动物模型在间充质干细胞治疗慢性肝病临床前研究的应用

慢性肝病进一步发展为肝纤维化并导致肝硬化,此时除了器官移植外没有更为有效的治疗方法。然而器官缺乏的问题迫使人们寻找更为有效的治疗策略。间充质干细胞(mesenchymal stem cells,MSCs)有着免疫调节、分化成肝细胞、促进原位肝细胞再生和抑制肝星状细胞的激活等能力,所以利用MSCs开展细胞移植治疗具有非常广阔的前景。本文通过综述各类人的肝病动物模型和运用这些动物模型开展MSCs细胞移植治疗肝病的临床前研究的进展和意义,将为今后在MSCs开展临床治疗的广泛应用中提供安全性和有效性评价的科学依据。     

治疗肝纤维化的蛋白质药物研究进展

肝纤维化是肝脏出现瘢痕组织,瘢痕尽管在肝脏的位置有异,但是均一致表现为细胞外基质的增生和性质转变,尤其在疾病的早期。蛋白质药物在治疗疾病方面有许多优势,我们将用于治疗肝纤维化的蛋白质药物做出总结,为新药研究提供思路。     

四氯化碳诱导的小鼠肝纤维化模型中SDF-1/αCXCR4的表达及其意义

目的检测四氯化碳（CCl4）诱导的肝纤维化小鼠模型肝脏SDF-1/αCXCR4的表达,评价SDF-1/αCXCR4轴与肝纤维化的关系,为研究肝纤维化肝损伤发生及损伤修复机制研究提供基础。方法选用6周龄雌性纯系C57小鼠,采用40%的CCl4/橄榄油溶液腹腔注射,剂量为1 mL/kg,每周2次,共4周,制成肝纤维化模型,取肝纤维化及正常对照组小鼠肝脏标本,采用RT-PCR及免疫组化检测SDF-1α的表达,采用RT-PCR及Western检测CXCR4受体的表达。结果与对照组相比,SDF-1α及CXCR4在肝纤维化模型小鼠肝脏组织中的表达较对照组明显上调,差异具有统计学意义（P〈0.05）。结论肝纤维小鼠肝组织的SDF-1/αCXCR4受体表达上调,为研究肝纤维化肝损伤机制及干细胞移植治疗提供理论基础。     

肝纤维化的实验动物模型

本文讨论了建立肝纤维化动物模型的技术与存在的问题,以大鼠为主要动物讨论了目前常用的几种造模方法、每种造模方法的致病机理、造模途径、效果,同时比较了不同造模方法的优缺点及各自的用途,提出了肝纤维化动物模型领域的研究方向。     

鳖甲在抗肝纤维化和抗肝癌作用中的研究进展

鳖甲性寒味咸,归肝肾经,能滋阴潜阳,善于软坚散结。张仲景创立的经典方剂鳖甲煎丸方中以鳖甲为主药,具有行气活血、祛湿化痰、软坚消癥的功效。近年来鳖甲及其复方鳖甲煎丸在临床上广泛应用于肝硬化及肝癌的治疗,取得了较好的疗效。在作用机制研究中发现鳖甲的抗肝纤维化作用可能与其对机体炎症环节的影响有关,同时对细胞外基质(ECM)和肝星状细胞(HSC)具有调控作用;在抗肝癌研究中,发现单味鳖甲和鳖甲煎丸可能通过增强小鼠的免疫功能、肝脏保护作用,发挥抑制肝癌细胞生长的作用。本研究对鳖甲及其复方鳖甲煎丸在肝纤维化和肝癌的临床治疗应用,以及实验研究进展做一综述,以期为研究鳖甲在抗肝纤维化和抗肝癌作用中的机制研究,及肝纤维化及肝癌的治疗和预防提供参考。     

SOCS-3在肝纤维化中的作用及中药干预的研究进展

慢性肝病是全世界发病率及死亡率最高的疾病之一。肝纤维化是各种慢性肝病进展为肝硬化的必经阶段。大量研究证实,肝纤维化的病理过程可以逆转,但目前尚未被攻克,因此,寻找有效治疗肝纤维化的方法十分必要。近些年来研究表明,SOCS-3在肝纤维化不同阶段扮演不同角色,其可通过影响细胞增殖、细胞周期、细胞因子及信号转导等干预肝纤维化,可能成为治疗肝纤维化的潜在靶标。同时,很多中药可通过影响SOCS-3来发挥改善肝纤维化的作用。本文就SOCS-3在肝纤维化中的作用及中药干预的研究进行分析综述,为进一步研究治疗肝纤维化的有效方法提供参考。     

骨髓间充质干细胞抗肝纤维化的研究进展

肝纤维化及其终末病变肝硬化已严重危害全球人类健康,虽然慢性肝病的治疗手段和抗肝纤维化药物的研究已取得了很大进展,肝移植依然是最有效的治疗方案,但器官的紧缺却是一个现实问题。目前寻找有效的干预手段进行抗肝纤维化治疗已越来越受到大家的关注。近些年,大量基础及临床研究均证实在一定条件下利用骨髓间充质干细胞(MSCs)可以抑制肝星状细胞活化诱导其凋亡,实现肝纤维化逆转。随着干细胞技术的快速发展,基于骨髓间充质干细胞(MSCs)的细胞疗法在肝纤维化治疗领域的研究与应用已成为一个充满生命力的新方向。本文将对肝纤维化及基于MSCs的治疗机制进展及其应用进行综述。     

差异蛋白质组学在肝纤维化疾病中的应用

差异蛋白质组学作为一种新兴的蛋白质研究技术,逐渐在肝纤维化(HF)疾病的研究中广泛应用。该文主要对差异蛋白组学的理论基础、在肝纤维化疾病的诊治和研究技术上的应用以及目前存在的问题等方面作一简要综述,从而阐明在肝纤维化疾病中差异蛋白组学现阶段的研究现状,旨在为探讨如何更全面完整地研究肝纤维化提供参考。随着科学技术及科研条件的不断完善,差异蛋白组学的多种技术手段应用于肝纤维化疾病将具有更广阔的研究前景和发展空间。     

非酒精性脂肪肝炎动物模型的研究概况

随着全球代谢综合征患病率不断上升,非酒精性脂肪肝病(NAFLD)患病率也急剧上升,已是全球最常见的慢性肝疾病,包括单纯性脂肪肝、非酒精性脂肪性肝炎(NASH)、晚期纤维化、肝硬化和肝细胞癌.而NASH是由单纯性脂肪肝发展至肝纤维化、肝硬化和肝癌的重要阶段,因此,预防和治疗NASH已成为一个研究焦点.动物模型是研究疾病的...     

肝巨噬细胞与肝星状细胞交互作用对 肝纤维化发生与逆转的影响

肝纤维化是由各种病因所导致的肝脏病理性反应,是发展成肝硬化甚至肝癌的必经途径.以往研究发现,肝纤维化甚至是肝硬化早期都可以通过一定的干预治疗抑制与逆转病情,该过程有多种肝实质以及非实质细胞参与,肝星状细胞(hepatic stellate cell,HSC)与肝巨噬细胞是肝纤维化进程中关键的细胞类型.HSCs是肝纤维化...     

超声导入疗法治疗慢性乙型肝炎肝纤维化临床研究

目的：探讨超声导入疗法对乙型肝炎肝纤维化患者进行治疗的临床效果。方法：选择符合诊断标准的慢性乙型肝炎肝纤维化患者52例,随机分为试验组和对照组,各26例。对照组患者给予基本保肝治疗,试验组在对照组的基础上加用黄芪注射液进行超声导入,3个月为1个疗程。观察两组患者治疗前后症状、体征、血清肝纤维化指标、肝功能变化及影像学指标。结果：两组患者症状、体征均有不同程度的改善,差异无统计学意义（P〉0．05）;试验组血清肝纤维化指标明显改善,与对照组比较,差异有统计学意义（P〈0．05）;肝功能及影像学指标的改善更明显（P〈0．01）。结论：超声导入疗法对慢性乙型肝炎肝纤维化具有改善肝功能,减少肝细胞外基质的增生与沉积的效用,能够减轻或延缓肝纤维化的进展。     

Decorin-TGFβ axis in hepatic fibrosis and cirrhosis

Hepatic fibrosis and cirrhosis are worldwide health care problems, especially in regions with a high rate of hepatitis infection. As these diseases affect a major part of the human population, the search for antifibrotic therapies has a high priority in medical research. Transforming growth factor β1 (TGF-β1) is one of the most powerful profibrotic cytokines. Thus, blocking TGF-β1 activity by natural inhibitors represents a valid and logical strategy to combat hepatic fibrosis. One of the natural inhibitors of TGF-β1 is decorin, a small leucine-rich proteoglycan that binds with high affinity to this cytokine and prevents its interaction with pro-fibrotic receptors. Recent evidence has shown that decorin has a protective role in liver fibrogenesis insofar as its genetic ablation in mice leads to enhanced matrix deposition, impaired matrix degradation, and "activation" of hepatic stellate cells, the main producers of fibrotic tissue. Moreover, TGF-β1 exerts a stronger effect when functional decorin is absent. These data provide robust genetic evidence for a direct role of endogenous decorin in preventing and retarding hepatic fibrosis. Thus, boosting the endogenous production of decorin or systemic delivery of recombinant decorin could represent an additional therapeutic modality against hepatic fibrosis.     

超声造影对肝硬化和肝纤维化的应用价值

目的：评估超声造影对于肝硬化和肝纤维化的I临床应用价值。方法：选取2011年5月至2013年5月在我院接受诊治的慢性病毒性肝炎患者32例,其中肝纤维化24例,肝硬化12例,均经过超声造影下的肝功能检测和肝穿刺病理证实。另选取10例到我院经检查身体健康的志愿者作为对照组,无任何肝脏病史。采用实时灰阶造影,经肘静脉注射造影SonoVue,记录测定造影剂进入肝静脉到达时间（HVAT）、肝动脉到达时间（HAAT）及肝动静脉渡越时间（HAVTT）。结果：肝纤维化组超声造影剂到达肝静脉的时间较对照组明显延长,肝硬化组较对照组和肝纤维化组的超声造影剂到达肝静脉的时间要明显缩短,差异符合统计学意义（P〈0．05）;造影剂到达肝动脉的时间组间比较无统计学差异;肝硬化组肝动静脉渡越时间（HAVTT）较对照组时间明显缩短,组间比较具有统计学差异（P〈0．05）;HAVTT随着肝纤维化程度的升高而缩短,中、重度肝纤维化患者的时间缩短较明显,差异具有统计学意义（P〈0．05）。结论：超声造影在肝硬化和肝纤维化诊断上具有一定的临床诊断价值,尤其是对于诊断肝纤维化程度具有重要的参考价值,可以更好地指导临床治疗,但是对于轻度的肝纤维化的诊断的准确性上有些缺陷。     

Alpha-lipoic acid inhibits hepatic PAI-1 expression and fibrosis by inhibiting the TGF-β signaling pathway

Accumulating evidence suggests that plasminogen activator inhibitor (PAI)-1 plays an important role in the development of hepatic fibrosis via its involvement in extracellular matrix remodeling. We previously reported that alpha-lipoic acid (ALA), a naturally occurring thiol antioxidant, prevents hepatic steatosis by inhibiting the expression of sterol regulatory element binding protein-1c. The aim of the present study was to determine whether ALA prevents hepatic PAI-1 expression and fibrosis through the inhibition of multiple TGF-β-mediated molecular mediators. We investigated whether ALA inhibited the development of hepatic fibrosis in mice following bile duct ligation (BDL), an established animal model of liver fibrosis. We found that ALA markedly inhibited BDL-induced hepatic fibrosis and PAI-1 expression. We also found that ALA attenuated TGF-β-stimulated PAI-1 mRNA expression, and inhibited PAI-1 promoter activity in liver cells; this effect was mediated by Smads and the JNK and ERK pathways. The results of the present study indicate that ALA inhibits hepatic PAI-1 expression through inhibition of TGF-β-mediated molecular mediators, including Smad3, AP1, and Sp1, and prevents the development of BDL-induced hepatic fibrosis. These findings suggest that ALA may have a clinical application in preventing the development and progression of hepatic fibrosis.     

Hepatic fibrosis--role of hepatic stellate cell activation

Hepatic fibrosis is a reversible wound healing response characterized by accumulation of extracellular matrix (ECM), or "scar," that follows chronic but not self-limited liver disease. The ECM components in fibrotic liver are similar regardless of the underlying cause. Activation of hepatic stellate cells is the central event in hepatic fibrosis. These perisinusoidal cells orchestrate an array of changes including degradation of the normal ECM of liver, deposition of scar molecules, vascular and organ contraction, and release of cytokines. Not only is hepatic fibrosis reversible, but it is also increasingly clear that cirrhosis may be reversible as well. The exact stage at which fibrosis/cirrhosis becomes truly irreversible is not known. Antifibrotic therapies will soon be a clinical reality. Emerging therapies will be targeted to those patients with reversible disease. The paradigm of stellate cell activation provides an important framework for defining therapeutic targets.     

Targeting secreted cytokine BMP9 gates the attenuation of hepatic fibrosis

Liver fibrosis is overly exuberant wound healing that leads to portal hypertension or liver cirrhosis. Recent studies have demonstrated the functions of bone morphogenetic protein 9 (BMP9) in liver fibrosis, and thus, targeting liver-specific BMP9 abnormalities will become an attractive approach for developing therapeutics to treat liver fibrosis. Here, we reveal that BMP9 serves as a valuable serum diagnostic indicator and efficient therapeutic target to attenuate liver fibrogenesis. Our analysis of biopsies from liver fibrotic patients revealed that higher BMP9 levels accompanied advanced stages of liver fibrosis. In mouse models, recombinant Bmp9 overexpression accelerated liver fibrosis, and adenovirus-mediated Bmp9 knockdown attenuated liver fibrogenesis. Intriguingly, BMP9 directly stimulated hepatic stellate cell activation via the SMAD signaling pathway to enhance hepatic fibrosis. Moreover, an inhibitory monoclonal antibody targeting Bmp9 was efficacious in treatment of mice with liver fibrosis. These observations delineate a novel model in which BMP9 directly drives SMAD/ID1 signaling in hepatic stellate cells, which modulates liver fibrogenesis development. Moreover, the findings unveil a promising surrogate biomarker for the diagnosis of hepatic fibrosis, thereby representing an efficient “BMP9 neutralization” approach in alleviating hepatic fibrosis.     

血清转化生长因子beta-1 在大鼠放射性肝纤维化中的动态表达 及其相关性研

目的:肝癌的放射治疗可导致放射性肝损伤(RILD)、甚至肝纤维化及肝硬化等并发症的发生,因此寻找较佳的血清标记物对放射性肝纤维化的无创诊断及监测具有重要意义。本文通过建立放射性肝纤维化大鼠模型,检测血清转化生长因子β1(TGF-β1)的动态表达,从而探讨其与放射性肝纤维化严重程度的相关性及其作为血清标记物的诊断价值。方法:雄性SD大鼠40只,随机分为模型组(30只)和对照组(10只)。除对照组外,模型组大鼠右半肝均接受单次6MV X线25Gy照射,于照射2月、4月、6月后,随机抽取10只,酶联免疫吸附法(ELISA)检测血清TGF-β1的表达,同时将大鼠肝组织进行HE染色,观察肝组织病理变化及大鼠肝纤维化程度,将后者与血清TGF-β1值进行相关性分析。结果:在照射第2月、4月、6月后,模型组大鼠血清TGF-β1值(分别为551.03±69.00 ng/L、645.31±109.29 ng/L、737.89±118.11 ng/L)逐渐升高,均明显高于对照组(451.71±51.12 ng/L,P<0.05)。通过相关性分析表明,大鼠血清TGF-β1值与肝纤维化程度正相关(r=0.82,P<0.01)。结论:在放射性肝纤维化发生发展中,TGF-β1随着肝纤维化严重程度增加,其表达亦升高。本研究为放射性肝纤维化严重程度的监测提供了一种新的无创、操作简单的手段,为后续TGF-β1作为血清标记物应用于放射性肝纤维化的临床研究奠定了理论基础。     

Thrombin cleavage of osteopontin controls activation of hepatic stellate cells and is essential for liver fibrogenesis

Liver biopsy is the current reliable way of evaluating liver fibrosis. However, no specific sera biomarker could be applied in clinical diagnosis. As the pivotal role of osteopontin (OPN) reported in numerous liver diseases, thrombin-cleaved OPN (Thr-OPN) exposes an integrin-binding motif that promoted biological functions. Herein, we investigated the potential of Thr-OPN in liver fibrosis. Using patient samples, mouse models and hepatic stellate cells (HSCs), we analyzed the involvement of Thr-OPN in liver fibrosis. The result showed that, first, Thr-OPN level was significantly higher in patients with liver cirrhosis than that in patients with chronic hepatitis B and healthy controls. Thr-OPN level was positively correlated with liver fibrosis degree in clinical samples. Then in mouse models, it showed a similar correlation between hepatic Thr-OPN levels and liver fibrosis degree. Thr-OPN peptides exacerbated liver fibrosis in OPN-deficient mice, whereas the neutralization of Thr-OPN alleviated liver fibrosis in wild-type mice. Furthermore, when compared with full-length OPN (FL-OPN), Thr-OPN exhibited a greater ability to promote HSC activation, proliferation, and migration via mitogen-activated protein (MAP) kinase and nuclear factor (NF)-κB pathways. In conclusion, Thr-OPN, not FL-OPN, was critically involved in the exacerbation of liver fibrosis by α9 and α4 integrins via MAP kinase and NF-κB signaling pathway, thus representing a novel diagnostic biomarker and treatment target for liver cirrhosis.     

Animal models for the study of liver fibrosis: new insights from knockout mouse models

Fibrosis arises as part of a would-healing response that maintains organ structure and integrity following tissue damage but also contributes to a variety of human pathologies such as liver fibrosis. Liver fibrosis is an abnormal response of the liver to persistent injury with the excessive accumulation of collagenous extracellular matrices. Currently there is no effective treatment, and many patients end up with a progressive form of the disease, eventually requiring a liver transplant. The clarification of mechanisms underlying pathogenesis of liver fibrosis and the development of effective therapy are of clinical importance. Experimental animal models, in particular targeted gene knockouts (loss of function) in mice, have become a powerful resource to address the molecular mechanisms or significance of the targeted gene in hepatic functions and diseases. This review will focus on the recent advances in knowledge obtained from genetically engineered mice that provide novel insights into the pathophysiology of liver fibrosis.