促肝细胞DNA合成刺激因了对大鼠肝纤维化的影响

目的：观察促肝细胞ＤＮＡ在刺激因子（ＨＤＳＳＦ）抗肝纤维化作用，探讨其可能的作用柚是。方法：采用复合病因法构建大鼠肝纤维化模型，以ＨＤＳＳＦ作为治疗药物，实验第１０周末取各组大鼠肝组织；（１）病理检查；（２）测肝纤维内羟脯氨酸含量；（３）Ｎｏｒｔｈｅｒｎ印迹法检测Ⅰ、Ⅲ型前胶原及胶原酶ｍＲＮＡ水平。结果：ＨＤＳＳＦ可明显改善肝组织的变性、坏死及纤维化程度、可使肝组织Ｈｙｐ含量明显下降，并可在减少Ⅰ     

肝纤维化组织学量化诊断的实验研究

目的探讨肝纤维化组织学量化诊断方法。方法用二甲基亚硝氨（ＤＭＮ）诱导ＳＤ大鼠肝纤维化模型，常规肝组织切片，Ｍａｓｏｎ三色和苦味酸天狼红染色，苦味酸固绿－天狼红染色比色测定，图像定量处理和半定量计分分析及羟脯氨酸（Ｈｙｐ）含量测定。结果切片胶原比色测定、图像定量处理和半定量计分分析均与肝纤维化程度变化相一致，与肝组织Ｈｙｐ含量具有良好的相关性，ｒ分别为０９４、０８９和０８８，Ｐ均＜００１。结论切片胶原测定、图像定量处理和半定量计分可作为确定肝纤维化程度的量化指标     

二甲基亚硝胺致大鼠脂质过氧化变化与药物干预作用

目的探讨二甲基亚硝胺（ＤＭＮ）诱导大鼠肝纤维化的脂质过氧化病理机制,以及扶正化瘀方与干扰素Ｖ的干预作用。方法ＤＭＮ复制大鼠肝纤维化模型,分别以扶正化瘀方灌胃及（干扰素肌注治疗。观察项目包括肝脏病理、肝功能、肝组织羟脯氨酸（Ｈｙｐ）、丙二醛（ＭＤＡ）含量与超氧化物歧化酶（ＳＯＤ）活性。并进行Ｈｙｐ、ＭＤＡ与ＳＯＤ之间的相关分析。结果模型大鼠肝组织胶原明显增生,纤维间隔形成; ＡＬＴ水平上升;肝组织Ｈｙｐ与ＭＤＡ含量显著增加, ＳＯＤ活性显著降低,且ＳＯＤ与 ＭＤＡ、 Ｈｙｐ呈负相关（ｒ＝０．７０,－０．７３,Ｐ＜ ０．０１）,ＭＤＡ与 Ｈｙｐ呈正相关（Ｔ＝０．８８, Ｐ＜ ０．０１）。扶正化瘀方与干扰素γ均能明显减轻肝组织胶原沉积、降低ＡＬＴ水平,提高肝组织ＳＯＤ活性、降低肝组织Ｈｙｐ与ＭＤＡ含量。结论脂质过氧化是ＤＭＮ损伤大鼠并诱发肝纤维 化的主要机制之一,抗脂质过氧化是扶正化瘀方与干扰素γ抗肝纤维化的重要作用机理。     

一种肝组织切片胶原定量分析法的实验性研究

目的　探讨肝纤维化组织切片上胶原测定方法。方法　用二甲基亚硝氨（ＤＭＮ） 诱导ＳＤ大鼠肝纤维化模型，常规肝组织切片，ＨＥ、Ｍａｓｓｏｎ 三色和苦味酸天狼红染色，苦味酸固绿天狼红染色比色测定及羟脯氨酸（Ｈｙｐ）含量测定。结果　对照组和肝纤维化明显组每张切片胶原含量分别为０．２３μｇ和１．０μｇ，非胶原蛋白含量分别为０．０６１ｍｇ 和０ ．０７１ｍｇ；切片胶原测定分别为３７ ．２３μｇ／ｍｇ·ｐｒｏ 和１２３．７μｇ／ｍｇ·ｐｒｏ；切片胶原与肝纤维化程度变化相一致，与肝组织Ｈｙｐ 含量具有良好的相关性，ｒ 均为０．９４ ，Ｐ＜０ ．０１ 。结论　切片胶原测定是一种简便、准确、可靠的肝纤维化程度的量化诊断方法。     

肝星状细胞与肝纤维化

1　肝纤维化的基本概念(1)肝纤维化是指各种原因所致的肝脏内纤维结缔组织增生,其特征为大量细胞外基质在Ｄｉｓｓｅ间隙沉积。(2)肝纤维化时不仅细胞外基质含量增加,而且细胞外基质的组成成分也发生了改变,正常肝脏Ｄｉｓｓｅ间隙细胞外基质是构成基底膜的成分,而肝纤维化时细胞外基质为形成纤维的胶原和纤维连接蛋白〔1〕。(3)在各种原因引起的肝纤维化中,肝脏细胞外基质含量均增加,但原因不同的肝纤维化早期,其肝脏增加的细胞外基质分布是不同的,如病毒性肝炎引起的肝纤维化主要分布在门脉区,而酒精引起的肝纤维化主要分布在中央静脉周围…     

白屈菜红碱对肝纤维化大鼠肝脏病理学和肝脏羟脯氨酸含量的影响

目的观察白屈菜红碱对四氯化碳诱导的肝纤维化大鼠肝脏病理学和肝脏羟脯氨酸（Hpy）含量的变化。方法采用四氯化碳联合营养控制和饮用10％酒精复合法制备SD大鼠肝纤维化模型,在实验第4周末,肝纤维化模型建立（2期）成功,然后应用不同剂量的白屈菜红碱和INF-γ处理。在给药8周后,处死所有大鼠,并取肝脏组织,采用HE、VG及Massion染色观察肝脏组织病理学形态的变化,同时采用改进的氯胺-T测定法检测肝脏Hyp含量。结果各剂量白屈菜红碱组肝纤维化半定量计分和肝脏的Hpy含量明显低于病理模型组（P〈0．01）;大中剂量白屈菜红碱组与小剂量组比,大鼠肝脏组织纤维化半定量计分及肝组织Hyp含量有明显下降,差异显著（P〈0．01）。结论白屈菜红碱可以改善肝纤维化大鼠肝组织纤维化程度,降低肝组织Hyp含量,具有抗化学性肝纤维化作用。     

珠子肝泰胶囊对大鼠肝纤维化的作用

目的：观察珠子泰胶囊（ＺＧＴ）的抗肝纤维化作用。方法：用四氯化碳（ＣＣｌ４）诱导大鼠肝纤维化模型,用ＺＧＴ治疗,观察其对大鼠肝功能、超氧化物歧化酶（ＳＯＤ）、丙二醛（ＭＤＡ）、透明质酸（ＨＡ）、Ⅲ型前胶原（ＰＣⅢ）、层粘蛋白（ＬＮ）及肝脏组织病理学的影响;同时设正常对照组及阳性对照组。结果：ＺＧＴ能显著降低大鼠血清转氨酶、ＭＤＡ、ＨＡ、ＰＣⅢ和ＬＮ,提高肝组织中ＳＯＤ的活性;明显减轻肝脏纤维增生程度。结论：ＺＧＴ对大 实验性肝纤维化有预防作用。     

慢性乙型肝炎肝组织糜酶活性与肝纤维化相关性研究

目的探讨慢性乙型肝炎患者肝组织糜酶活性与肝纤维化相关性。方法收集2009年1月至2012年3月延边大学附属医院消化科住院的慢性乙型肝炎患者46例,按肝脏病理肝纤维分级(S1～S4)进行分组,S1组10例,S2组15例,S3组15例,S4组6例;采用酶联免疫吸附试验(ELISA)检测肝组织中糜酶活性,采用胃酶酸解法测定肝组织胶原纤维及羟脯氨酸(HYP)含量,经Masson染色后直接测量胶原纤维面积与视野总面积比值,以该比值作为胶原纤维的相对含量。观察比较各组肝组织糜酶活性、胶原纤维含量和HYP含量。结果 S3组和S4组肝组织糜酶活性、肝脏胶原纤维含量及HYP含量均较S1组、S2组明显升高,差异有统计学意义(P<0.01),且纤维化程度重组(S3+S4)肝组织糜酶活性、胶原纤维含量及HYP含量明显高于纤维化轻组(S1+S2)(P<0.01);肝组织糜酶活性与肝组织胶原氨酸含量和HYP含量呈正相关,相关系数分别为0.685(P<0.05)和0.702(P<0.01)。结论慢性乙型肝炎患者肝组织中糜酶活性与肝组织胶原纤维含量和HYP含量呈正相关,可作为肝纤维化的评价指标之一。     

甘草酸二铵对二甲基亚硝胺诱导肝纤维化大鼠肝脏胶原增生沉积的影响

目的观察甘草酸二铵的抗肝纤维化作用。方法二甲基亚硝胺腹腔注射诱导大鼠肝纤维化模型,以甘草酸二铵7mg/100g体重灌胃治疗,每日1次,共4周,并以γ干扰素为对照药物。观察大鼠体重、肝脾重量等一般情况;肝脏苏木精-伊红染色与丽春红胶原染色,分级分期观察肝组织的炎性坏死与胶原纤维沉积变化;检测血清肝功能变化与肝组织羟脯氨酸含量。结果模型大鼠肝脏胶原纤维间隔形成,肝小叶与肝窦内胶原增生沉积明显;脾脏明显增大;血清ALT活性升高,肝组织羟脯氨酸含量上升。甘草酸二铵可显著降低模型大鼠肝组织羟脯氨酸含量,减轻肝脏内胶原纤维增生沉积,降低血清ALT活性。与γ干扰素比较作用无明显差异。结论甘草酸二铵有良好的抗二甲基亚硝胺诱导的大鼠肝纤维化效果,其作用机制可能与药物减轻肝脏炎症,抑制肝脏羟脯氨酸与胶原生成有关。     

排钱草总生物碱对实验性肝纤维化动物相关指标的作用

目的 观察排钱草总生物碱对实验性肝纤维化的作用效果及其作用机制。方法 采用四氯化碳（CC14）诱导肝纤维化模型,分别按阻断肝纤维化实验研究方法和治疗肝纤维化实验研究方法,从而清相关指标、肝脏羟脯氨酸含量和病理组织学等观察排钱草总生物碱的疗效,以秋水仙碱作为阳性对照药。结果 排钱草总生物碱能显著降低肝纤维化大鼠血清ALT、HA、γ-球蛋白及肝组织羟脯氨酸含量。肝脏病理组织学检查亦显示良好的作用效果。结论 排钱草总生物碱具有较好的抗肝纤维化作用。     

肝纤维化组织学量化诊断的实验研究

目的:探讨肝纤维化组织学量化诊断方法。方法:用二甲基亚硝胺(DMN)诱导SD大鼠肝纤维化模型,常规肝组织切片、Masson三色和苦味酸-天狼红染色,苦味酸固绿-天狼红染色比色测定,图像定量处理和半定量计分分析及羟脯氨酸(Hyp)含量测定,结果:切片胶原比色测定、图像定量处理和半定量计分分析均与肝纤维化程度变化相一致,与肝组织Hyp含量具有良好的相关性,r分别为0.94、0.89和0.88,P均<0.01。结论:切片胶原测定、图像定量处理和半定量计分可作为确定肝纤维化程度的量化指标。     

肥大细胞及蛋白酶激活受体-2在大鼠实验性肝纤维化中的变化

目的观察肝纤维化大鼠的肥大细胞(MC)数量变化和蛋白酶激活受体-2(PAR-2)在肝脏中表达及其与肝纤维化的关系。方法建立大鼠肝纤维化模型,以甲苯胺蓝染色显示MC,运用碱水解法测定肝脏羟脯氨酸含量,采用逆转录聚合酶链反应(RT-PCR)方法检测正常对照组及造模2、4、8、12周组大鼠肝组织中PAR-2 mRNA的表达,免疫组织化学方法检测PAR-2蛋白的表达及定位。结果正常对照组大鼠肝组织中MC数量极少,仅(2．5±1．0)个,主要沿肝脏汇管区分布；模型组MC数量：2周为(9．1±0．5)个,4周为(15．7±3．0)个,8周为(32．0±3．3)个。造模组2周与正常组比较,P=0．038,造模组组间比较,F=58．553,P＜0．01,差异均有统计学意义。MC在汇管区、中央静脉周围密集分布。随着造模时间的延长,肝脏羟脯氨酸的含量逐渐升高。PAR-2 mRNA检测结果,PAR- 2/β-肌动蛋白(β-actin)：正常对照组几乎不表达；造模2周肝组织可有少量PAR-2 mRNA表达,为0．15±0．01,4周为0．35±0．02,8周为0．80±0．02。PAR-2蛋白检测结果,阳性信号灰度：正常对照组为156．0±0．5；造模2周为162．5±1．3,4周为174．8±1．3,8周为185．7±2．1,12周为207．7±4．4,模型组与对照组比较和模型组组间比较,F=429．389,P＜<0．01,差异均有统计学意义。结论PAR-2 mRNA和蛋白的表达水平与MC数量、羟脯氨酸含量的增加一致,可能在肝纤维化的发生发展过程中起重要作用。     

Role of hepatic vitamin A and lipocyte distribution in experimental hepatic fibrosis

ABSTRACT— Lipocytes are the major site of hepatic vitamin A storage, and they have been demonstrated to lose their vitamin A content in the process of hepatic fibrosis. To investigate the relationship between hepatic vitamin A content and the degree of hepatic fibrosis, we measured levels of retinyl palmitate and retinol in the CCl₄-induced fibrotic liver using high-performance liquid chromatography. We estimated hepatic collagen content using a spectrophotometric analysis with sirius red, and also by measuring hydroxyproline levels. Lipocytes were detected by an immunoperoxidase method with anti-desmin antibody, and were counted morphometrically through a Texture Analyzing System. A significant negative correlation was observed between the level of retinyl palmitate and collagen content (r = –0.64) as well as the hydroxyproline level (r = –0.69) in the CCl₄-induced fibrotic liver. In the process of fibrosis, hepatic retinol levels were elevated in association with a decrease in retinyl palmitate. In particular in the early stage of fibrosis, lipocytes increased remarkably in number in fibrotic areas in spite of a decrease in total hepatic vitamin A. The present study suggests that an increase in hepatic retinol as well as a decrease in retinyl palmitate may facilitate the process of hepatic fibrosis produced by lipocytes.     

Antifibrotic effects of green tea on in vitro and in vivo models of liver fibrosis

AIM: To examine the protective effect of green tea extract (GT) on hepatic fibrosis in vitro and in vivo in dimethylnitrosamine (DMN)-induced rats. METHODS: HSC-T6, a rat hepatic stellate cell line, was used as an in vitro assay system. Cell proliferation,collagen content, and type 1 collagen expression were examined in activated HSC-T6 cells. Collagen was determined by estimating the hydroxyproline content.In rats with DMN-induced hepatic fibrosis, serum aspartate aminotransferase and alanine aminotransferase concentrations, liver hydroxyproline and lipid peroxides were determined. Pathologic changes were examined by hematoxylin & eosin staining.RESULTS: GT administration prevented the development of hepatic fibrosis in the rat model of DMN-induced liver fibrosis. These results were confirmed both by liver histology and by quantitative measurement of hepatic hydroxyproline content, a marker of liver collagen deposition. Accordingly, inhibition of proliferation, reduced collagen deposition, and type 1 collagen expression were observed in activated HSC-T6 cells following GT treatment. These results imply that GT reduced the proliferation of activated HSC and down regulated the collagen content and expression of collagen type 1, thereby ameliorating hepatic fibrosis. tea administration can effectively improve liver fibrosis caused by DMN, and may be used as a therapeutic option and preventive measure against hepatic fibrosis.     

扶正化瘀胶囊治疗日本血吸虫病肝纤维化的临床疗效观察

目的:探讨扶正化瘀胶囊治疗日本血吸虫病肝纤维化的疗效。方法:临床观察日本血吸虫病肝纤维化患者48例,随机分为试验组(基础护肝治疗联合扶正化瘀胶囊)和对照组(基础护肝治疗),每组24例,患者均在护肝及抗纤维化治疗前1个月给予吡喹酮驱虫治疗,治疗前后观察患者临床症状和体征的改善,肝功能、血清肝纤维化指标的复常以及影像学指标的改变。结果:治疗24周后,两组患者临床症状和体征与治疗前比较均有明显改善,尤其肝区不适症状的缓解,试验组优于对照组(P<0.05);两组患者治疗后血清丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、总胆红素(TBil)水平均有显著改善(P<0.05);试验组血清透明质酸(HA)﹑Ⅲ型前胶原(PCⅢ)水平在治疗12周、24周较治疗前均显著下降(P<0.05),下降幅度均显著优于对照组(P<0.05);治疗24周患者血清层粘连蛋白(LN)﹑Ⅳ型胶原(Ⅳ-C)含量与治疗前比较两组均有显著改善(P<0.05),且试验组改善优于对照组(P<0.05);治疗24周患者肝门静脉主干内径、脾脏厚度的改变,试验组明显优于治疗前(P<0.05),且与对照组比较差异具有显著性意义(P<0.05)。结论:扶正化瘀胶囊治疗日本血吸虫病肝纤维化可明显改善患者临床症状和体征,促进肝脏功能恢复,有效地减轻肝脏纤维化。     

A histological examination of liver fibrosis and tissue prolyl hydroxylase in various liver diseases

Prolyl hydroxylase (PH) is an enzyme acting in early stage of collagen synthesis. We have emphasized the significance of the measurement of serum PH (SIRPH) in relation to liver fibrosis mainly in patients with alcoholic liver disease (ALD). In this study, we determined the localization and positivity of PH by tissue PH stain method (Avidin Biotin Complex method) to clarify the differences in fibrosis between ALD (25 cases) and non-alcoholic liver diseases (non-ALD, 47 cases). Tissue PH was found to be positive in liver cells around Glisson's sheath in early stage of fibrosis, and then in liver cells left within septa and also in mesenchymal cells in the sinusoidal wall as fibrosis progressed. Although there were basically no marked differences between ALD and non-ALD, ALD tended to show stronger tissue PH positivity for a degree of fibrosis, PH positivity in parenchymal cells was especially remarkable around pericellular fibrosis in ALD. These results clearly reflected the important role of liver parenchymal cells in progression of fibrosis.     

The protective effects of SA3443, a novel cyclic disulfide, on chronic liver injuries in rats.

The effects of (4R)-hexahydro-7,7-dimethyl-6-oxo-1,2,5-dithiazocine-4- carboxylic acid (SA3443), a novel cyclic disulfide compound, on the development of chronic liver injury were studied in rats, using two types of models, carbon tetrachloride (CCl4)-induced chronic liver injury and heterologous serum (swine serum)-induced liver fibrosis. SA3443 (30-100 mg/kg, p.o.) significantly suppressed increases in serum transaminase and alkaline phosphatase activity induced by CCl4-treatment for 10 weeks. This compound also inhibited increases in hepatic lipids and hydroxyproline content in CCl4-treated rats. In the histopathological studies, treatment with SA3443 resulted in a decrease in the degree of hepatic necrosis, fibrosis and steatosis. On the other hand, 8-weeks treatment with swine serum revealed hepatic fibrosis without appearance of necrosis or fatty accumulation. In this model, SA3443 (30 mg/kg, p.o.) reduced the hepatic hydroxyproline level, and diminished the formation of connective tissue in the liver. These findings indicate that SA3443 protects the liver against chronic liver injuries induced by CCl4 and heterogeneous serum.     

Decreased collagen accumulation by a prolyl hydroxylase inhibitor in pig serum-induced fibrotic rat liver

Hepatic fibrosis was induced in rats by repeated i.p. injections of pig serum. The hepatic hydroxyproline content increased to 2.1 times the normal control level at 6 weeks and to 3.2 times at 10 weeks. When P-1894B, an inhibitor of prolyl hydroxylase, was administered, there was a dose-dependent inhibition of the increase to nearly normal control levels at 6 and 10 weeks. There was also by histology a dose-dependent reduction in the degree of hepatic fibrosis. Hepatocellular damage was minimal and its extent did not vary with the degree of fibrosis or the treatment. P-1894B dose dependently reduced the hydroxylation of peptidyl proline in the fibrotic liver. These data suggest that P-1894B inhibited hepatic fibrogenesis by direct action on collagen but not by protection against hepatocellular damage leading to collagen formation. A prolyl hydroxylase inhibitor may be a candidate for use in treatment of hepatic fibrosis.     

A selective ROCK inhibitor, Y27632, prevents dimethylnitrosamine-induced hepatic fibrosis in rats

BACKGROUND: p160ROCK is a direct Rho target which mediates Rho-induced assembly of focal adhesions and stress fibers. We previously reported that Rho signaling pathways are involved in the activation of hepatic stellate cells (HSC) in vitro. The aim of the present study was to test the hypothesis that an inhibitor specific for p160ROCK (Y27632) could prevent experimental hepatic fibrosis induced by dimethylnitrosamine (DMN) in rats. METHODS: Y27632 was given orally at 30 mg/kg daily for 4 weeks after the first injection of DMN. The degree of fibrosis was evaluated by image analysis and also by measurements of collagen and hydroxyproline content in the liver. The expression of alpha-smooth muscle actin (alpha-SMA) in the liver and in the primary cultured HSC was also evaluated. Semi-quantitative RT-PCR was performed to evaluate the expression of type I collagen mRNA in the liver. RESULTS: Y27632 treatment significantly decreased the occurrence of DMN-induced hepatic fibrosis and reduced the collagen and hydroxyproline content and alpha-SMA expression in the liver. The expression of alpha-SMA in HSC was also suppressed in vitro. CONCLUSIONS: These findings indicate that inhibitors of the Rho-ROCK pathway might be useful therapeutically in hepatic fibrosis.