肝纤维化的表观遗传学研究进展

肝纤维化是常见的慢性进行性肝病,是慢性肝病发展到肝硬化阶段的必经阶段,却有逆转的可能性。肝纤维化的中心环节是肝星状细胞的激活。关于肝星状细胞的激活,除了经典的肝纤维化通路,不断有新的通路和机制出现,包括自噬、内质网应激、氧化应激、胆固醇代谢和表观遗传,这些都揭示了肝星状细胞的活化机制。表观遗传包括DNA甲基化、组蛋白修饰和调节性非编码RNAs,这些机制也参与调节肝星状细胞活化和肝纤维化发生,对表观遗传和肝纤维化治疗之间的关系研究具有重要意义。     

活化的肝星状细胞RBP-J可诱导性基因敲除小鼠的构建及应用

目的:利用Cre-loxp可诱导系统构建活化的肝星状细胞RBP-J可诱导性基因敲除小鼠模型,以实现肝纤维化过程中活化的肝星状细胞Notch信号通路的特异性阻断。方法:将Sm22αCre ERT2和RBP-Jflox/flox转基因小鼠杂交,得到F1小鼠,将繁殖的F1小鼠继续进行交配,得到F2小鼠,通过PCR鉴定出基因型为Sm22αCre ERT2,RBP-Jflox/flox的小鼠。通过腹腔注射四氯化碳建立肝纤维化模型,检测四氯化碳注射不同时间段后肝脏纤维化进展情况和肝星状细胞活化过程中Sm22α的表达情况。注射他莫昔芬诱导活化肝星状细胞中RBP-J基因的特异性敲除。分选肝星状细胞,q-PCR和Western Blot检测活化肝星状细胞中Notch下游靶基因Hes1、Hey1表达情况以验证敲除效果。结果:通过连续交配我们获得了Sm22αCre ERT2,RBP-Jflox/flox小鼠。四氯化碳腹腔注射4周即可诱导肝脏发生较为明显的纤维化,同时肝星状细胞活化并逐渐高表达Sm22α。给予他莫昔芬诱导Cre重组酶发挥作用后,敲除了活化的肝星状细胞中的RBP-J基因,其下游基因Hes1、Hey1表达降低70%以上,阻断了Notch信号通路。结论:我们成功构建了RBP-J可诱导性条件性基因敲除小鼠模型,实现了小鼠肝纤维化过程中活化肝星状细胞Notch信号通路的阻断,为深入研究Notch信号通路在肝纤维化中的作用和机制奠定了坚实的基础。     

肝星状细胞中表皮形态发生素表达调控机制及其作用研究

肝星状细胞是肝脏中重要的间质细胞,是肝细胞外基质的主要来源.表皮形态发生素(epimorphin、EPM、syntaxin2)在肝脏发育、再生及癌变过程中发挥了重要的作用,目前其表达变化的调控机制及对肝星状细胞的作用还未有报道.通过对肝组织标本进行检测,发现肝纤维化过程中肝星状细胞表达EPM上调.从表观遗传学的角度对EPM表达变化调控机制进行研究,发现DNA去甲基化促进了EPM的表达.为了研究EPM对肝星状细胞的可能的调节作用,将EPM表达质粒转染肝星状细胞,之后检测了EPM对肝星状细胞增殖及迁移能力的变化.结果证明EPM能够促进肝星状细胞的增殖与迁移.本研究发现,激活的肝星状细胞高表达EPM可能是由于DNA去甲基化引起的,同时,高表达的EPM能够促进肝星状细胞的增殖与迁移,进而促进肝纤维化进展.     

肝纤维化发生发展及逆转过程中肝巨噬细胞亚群分类

肝纤维化是由持续性损伤修复反应引起的,导致肝组织内细胞外基质异常沉积,进一步引发肝脏结构和肝功能异常改变的一种病理过程.已有大量研究表明,肝纤维化在去除损伤因素后是可以逆转的.肝星状细胞作为主要的效应细胞,合成和分泌各种胶原和细胞外基质,一直被认为是肝纤维化发生发展的中心环节.最近的研究发现,巨噬细胞作为主要的调节细胞,能同时调节肝星状细胞的功能和基质胶原的降解,促进肝纤维化的形成.而在肝纤维化逆转过程中,促使活化的肝星状细胞凋亡和纤维胶原的降解,促进肝纤维化的逆转.目前已有研究表明,巨噬细胞亚群在肝纤维化发生发展及逆转中具有双向调控作用,但是对于动物模型体内还没有系统的研究巨噬细胞亚群的分类.本文对巨噬细胞亚群的分类研究做一个全面的综述,对肝脏巨噬细胞在肝纤维化中分子机制的进一步研究具有一定的参考价值和借鉴意义.     

肝纤维化发病机制中细胞和分子机制的研究进展

关于肝纤维化形成的复杂的细胞和分子联系已经有了相当多的研究进展。最近的数据表明,纤维化进程的终止和纤维分解途径的恢复可以逆转晚期肝纤维化甚至肝硬化。因此,需要更好地阐明参与肝纤维化的细胞和分子机制。HSC(肝星状细胞)的激活是肝纤维化发生的中心事件,此外还有产生基质的其他细胞来源,包括肝门区的成纤维细胞,纤维细胞和骨髓来源的肌纤维母细胞。这些细胞与其邻近细胞通过多种联系聚集产生纤维疤痕并造成持续性损伤。阐明不同类型的细胞的相互作用,揭示细胞因子对这些细胞的影响,理清活化HSC基因表达的调控,将有助于我们探索新的肝纤维化治疗靶点。此外,不同的病因有不同的致病途径,弄清这一点有助于针对特异性疾病治疗方法的发现。本文概述了肝纤维化的细胞和分子机制的最新研究进展,可能为未来治疗方法带来新的突破。     

细胞自噬与肝纤维化

自噬是细胞内一种进化保守的依赖溶酶体的蛋白降解途径,存在于大多数真核细胞中,细胞自噬可以为应激状态(如炎症、肿瘤、毒物等)的细胞提供能量。已有研究表明,肝纤维化的发病机制中涉及肝星状细胞的活化,伴随着细胞内脂质小滴的消耗,当肝细胞经历自噬,溶酶体的胞内基质将下降。研究还发现肝星状细胞的活化过程中伴随着其自噬水平的升高。本文将就自噬在肝纤维化中的作用做一综述。     

细胞珠蛋白对肝星状细胞氧化损伤的保护作用

为揭示细胞珠蛋白对肝星状细胞氧化损伤的保护作用及相关机制,通过siRNA干扰内源性细胞珠蛋白基因,利用重组细胞珠蛋白作用于完全活化的人肝星状细胞系LX-2及大鼠原代肝星状细胞,并在LX-2细胞内过表达细胞珠蛋白,考察在过氧化氢及铁过载两种不同作用机制的氧化反应模型中细胞的增殖性及细胞内超氧化物水平。结果表明内源性细胞珠蛋白对于两种氧化反应导致的肝星状细胞损伤都具有显著性的保护作用,证明其在活化肝星状细胞内的表达上调是其应对氧化应激的保护性措施;重组细胞珠蛋白不仅能保护完全活化的LX-2细胞免受氧化应激损伤,并且能抑制未完全活化的原代肝星状细胞过度增殖以及保护其被过度损伤;重组细胞珠蛋白对细胞内的活性氧清除效果不理想,可能与其进出细胞缺乏相应的主动运输机制有关。进一步在LX-2细胞内过表达细胞珠蛋白对无论是铁过载或是过氧化氢引起的氧化反应均能发挥较好的保护性作用。为加速肝纤维化药物新靶点开发提供了理论依据。     

Hippo-YAP/TAZ与Notch信号通路在肝纤维化中的作用

病毒性或代谢性慢性肝病引起的肝纤维化是全球健康的一大挑战。Hippo-YAP/TAZ通路与Notch信号通路在肝纤维化进展中发挥了至关重要的作用。经典的Hippo通路核心激酶级联在大量外界信号的作用下，通过磷酸化使转录共激活因子YAP/TAZ失活，从而调控细胞的生长与增殖以及干细胞再生、肿瘤形成等过程。Notch信号通路通过调控脂质代谢、IR、氧化应激、自噬、炎症与纤维化等方面参与肝病的发生与发展。该文就Hippo-YAP/TAZ与Notch信号通路在肝纤维化过程中对肝星状细胞、巨噬细胞、肝脂质代谢等方面的影响及两条通路的交互作用进行了详细地综述。概括了2条通路在肝纤维化中的研究现状，总结分析了交互作用研究中存在的问题和未来的方向，旨在为肝纤维化的防治提供新的靶点和理论依据。     

牛磺酸上调基因1(TUG1)对肝纤维化的作用及其机制*

目的:探讨牛磺酸上调基因1(TUG1)在肝纤维化中的作用机制。方法:按照文献建立TGF-β1(5 ng/ml)刺激的活化肝星状细胞模型和经典的1%DMN(1 ml/kg/d)致大鼠肝纤维化模型,将肝纤维化大鼠和活化肝星状细胞(HSC)均分为模型对照组、阴性对照组(沉默TUG1阴性对照)、siRNA干扰组(TUG1基因沉默组)。实验结束后利用苏木精-伊红(HE)染色检测大鼠肝脏组织病理变化;采用逆转录-聚合酶链反应(RT-PCR)法、蛋白免疫印记(Western blot)分别测定大鼠肝组织及活化肝星状细胞中α-平滑肌肌动蛋白(α-SMA)、TUG1、I型胶原蛋白(collagenI)、基质金属蛋白酶-2(MMP-2)、金属蛋白酶组织抑制因子(TIMP-1)、Smad2、Smad3表达水平。结果:肝组织病理学检查显示,沉默TUG1能够明显缓解肝脏纤维化病理改变,Western blot结果显示,沉默TUG1能够显著降低大鼠肝组织和活化肝星状细胞中TUG1、α-SMA、collagenI、MMP-2、TIMP-1、Smad2、Smad3基因与蛋白表达水平(P＜0.05)。与模型对照组相比,阴性对照组的TUG1、α-SMA的蛋白与基因水平明显升高(P＜0.05)。与模型对照组和阴性对照组相比,siRNA干扰组中TUG1, α-SMA, collagenI, MMP-2, TIMP-1, Smad2 and Smad3的蛋白和基因水平显著降低(P＜0.05),而在模型对照组和阴性对照组中TUG1, α-SMA, collagenI, MMP-2, TIMP-1, Smad2 and Smad3的蛋白和基因表达水平之间差异无显著性。结论:TUG1在肝纤维化组织和活化的肝星状细胞中显著上调,沉默TUG1可能通过抑制转化生长因子-β1(TGF-β1)/Smad信号通路改善1%DMN致大鼠肝纤维化病理损伤,降低活化肝星状细胞中纤维化相关蛋白水平,发挥抗肝纤维化的作用。     

姜黄素对大鼠肝星状细胞表达受体型蛋白酪氨酸磷酸酶的影响的实验研究

目的:本研究通过观察受体型蛋白酪氨酸磷酸酶(PTPRJ)在大鼠肝星状细胞中的表达及姜黄素对其的影响,探讨以PTPRJ为靶点,姜黄素对抗肝纤维化的分子机制.方法:四氯化碳(CCL4)皮下注射诱导大鼠肝纤维化模型,并采用Percoll梯度离心法分离培养原代大鼠肝星状细胞,采用western-blot、RT-PCR法分别测定PTPRJ、其mRNA在肝组织及肝星状细胞的表达变化及姜黄素对其表达的影响.结果:western-blot、RT-PCR检测结果显示:肝组织和肝星状细胞中PTPRJ表达呈阳性,并随着肝纤维化进展,其表达逐渐降低;姜黄素作用后,肝组织和肝星状细胞PTPRJ表达明显增多.结论:姜黄素可显著增强肝星状细胞PTPRJ的表达,从而改善或逆转肝纤维化进展.     

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。     

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。     

The collagenolytic effects of the traditional Chinese medicine preparation,Han-Dan-Gan-Le,contribute to reversal of chemical-induced liver fibrosis in rats

Han-Dan-Gan-Le (HDGL), a Chinese herb preparation composed of Stephaniat tetrandra, Salvia miltorrhiza, Radix paeoniae, Astragalus membranaceus, and Ginkgo biloba, has been used to treat human liver fibrosis. This study was designed to examine the therapeutic effect of HDGL on chemical-induced liver fibrosis in adult Wistar rats. Liver fibrosis was produced in rats by carbon tetrachloride (1.2 ml CCl(4)/kg, 2 times/week, after an initial dose of 5.0 ml CCl(4)/kg, sc), plus a diet of 20% fat, 0.05% cholesterol (continuous) and 30% alcohol in the drinking water ad libitum (every other day) for 8 weeks. HDGL (0.5 and 1.0 g/kg, ig, daily for 6 weeks) was administered to rats 72 hrs after the last dose of CCl(4) to examine its therapeutic effects on chemical-induced liver fibrosis. Upon pathological examination, the HDGL treatment had significantly reversed chemical-induced liver fibrosis and other hepatic lesions. Hepatic collagen accumulation induced by CCl(4) was markedly reduced by HDGL treatment, as evidenced by hepatic collagen content and by immunohistochemical analysis of type-I collagen in liver. HDGL appeared to stimulate the collagenolytic process in the liver, as a 30-50% increase in urinary excretion of hydroxyproline was observed with HDGL treatment as compared to rats only given CCl(4). In conclusion, HDGL can effectively reverse chemically induced liver fibrosis, and this appears to be due, at least in part, to the stimulation of hepatic collagenolysis, resulting in a resolution of hepatic fibrosis.     

MiR-542-3p controls hepatic stellate cell activation and fibrosis via targeting BMP-7

There has been an increasing number of studies about microRNAs as key regulators in the development of hepatic fibrosis. Here, we demonstrate that miR-542-3p can promote hepatic fibrosis by downregulating the expression of bone morphogenetic protein 7 (BMP-7), which is known to antagonize transforming growth factor β1 (TGFβ1)-mediated fibrogenesis effect. The expression of miR-542-3p is increased in activated hepatic stellate cells (HSCs). Downregulation of MiR-542-3p by antisense inhibitors can inhibit HSCs activation markers, including α-smooth muscle actin (α-SMA) and collagen as well as TGFβ signaling pathways. MiR-542-3p was significantly upregulated in carbon tetrachloride (CCl₄)-induced hepatic fibrosis in mice, and downregulation of miR-542-3p by lentivirus could prevent the development of hepatic fibrosis. In addition, miR-542-3p can directly bind to the 3′-untranslated region of BMP-7 mRNA, indicating that its profibrotic effect appears to be caused by its inhibition of BMP-7. Our results suggest that downregulation of miR-542-3p prevents liver fibrosis both in vitro and in vivo, highlighting its potential as a novel biomarker or therapeutic target for hepatic fibrosis.     

The osteopontin level in liver, adipose tissue and serum is correlated with fibrosis in patients with alcoholic liver disease

Background

Osteopontin (OPN) plays an important role in the progression of chronic liver diseases. We aimed to quantify the liver, adipose tissue and serum levels of OPN in heavy alcohol drinkers and to compare them with the histological severity of hepatic inflammation and fibrosis.

Methodology/Principal Findings

OPN was evaluated in the serum of a retrospective and prospective group of 109 and 95 heavy alcohol drinkers, respectively, in the liver of 34 patients from the retrospective group, and in the liver and adipose tissue from an additional group of 38 heavy alcohol drinkers. Serum levels of OPN increased slightly with hepatic inflammation and progressively with the severity of hepatic fibrosis. Hepatic OPN expression correlated with hepatic inflammation, fibrosis, TGFβ expression, neutrophils accumulation and with the serum OPN level. Interestingly, adipose tissue OPN expression also correlated with hepatic fibrosis even after 7 days of alcohol abstinence. The elevated serum OPN level was an independent risk factor in estimating significant (F≥2) fibrosis in a model combining alkaline phosphatase, albumin, hemoglobin, OPN and FibroMeter® levels. OPN had an area under the receiving operator curve that estimated significant fibrosis of 0.89 and 0.88 in the retrospective and prospective groups, respectively. OPN, Hyaluronate (AUROC: 0.88), total Cytokeratin 18 (AUROC: 0.83) and FibroMeter® (AUROC: 0.90) estimated significance to the same extent in the retrospective group. Finally, the serum OPN levels also correlated with hepatic fibrosis and estimated significant (F≥2) fibrosis in 86 patients with chronic hepatitis C, which suggested that its elevated level could be a general response to chronic liver injury.

Conclusion/Significance

OPN increased in the liver, adipose tissue and serum with liver fibrosis in alcoholic patients. Further, OPN is a new relevant biomarker for significant liver fibrosis. OPN could thus be an important actor in the pathogenesis of this chronic liver disease.     

Human amniotic epithelial cell transplantation induces markers of alternative macrophage activation and reduces established hepatic fibrosis

Chronic hepatic inflammation from multiple etiologies leads to a fibrogenic response that can progress to cirrhosis and liver failure. Transplantation of human amniotic epithelial cells (hAEC) from term delivered placenta has been shown to decrease mild to moderate hepatic fibrosis in a murine model. To model advanced human liver disease and assess the efficacy of hAEC therapy, we transplanted hAEC in mice with advanced hepatic fibrosis. Immunocompetent C57BL/6 mice were administered carbon tetrachloride (CCl(4)) twice weekly resulting in bridging fibrosis by 12 weeks. hAEC (2 × 10(6)) were infused via the tail vein at week 8 or weeks 8 and 10 (single and double dose, respectively). Human cells were detected in mouse liver four weeks after transplantation showing hAEC engraftment. CCl(4) treated mice receiving single or double hAEC doses showed a significant but similar decrease in liver fibrosis area associated with decreased activation of collagen-producing hepatic stellate cells and decreased hepatic protein levels of the pro-fibrogenic cytokine, transforming growth factor-beta1. CCl(4) administration caused hepatic T cell infiltration that decreased significantly following hAEC transplantation. Hepatic macrophages play a crucial role in both fibrogenesis and fibrosis resolution. Mice exposed to CCl(4) demonstrated increased numbers of hepatic macrophages compared to normal mice; the number of macrophages decreased significantly in CCl(4) treated mice given hAEC. These mice had significantly lower hepatic protein levels of the chemokine monocyte chemoattractant protein-1 than mice given CCl(4) alone. Alternatively activated M2 macrophages are associated with fibrosis resolution. CCl(4) treated mice given hAEC showed increased expression of genes associated with M2 macrophages including YM-1, IL-10 and CD206. We provide novel data showing that hAEC transplantation induces a wound healing M2 macrophage phenotype associated with reduction of established hepatic fibrosis that justifies further investigation of this potential cell-based therapy for advanced hepatic fibrosis.     

黑木耳多糖提取物对肝纤维化大鼠肝功能及MPO, MDA, TGF-beta的影响

目的：探究黑木耳多糖提取物对肝纤维化大鼠血清髓过氧化物酶(MPO)、丙二醛(MDA)及转化生长因子beta(TGF-beta)的影响及 肝功能的保护作用。方法：将健康的Wistar大鼠30 只随机分为3 组：对照组、模型组、黑木耳多糖提取物组,利用胆总管结扎术制 备肝纤维化动物模型,术后分别用生理盐水0.1 mL/(kg·d)、黑木耳多糖提取物0.1 mg/(kg·d)灌胃1 次/d。21 d后腹主动脉采血,测 肝功能指标,取肝组织测MDA、MPO、TGF-beta,Masson 染色。结果：肝纤维化动物模型成功制备。黑木耳多糖提取物组总胆红素 (STB)、谷氨酸转移酶(ALT)、天门冬氨酸转移酶(AST)、MPO、MDA较模型组明显降低,TGF-茁含量降低。Masson 染色可见：模型 组肝小叶结构破坏,较对照组汇管区可见大面积胶原纤维的形成;黑木耳多糖提取物组肝细胞轻度水肿、肝索排列欠整齐,肝汇 管区及各部位胶原纤维形成量略少。结论：黑木耳多糖提取物可减轻胆总管结扎大鼠脂质过氧化损伤,MPO 含量使TGF-beta因子 释放减少,从而减少胶原纤维形成,减轻或延缓大鼠肝纤维化。     

Hepatic fibrogenesis using chronic arsenic ingestion: studies in a murine model

Chronic oral arsenic (As) ingestion has been alleged to cause hepatic fibrosis, non-cirrhotic portal fibrosis and cirrhosis of the liver. The present study was aimed to investigate if hepatic fibrogenesis and non-cirrhotic portal fibrosis (NCPF) is caused by arsenic. A significant increase in the hepatic protein and collagen was seen compared with controls; hepatic 4-hydroxyproline levels, indicative of fibrogenesis, were increased 4-14 folds with different dosages of arsenic compared to the controls. Hepatocellular necrosis and inflammation were negligible to mild in all the groups. None of the animals developed significant splenomegaly or features of non-cirrhotic portal hypertension. The results suggest that (i) prolonged oral arsenic ingestion in mice leads to significant hepatic fibrogenesis and collagen synthesis with minimal hepato-cellular injury; (ii) arsenic ingestion alone is unlikely to cause non-cirrhotic portal fibrosis or cirrhosis of liver. This murine model of arsenic feeding could be used for the evaluation of new antifibrotic agents for the liver.     

Effectiveness of Losartan-Loaded Hyaluronic Acid (HA) Micelles for the Reduction of Advanced Hepatic Fibrosis in C3H/HeN Mice Model

Advanced hepatic fibrosis therapy using drug-delivering nanoparticles is a relatively unexplored area. Angiotensin type 1 (AT1) receptor blockers such as losartan can be delivered to hepatic stellate cells (HSC), blocking their activation and thereby reducing fibrosis progression in the liver. In our study, we analyzed the possibility of utilizing drug-loaded vehicles such as hyaluronic acid (HA) micelles carrying losartan to attenuate HSC activation. Losartan, which exhibits inherent lipophilicity, was loaded into the hydrophobic core of HA micelles with a 19.5% drug loading efficiency. An advanced liver fibrosis model was developed using C3H/HeN mice subjected to 20 weeks of prolonged TAA/ethanol weight-adapted treatment. The cytocompatibility and cell uptake profile of losartan-HA micelles were studied in murine fibroblast cells (NIH3T3), human hepatic stellate cells (hHSC) and FL83B cells (hepatocyte cell line). The ability of these nanoparticles to attenuate HSC activation was studied in activated HSC cells based on alpha smooth muscle actin (α-sma) expression. Mice treated with oral losartan or losartan-HA micelles were analyzed for serum enzyme levels (ALT/AST, CK and LDH) and collagen deposition (hydroxyproline levels) in the liver. The accumulation of HA micelles was observed in fibrotic livers, which suggests increased delivery of losartan compared to normal livers and specific uptake by HSC. Active reduction of α-sma was observed in hHSC and the liver sections of losartan-HA micelle-treated mice. The serum enzyme levels and collagen deposition of losartan-HA micelle-treated mice was reduced significantly compared to the oral losartan group. Losartan-HA micelles demonstrated significant attenuation of hepatic fibrosis via an HSC-targeting mechanism in our in vitro and in vivo studies. These nanoparticles can be considered as an alternative therapy for liver fibrosis.