J].中国中药杂志,2002,27(12):929-932.
[42]尹珊珊,王宝恩,王泰玲,等.复方861治疗慢性肝炎肝纤维化与早期肝硬化的临床研究[J].中华肝脏病杂志 2004,12(8):46-47.
[43]王宝恩,赵洪涛,王泰玲,等.复方861对肝纤维化疗效的病理学分析[J].中华肝脏病杂志,1997,5(2):77-78.
[44]马红,王宝恩,复方861对大鼠肝星状细胞增殖及胶原合成的影响[J].中国临床药理学与治疗学杂志,1998,3(3):172-174.
[45]周光德,李文淑,赵景民.等.复方鳖甲软肝片抗肝纤维化机制的临床病理研究[J].解放军医学杂志,2004,29:563-564.
[46]吴秀美,曾常春.鳖甲软肝片治疗慢性乙型肝炎肝纤维患者的临床观察[J].中药新药与临床药理,2003,14(3):203-205.
[47]Zeng MD,Lu LG,Mao YM,et al.Prediction of significant fibrosis in HBeAg-positive patients with chronic hepatitis B by a noninvasive model[J].World J Gastroenterol,2005,11:2892-2899.
[48]刘平,吴定中,刘成海等.扶正化瘀中药复方促进CCL4大鼠肝纤维化逆转的配伍机理研究[J].上海中医药大学学报.2002,16(1):37-41.
[49]宣红萍,孙保木,陶艳艳,等.肝纤维化过程中整合素动态变化与扶正化瘀方干预作用[J].肝脏,2004,9(3):163-166.
[50]姜春萌,刘成,刘成海,等.扶正化瘀方对大鼠CCL44损伤肝Kupffer细胞功能的影响[J].中西医结合肝病杂志,2000,10(5):26-28.
[51]季光,曹承楼,刘平,等.扶正化瘀方影响肝硬化大鼠肝细胞增殖的病理及免疫组化研究[J].中国中医基础医学杂志,2001,7(2):29-32.
[52]徐光福,陆雉,刘成海.扶正化瘀方对DMN诱导肝纤维化大鼠肝脏MMP-2活性的影响[J].肝脏,2002,7(增刊):85.
[53]崔红燕,姜哲浩,刘成,等.扶正化瘀方对肝纤维化大鼠间质性基质金属蛋白酶活性的影响[J].上海中医药大学学报,2003,17(3):35-38.
[54]刘成,姜春萌,刘成海,等.扶正化瘀方对肝星状细胞分泌血管内皮生长因子的干预[J].中华肝脏病杂志,2001,9(5):282-284.
[55]姜春萌,刘成海,刘成.扶正化瘀胶囊对肝星状细胞激活的干预[J].中国中西医结合消化杂志,2003,11(5):280-283.
[56]胡义扬,刘平,等.大黄遮虫丸抗慢性肝损伤及肝纤维化作用的实验研究[J].中西医结合肝病杂志,1995,5:28-30.
[57]林建辉,林应时,大黄遮虫丸对慢性肝炎肝纤维化指标的影响[J].中西医结合肝病杂志,2001,9(11):98-99.
[58]李岚生,徐曼,刘志龙,等.大黄遮虫丸对大鼠肝星状细胞表达TGFβ的影响.中国中西医结合杂志,2003,23(10):63-766.
[59]Shimizu I.Sho-saiko-to:Japanese herbal medicine for protection against hepatic fibrosis and carcinoma.J Gastroenterol Hepatol,2000,15 Suppl:D84-90.
[60]陈廷玉,王景涛,张波,小柴胡汤抗肝纤维化的形态学和血清学实验研究[J].黑龙江医药科学,2003,26(4):48-149.
[61]曾凡波,晏菊姣,万波等.鳖甲煎丸药理学研究[J].中成药,2002,24(7):529-532
[62]杨柳明,徐克成,赵延龙,等.强肝胶嚢治疗慢性乙型肝炎肝纤维化的临床病理研究[J].胃肠病学和肝病学杂志,2001,10:247-249
[63]王华,赵正龙,危北海,等.强肝胶囊联合拉米夫定治疗慢性乙型肝炎肝纤维化的临床病理观察[J].中国中西医结合杂志,2006,26(11):978-980.
[64]陈廷玉,卢春风,清云等.茵陈蒿汤对实验性肝纤维化大鼠肝细胞的保护作用及超微结构观察[J],黑龙江医药科学,2004,27(1):11-12.
[65]章以法.复方桃仁软胶囊治疗慢性乙型肝炎肝纤维化患者的临床观察[J].中国中西医结合杂志,2002,22(9):660-662.
[66]马晓峰,刘恩顺,高岚,大黄甘遂汤对小鼠实验性肝纤维化的防治作和研究[J].中医药学刊,2004,22(1):170-171.
[67]许佳平.双甲五灵胶囊治疗肝纤维化的Ⅰ晦床研究[J].中国中西医结合杂志,2002,10(3):159-161.
[68]陈跃,申碧荷,翁卫东,等.复方中药抗纤Ⅰ号抗肝纤维化的试验与临床研究[J].现代医学,2002,30(5):293-296.
[69]潘治平.健肝方治疗慢性乙型肝炎肝纤维化60例临床观察[J].湖南中医药导报,2002,8(6)}321-322.
[70]张国梁,软肝饮治疗慢性肝纤维48例疗效观察[J].安徽中医学院学报,1995,14(1):24.
[71]朱国静,吴建成,抗纤灵治疗慢性乙型肝炎临床观察[J].中华传染病杂志,1995,13(3):181.
[72]吴限,李岩,姜殿德,等.肝纤康合剂治疗肝纤维化的临床研究[J].哈尔滨商业大学学报,2002,18(1):105-106.