水芹总酚酸抗四氯化碳所致肝纤维化作用的实验研究

目的探讨水芹总酚酸对四氯化碳所致大鼠慢性肝纤维化模型的保肝和抗肝纤维化作用及其可能机制。方法采用四氯化碳复制慢性肝损伤动物模型（40%四氯化碳橄榄油溶液,皮下注射,首次剂量按0.5ml/100g,以后按0.3ml/100g体质量,3天1次,共42d）。自造模之日开始,灌胃给药,1次/d,连续42d。末次给药后禁食24h,取大鼠血和肝脏,检测血清肝功能（ALT、AST、ALB、ALP、TP、ChE、LDH）指标、肝纤维化指标（HA、LN、Ⅲ-C、Ⅳ-C）和肝组织中SOD活力与MDA含量及免疫组化法测α-SMA、TGF-β1、TIMP-1的表达量和光镜下观察肝脏组织学变化。结果模型组大鼠有明显肝损伤和慢性纤维化表现;各给药组肝功能和肝纤维化均有不同程度的改善,病理组织学检查结果表明其肝脏病变程度均较模型组明显减轻。结论水芹总酚酸对慢性肝纤维化大鼠具有明显的保肝和抗纤维化作用,其机制可能与清除自由基和抗氧化作用以及抑制肝星状细胞的活化有关。     

软肝胶囊对四氯化碳所致慢性肝纤维化大鼠的肝保护作用

目的验证软肝胶囊对四氯化碳(CCl4)所致大鼠慢性肝纤维化模型的肝保护作用及抗纤维化作用。方法模型组及各给药组首次皮下注射40%CC145ml/kg,以后每3d注射1次40%CCl43ml/kg,正常对照组给予等容积橄榄油,持续9周,给药自造模之日开始。结果四氯化碳所致慢性肝纤维化大鼠有明显肝损伤及慢性纤维化的表现,软肝胶囊三个剂量组肝功能指标均有不同程度的改善;1.4,2.8g/kg组肝组织胶原蛋白含量较模型组均明显降低;0.7g/kg组HA的值较模型组显著降低;病理组织学检查结果表明其肝脏病变程度均较模型组明显减轻。结论软肝胶囊对慢性肝纤维化大鼠具有肝保护作用及抗肝纤维化作用。     

T1rho值对四氯化碳模型大鼠肝纤维化演变过程及严重程度的评估价值

目的探讨肝脏旋转坐标系下的自旋晶格弛豫时间(T1rho)值在四氯化碳(CCl4)建模大鼠肝纤维化进展及转归过程中的变化及对肝纤维化分期的价值。方法选取80只Sprague-Dawley大鼠采用随机数字表法分为3组,分别为CCl4组49只、恢复组20只、对照组11只,将大鼠分别标记行MRI基线扫描。CCl4组及恢复组大鼠经颈背部皮下注射CCl4进行建模,CCl4组于注药后4、6、8、10、12周末分别进行黑血T1rho扫描;恢复组大鼠于注药后4、6周末进行黑血T1rho扫描,第6周末扫描结束后停止注药,停止注药后1、2、4、6周末分别进行扫描;对照组于相同时间点注射等量玉米油,并于注射后第4、6、8、10、12周末分别进行黑血T1rho扫描。测量所有大鼠肝脏T1rho值,观察各组大鼠肝脏T1rho值随时间的变化。采用独立样本t检验比较各组大鼠相邻时间点肝脏T1rho值的差异。将实验鼠分为无肝纤维化组(S0)、轻度肝纤维化组(S1、2)及中重度肝纤维化组(S3、4),采用Kruskal-Wallis H检验分析3组间T1rho值间的差异,用受试者操作特征(ROC)曲线评估T1rho值对不同程度肝纤维化的诊断效能,采用Spearman检验评估T1rho值与肝纤维化严重程度的相关性。结果59只大鼠完成了整个实验,其中CCl4组28只、恢复组20只、对照组11只。CCl4组大鼠肝脏T1rho值随着注药时间的延长逐渐增加,8周末达到最大值,随后8~12周逐渐下降,除4周末与6周末、10周末与12周末大鼠肝脏的T1rho值差异无统计学意义(P值分别为0.112、0.487),其他相邻两个时间点大鼠肝脏T1rho值差异均有统计学意义(P均<0.05);恢复组大鼠肝脏T1rho值随着注药时间的延长逐渐上升,停止CCl4注射后肝脏T1rho值逐渐下降,相邻两个时间点大鼠肝脏T1rho值差异均有统计学意义(P均<0.05);对照组大鼠相邻时间点肝脏T1rho值差异均无统计学意义(P均>0.05)。无肝纤维化组(S0)、轻度肝纤维化组(S1、2)及中重度肝纤维化组(S3、4)大鼠分别为15、23、21只,T1rho值分别为[36.3(34.4,41.4)]、(47.2±8.4)、(48.8±9.0)ms,大鼠肝脏T1rho值随着肝纤维化程度的加重而加重,二者呈低度正相关(r=0.402,P=0.001);上述3组大鼠肝脏T1rho值间差异有统计学意义(P<0.01)。肝脏T1rho值鉴别无肝纤维化(S0)与有肝纤维化(S1~4)的ROC下面积为0.825(95%可信区间0.720~0.931),鉴别无或轻度肝纤维化(S0~2)与中重度肝纤维化(S3、4)的ROC下面积为0.668(95%可信区间0.540~0.796)。结论T1rho值对评估CCl4建模大鼠肝纤维化进展及转归有一定意义,对鉴别是否存在肝纤维化诊断价值中等,对鉴别轻度肝纤维化与中重度肝纤维化诊断价值较低。     

肝纤维化动物模型制备的常用方法及其MRI检测的研究进展

制备良好的动物模型是科研过程中的重要环节。制作肝纤维化动物模型的化学方法中最常用的主要有四氯化碳(CCl4)法和硫代乙酰胺(TAA)法,该方法造模简单、成模率高,物理方法中主要为胆管结扎法,该方法操作复杂、成模率较低,但对其他组织器官无毒性作用。在磁共振检测的研究中,弥散加权、灌注成像及波谱成像能够对中重度肝硬化作出分期,但对轻中度肝纤维化分期尚有明显不足,而弥散张量成像在轻中度的肝纤维化的诊断中有较大优势。现就制备动物肝纤维化模型的常用方法及该模型MRI检测的研究方法的进展进行综述。     

软肝合剂治疗慢性乙肝致肝纤维化疗效观察

动物实验证明,软肝合剂具有良好的保肝及预防肝纤维化作用[1]。我们将软肝合剂用于治疗慢性乙肝患者,并与常规治疗方法相比较,软肝合剂疗效较好。1　对象和方法1.1　对象　慢性乙肝51例,男43例,女8例。均符合1995年5月第5届全国传染病寄生虫病学术会议修订的诊断标准[2]。随机分为对照组31例,男26例,女5例;平均年龄30.4岁,平均病程5.3年。其中中度慢肝21例,重度慢肝10例。治疗组20例,男17例,女3例;平均年龄32.6岁,平均病程6.1年。其中中度慢肝14例,重度慢肝7例。两组性别、年龄、病程、病情具有可比性。1.2　治疗方法　对照组给予常规保肝及…     

自配软肝合剂治疗慢性乙肝致肝纤维化疗效观察

动物实验证明 ,软肝合剂具有良好的保肝及阻断和逆转肝纤维化向肝硬化发展的作用[1] 。因此 ,我院于 1998～ 1999年采用自制软肝合剂治疗慢性乙肝致肝纤维化 ,并与常规治疗组进行比较 ,前者效果较好。1　对象和方法1 1　对象　慢性乙肝 80例 ,随机分为软肝合剂组4 0例 ,男 34例 ,女 6例 ;平均年龄 32 6岁 ;平均病程 6 1年。中度慢肝 2 7例 ,重度慢肝 13例。常规治疗组 4 0例中 ,男 35例 ,女 5例 ;平均年龄 30 4岁 ;平均病程 5 7年。中度慢肝 2 8例 ,重度慢肝 12例。诊断及分型均符合 1995年 5月第五届全国传染病与寄生虫病学术会议修订的…     

肝纤维化的基因治疗

肝纤维化（1iverfibrosis）是肝脏因病毒、酒精、化学毒物、寄生虫等各种损害因子长期刺激的结果，并最终可能发展至肝硬化和肝功能衰竭。目前研究证明人类的肝纤维化是可逆的，而肝硬化是不可逆的，故针对肝纤维化的治疗显得尤为重要。但目前尚无成熟有效的抗肝纤维化药物问世，随着分子生物学及基因治疗技术的发展，肝纤维化的分子机制逐渐得以阐明，     

活动性肝纤维化大鼠模型中注射钆塞酸二钠后的钆沉积及肝纤维化

摘要目的评估钆塞酸二钠对活动性肝纤维化大鼠模型中纤维化的影响。材料与方法本研究已获动物实验委员会批准。在12周内腹腔注射四氯化碳(CCl4)诱导出肝纤维化。在CCl4的最终剂量后开始连续5d给予10mmol/kg的钆塞酸二钠(临床剂量为0.25mmol/kg)。使用3组SD大鼠进行实验。第1组的6只大鼠用钆塞酸二钠处理,第2组的9只大鼠用CCl4处理,第3组的9只同时使用钆塞酸二钠和CCl4处理。     

愈肝胶囊抗大鼠肝纤维化的实验研究

目的 探讨愈肝胶囊对大鼠实验性肝纤维化的治疗作用。方法 以四氯化碳制备大鼠肝纤维化模型,观察愈肝胶囊对肝纤维化大鼠肝脏病理变化,对肝功能指标ALT、TBIL、ALB、GLO、PT;肝纤维化指标HA、LN、PCⅢ、CⅣ、Hyp及免疫学指标CIM、CD8、CIM／CD8的作用来说明愈肝胶囊对肝纤维化大鼠的影响。结果 愈肝胶囊明显减轻肝纤维化大鼠肝脏的病理改变,改善其肝功能;调节T淋巴细胞;下调肝纤维化4项指标及羟脯氨酸;使肝纤维化程度显著下降,与乙肝宁颗粒组相比有显著性差异（P〈0．05-0．01）。结论 愈肝胶囊有显著降低大鼠实验性肝纤维化反应,增强机体抗肝纤维化损伤能力的作用。     

苦参素抗大鼠肝纤维化作用的实验研究

目的　研究苦参素对四氯化碳诱导的大鼠肝纤维化肝组织Smad基因表达的影响。方法　 90只SD大鼠分为 3组 ,正常对照组 (C组 )、模型组 (M组 )和苦参素干预组 (T组 )。以CCl4皮下注射法诱导大鼠肝纤维化 ,干预组大鼠在造模的同时给予苦参素注射液腹腔注射 ,正常对照组给予液体石蜡皮下注射和生理盐水腹腔注射 ,8周实验结束时处死动物。酶联免疫吸附法检测血清TGFβ1水平 ,HE和Masson染色观察大鼠肝组织病理学变化和胶原沉积 ,免疫组化法检测Smad3基因表达情况。结果　苦参素干预组大鼠血清TGFβ1水平下降 (32 .6 5± 8.71)vs(96 .37± 16 .5 4 )mg/L ,P <0 .0 5 ) ,肝组织组织学积分 (1.9± 0 .3vs3.6± 0 .8,P <0 .0 5 )和胶原面积 ((94 .4 1± 37.2 6 )vs(6 90 .86± 188.71) μm2 ,P <0 .0 5 )明显减少 ;Smad3蛋白表达阳性率减少 ((2 .33± 1.6 4 )vs(9.5 6± 1.34) % ,P <0 .0 5 )。结论　苦参素干预性治疗能够有效地减少大鼠肝组织胶原沉积 ,使其血清TGFβ1水平下降 ,并抑制Smad3基因表达 ,干扰TGFβ介导的肝纤维化信号转导     

NO在CCl4致大鼠肝纤维化中的氧化应激作用

目的探讨一氧化氮(NO)在CCl4致大鼠肝纤维化中的作用。方法Wistar大鼠34只,雄性,体重200~220g,用CCl4制造大鼠肝纤维化模型,另设正常对照组大鼠6只。HE染色、Massons染色,观察肝组织病理学改变;检测血清NO,肝组织丙二醛(MDA)、超氧化物歧化酶(SOD)。结果肝纤维化模型组大鼠第7周达到2期或3期肝纤维化,汇管区周围纤维隔形成并相互连接,部分纤维隔深入小叶内,小叶结构保留或紊乱,但无肝硬化,伴有中度脂肪变性及肝细胞空泡变性;血清NO水平明显增高(152·8±12·30μmol/Lvs.11·03±1·95μmol/L,P<0·05),肝组织MDA含量明显增高(2·11±0·26nmol/mgprotvs.0·73±0·13nmol/mgprot,P<0·05),SOD活性明显降低(44·96±5·36nU/mgprotvs.105·73±13·5nU/mgprot,P<0·05)。结论随着大鼠肝纤维化的形成,大鼠血清NO水平随之增高,进一步导致肝细胞炎症及损伤,促进了肝纤维化的形成。     

Diffusion tensor imaging of liver fibrosis in an experimental model

Purpose

To characterize changes in diffusion properties of liver using diffusion tensor imaging (DTI) in an experimental model of liver fibrosis.

Materials and Methods

Liver fibrosis was induced in Sprague–Dawley rats (n = 12) by repetitive dosing of carbon tetrachloride (CCl₄). The animals were examined with a respiratory‐gated single‐shot spin‐echo echo‐planar DTI protocol at 7 T before, 2 weeks after, and 4 weeks after CCl₄ insult. Apparent diffusion coefficient (ADC), directional diffusivities (ADC_// and ADC_?), and fractional anisotropy (FA) were measured. Liver histology was performed with hematoxylin‐eosin staining and Masson's trichrome staining.

Results

Significant decrease (P < 0.01) in ADC was found at 2 weeks (0.86 ± 0.09 × 10^?3 mm²/s) and 4 weeks (0.74 ± 0.09 × 10^?3 mm²/s) following CCl₄ insult, as compared with that before insult (0.97 ± 0.08 × 10^?3 mm²/s). Meanwhile, FA at 2 weeks (0.18 ± 0.03) after CCl₄ insult was significantly lower (P < 0.01) than that before insult (0.26 ± 0.05), and subsequently normalized at 4 weeks (0.26 ± 0.07) after the insult. Histology showed collagen deposition, presence of intracellular fat vacuoles, and cell necrosis/apoptosis in livers with CCl₄ insult.

Conclusion

DTI detected the progressive changes in water diffusivities and diffusion anisotropy of liver tissue in this liver fibrosis model. ADC and FA are potentially valuable in detecting liver fibrosis at early stages and monitoring its progression. Future human studies are warranted to further verify the applicability of DTI in characterizing liver fibrosis and to determine its role in clinical settings. J. Magn. Reson. Imaging 2010;32:1141–1148. © 2010 Wiley‐Liss, Inc.

     

Liver fibrosis: an intravoxel incoherent motion (IVIM) study

Purpose:

To characterize longitudinal changes in molecular water diffusion, blood microcirculation, and their contributions to the apparent diffusion changes using intravoxel incoherent motion (IVIM) analysis in an experimental mouse model of liver fibrosis.

Materials and Methods:

Liver fibrosis was induced in male adult C57BL/6N mice (22–25 g; n = 12) by repetitive dosing of carbon tetrachloride (CCl₄). The respiratory‐gated diffusion‐weighted (DW) images were acquired using single‐shot spin‐echo EPI (SE‐EPI) with 8 b‐values and single diffusion gradient direction. True diffusion coefficient (D_true), blood pseudodiffusion coefficient (D_pseudo), and perfusion fraction (P_fraction) were measured. Diffusion tensor imaging (DTI) was also performed for comparison. Histology was performed with hematoxylin‐eosin and Masson's trichrome staining.

Results:

A significant decrease in D_true was found at 2 weeks and 4 weeks following CCl₄ insult, as compared with that before insult. Similarly, D_pseudo values before injury was significantly higher than those at 2 weeks and 4 weeks after CCl₄ insult. Meanwhile, P_fraction values showed no significant differences over different timepoints. For DTI, significant decrease in ADC was observed following CCl₄ administration. Fractional anisotropy at 2 weeks after CCl₄ insult was significantly lower than that before insult, and subsequently normalized at 4 weeks after the insult. Liver histology showed collagen deposition, the presence of intracellular fat vacuoles, and cell necrosis/apoptosis in livers with CCl₄ insult.

Conclusion:

Both molecular water diffusion and blood microcirculation contribute to the alteration in apparent diffusion changes in liver fibrosis. Reduction in D_true and D_pseudo values resulted from diffusion and perfusion changes, respectively, during the progression of liver fibrosis. IVIM analysis may serve as valuable and robust tool in detecting and characterizing liver fibrosis at early stages, monitoring its progression in a noninvasive manner. J. Magn. Reson. Imaging 2012;36:159–167. © 2012 Wiley Periodicals, Inc.     

二咖啡酰奎宁酸抗肝纤维化及脂质过氧化作用的实验研究

目的 :观察二咖啡酰奎宁酸抗复合因素致大鼠肝纤维化及脂质过氧化作用。方法 :采用复合因素复制大鼠肝纤维化模型 ,以血清层连蛋白 (LN)、透明质酸 (HA)、丙二醛 (MDA)、一氧化氮 (NO)含量、肝组织光镜、电镜下形态学改变为观察内容 ,探讨二咖啡酰奎宁酸的治疗作用。结果 :二咖啡酰奎宁酸能够显著降低血清LN ,HA ,MDA ,NO含量 ,明显改善肝组织炎症及纤维化状态。结论 :二咖啡酰奎宁酸具有一定的抗肝纤维化及脂质过氧化作用     

盐酸博莱霉素致大鼠肺纤维化动物模型HRCT的评价

目的 评价高分辨率CT(HRCT)对盐酸博莱霉素致大鼠肺纤维化动物模型的诊断及鉴别诊断价值,探讨能应用于放射影像学研究的肺纤维化动物模型。方法 雄性SD大鼠60只,其中50只经气管滴注盐酸博莱霉素,10只经气管滴注等体积的生理盐水。28天后平行观察大鼠HRCT征象,行HRCT-病理对照研究。结果 经气管一次性滴注盐酸博莱霉素的大鼠均出现了纤维化病变,HRCT可见肺实变、结节影、血管支气管束异常、胸膜增厚、交界面不规则、磨玻璃样密度影等,部分可见蜂窝肺,病理切片对照显示肺组织纤维性增生。结论 HRCT方法适于诊断活体动物肺纤维化,该方法方便、准确、可靠,具有良好的可重复性。     

Effect of disease progression on liver apparent diffusion coefficient and T2 values in a murine model of hepatic fibrosis at 11.7 Tesla MRI

Purpose:

To evaluate the effects of hepatic fibrosis on ADC and T₂ values of ex vivo murine liver specimens imaged using 11.7 Tesla (T) MRI.

Materials and Methods:

This animal study was IACUC approved. Seventeen male, C57BL/6 mice were divided into control (n = 2) and experimental groups (n = 15), the latter fed a 3, 5‐dicarbethoxy‐1, 4‐dihydrocollidine (DDC) supplemented diet, inducing hepatic fibrosis. Ex vivo liver specimens were imaged using an 11.7T MRI scanner. Spin‐echo pulsed field gradient and multi‐echo spin‐echo acquisitions were used to generate parametric ADC and T₂ maps, respectively. Degrees of fibrosis were determined by the evaluation of a pathologist as well as digital image analysis. Scatterplot graphs comparing ADC and T₂ to degrees of fibrosis were generated and correlation coefficients were calculated.

Results:

Strong correlation was found between degrees of hepatic fibrosis and ADC with higher degrees of fibrosis associated with lower hepatic ADC values. Moderate correlation between hepatic fibrosis and T₂ values was seen with higher degrees of fibrosis associated with lower T₂ values.

Conclusion:

Inverse relationships between degrees of fibrosis and both ADC and T₂ are seen, highlighting the utility of these parameters in the ongoing development of an MRI methodology to quantify hepatic fibrosis. J. Magn. Reson. Imaging 2012;35:140‐146. © 2011 Wiley Periodicals, Inc.     

Evaluation of hepatic fibrosis with portal pressure gradient in rats

MRI has the potential of providing a noninvasive assessment of liver pathology. This work introduces a portal pressure gradient (PPG) model derived from fluid mechanics, where the PPG is proportional to the average velocity and inversely proportional to the vessel area in the upper part of portal vein. Using a phase‐contrast spoiled gradient echo sequence, the PPG model was verified in a phantom study and was tested in an animal study using 35 rats with various degrees of hepatic fibrosis induced by carbon tetrachloride (CCl₄). Histological examination was conducted to determine the severity of hepatic fibrosis. The fibrosis score monotonically increased with the duration of CCl₄ treatment. The PPG was highly correlated with nonzero fibrosis scores (r² = 0.90, P < 0.05). There was a significant difference between control and cirrhosis groups (P < 0.0006, α < 0.0018). The difference between control and fibrosis (noncirrhosis) groups (P < 0.002, α < 0.006) was also significant. Without the administration of any contrast agent, the MRI‐PPG approach shows promise as a noninvasive means of evaluating liver fibrosis. Magn Reson Med, 2009. © 2009 Wiley‐Liss, Inc.     

MR elastography for noninvasive assessment of hepatic fibrosis: experience from a tertiary center in Asia

Purpose:

To determine the sensitivity and specificity of MR elastography (MRE) in the staging of hepatic fibrosis (HF) using histopathology as the reference standard in an Asian population.

Materials and Methods:

MRE was performed on 55 patients with chronic liver diseases or biliary diseases and on 5 living related liver donors (48 men and 12 women; mean age, 55.7 years). MRE was performed with modified, phase‐contrast, gradient‐echo sequences, and the mean stiffness values were measured on the elastograms in kilopascals(kPa). Receiver operating characteristic curve analysis was performed to determine the cutoff value and accuracy of MRE for staging HF. Histopathologic staging of HF according to the METAVIR scoring system served as the reference.

Results:

Liver stiffness increased systematically along with the fibrosis stage. With a shear stiffness cutoff value of 3.05 kPa, the predicted sensitivity and specificity for differentiating significant liver fibrosis (≥ F2) from mild fibrosis (F1) were 89.7% and 87.1%, respectively. In addition, MRE was able to discriminate between patients with severe fibrosis (F3) and those with liver cirrhosis (sensitivity, 100%; specificity, 92.2%), with a shear stiffness cutoff value of 5.32 kPa.

Conclusion:

MRE could be a promising, noninvasive technique with excellent diagnostic accuracy for detecting significant HF and liver cirrhosis. J. Magn. Reson. Imaging 2011. © 2011 Wiley Periodicals, Inc.     

Quantitative assessment of hepatic fibrosis in an animal model with magnetic resonance elastography.

Chronic liver disease is a world-wide problem that causes progressive hepatic fibrosis as a hallmark of progressive injury. At present, the gold standard for diagnosing hepatic fibrosis is liver biopsy, which is an invasive method with many limitations, including questionable accuracy and risks of complications. MR elastography (MRE), a phase-contrast MRI technique for quantitatively assessing the mechanical properties of soft tissues, is a potential noninvasive diagnostic method to assess hepatic fibrosis. In this work, MRE was evaluated as a quantitative method to assess the in vivo mechanical properties of the liver tissues in a knockout animal model of liver fibrosis. This work demonstrates that the shear stiffness of liver tissue increases systematically with the extent of hepatic fibrosis, as measured by histology. A linear correlation between liver stiffness and fibrosis extent was well-defined in this animal model. An additional finding of the study was that fat infiltration, commonly present in chronic liver disease, does not significantly correlate with liver stiffness at each fibrosis stage and thus does not appear to interfere with the ability of MRE to assess fibrosis extent. In conclusion, MRE has the potential not only for assessing liver stiffness, but also for monitoring potential therapies for hepatic fibrosis.