SIRT3与非酒精性脂肪性肝病肝细胞脂肪变性关系的临床研究

背景：非酒精性脂肪性肝病（NAFLD）是指在多种因素的共同作用下,肝细胞逐渐发生脂肪变性的临床病理过程。SIRT3与棕色脂肪的产热有关,肝细胞内可检测到其表达。目的：通过检测NAFLD患者肝活检标本中SIRT3、AMP活化蛋白激酶（AMPK）、乙酰辅酶A羧化酶1（ACC1）和同醇调节元件结合蛋白-1（SREBP—1）的表达,探讨SIRT3与NAFLD肝细胞脂肪变性的关系。方法：从体检人员和部分行肝胆外科手术者中纳入56例NAFLD患者,12例健康体检者作为对照组。受试者先以超声检查初步诊断脂肪肝程度,然后行肝穿刺活检,确定组织病理学肝细胞脂肪变性程度,以免疫组化方法检测SIRT3、AMPK、ACC1、SREBP-1表达。结果：超声诊断的脂肪肝分级与组织病理学肝细胞脂肪变性程度一致。脂肪肝患者肝细胞内SIRT3和AMPK表达量显著低于对照组（P〈0．05）,并随脂肪肝程度的加重而减低;ACC1和SREBP．1表达量显著高于对照组（P〈0．05）,并随脂肪肝程度的加重而增加。结论：SIRT3与NAFLD的肝细胞脂肪变性呈负相关．其表达降低可能通过下调AMPK表达,使脂肪合成基因ACC1和SREBP-1表达增加,导致肝细胞发生脂肪变性。     

内质网分子伴侣糖调节蛋白94在非酒精性脂肪肝中的表达及意义

目的:检测非酒精性脂肪肝患者肝组织中糖调节蛋白94的表达情况,探讨非酒精性脂肪肝病(nonalcoholic fattyliver disease,NAFLD)的发病机制.方法:应用半定量逆转录聚合酶链反应(RTPCR)法和免疫组化法检测脂肪肝组和正常对照组肝组织中糖调节蛋白94基因和蛋白表达,HE染色观察肝脏脂肪变性、炎性活动和坏死,并进行相关分析.结果:非酒精性脂肪肝组脂肪变性、炎症评分及坏死评分较正常对照组明显增加,差异具有统计学意义(3.89±0.32 VS 0.20±0.4;3.67±0.49 VS 0:0.44±0.51 VS 0,P＜0.01或0.05).与正常组比较,脂肪肝患者肝组织糖调节蛋白94mRNA和蛋白表达明显增强(0.86±0.02 VS 0.37±0.03;0.34±0.01 VS 0.31±0.01,均P＜0.01).糖调节蛋白94 mRNA和蛋白表达与脂肪变性、炎症评分及坏死评分明显呈正相关(P＜0.01).结论:非酒精脂肪肝引起的肝脏损害可能与糖调节蛋白94表达有关.     

非酒精性脂肪肝的研究新进展

非酒精性脂肪肝(NAFLD)是一种无过量饮酒史，而以肝实质细胞脂肪变性和脂肪贮积为特征的临床病理综合征。疾病谱随病程的进展表现不一，包括单纯脂肪肝、脂肪性肝炎、脂肪性肝纤维化和脂肪性肝硬化等4个病理过程。美国成年人3％-23％患有非酒精性脂肪肝。通过肝脏显像、尸检或肝活检发现70％肥胖病人有肝细胞脂肪变性，其中20％出现脂肪性肝炎，     

非酒精性脂肪肝自噬基因Beclin-1与核因子NF-κBp65表达研究

目的探讨自噬基因Beclin-1在高脂饮食诱导的非酒精性脂肪性肝病(NAFLD)大鼠模型的表达及对NAFLD发病机制的影响。方法(1)选取24只成年雄性SD大鼠随机分为两组:NAFLD模型组(高脂饲料饲养)12只,正常组12只(普通饲料饲养)。连续饲养12周后处死两组大鼠,采取心脏取血法,检测血清生化学指标ALT、AST、TG、CHOL及低密度脂蛋白(LDL-C)。(2)将大鼠肝脏组织取出,取少许行病理切片,采用HE染色,光镜下观察肝脏组织病理变化,进一步评估肝脏脂肪变性程度及造模是否成功;免疫组化染色,确定NF-κBp65表达。(3)应用Western blot方法检测NAFLD造模成功后的大鼠肝组织中自噬基因(Beclin-1)蛋白的表达水平。结果(1)成功建立了大鼠NAFLD模型,血生化指标ALT、AST、CHOL、TG及LDL-C水平与对照组同期相比均明显升高(P=0.0422,0.0336,0.0405,0.0411,0.0433)。(2)Western blot图像结果显示,与对照组比较,模型组肝组织Beclin-1蛋白表达明显升高(P=0.0411)。(3)正常组大鼠肝小叶结构清晰完整,肝索呈放射状分布于中央静脉周围;模型组大鼠肝细胞内可见脂滴,NF-κBp65阳性,评分为44.01±0.22;免疫组化结果显示,NF-κB集中沉积在汇管区肝细胞胞浆,低倍镜视野下平均占细胞总数55.1 5±1.01%。结论自噬基因Beclin-1与NF-κBp65在NAFLD模型肝组织中表达水平升高并可能促进NAFLD发生发展过程。     

高脂饮食性非酒精性脂肪性肝病大鼠肝脏PPAR-γ表达增强

目的　探讨过氧化物酶体增值物活化受体γ(PPAR-γ)及其亚型在高脂饮食所致非酒精性脂肪性肝病(NAFLD)大鼠肝脏的表达及其意义。方法　模型组SD大鼠给予高脂肪高胆固醇饮食饲养 ,分批于实验第 8、12、2 6、2 4周处死 ,同期设普通饮食饲养大鼠作对照。免疫组织化学和RT-PCR分别检测大鼠肝脏PPAR-γ的表达。结果　模型组大鼠第 8周呈现单纯性脂肪肝 ,第 12～ 2 4周从脂肪性肝炎进展为脂肪性肝炎伴肝纤维化。免疫组织化学和RT PCR显示 ,随着造模时间延长 ,肝脏PPAR-γ的表达逐渐增强。模型组肝脏PPAR-γ1mRNA表达于第 2 4周达到高峰 (与对照组相比升高 3 .5倍 ,P <0 .0 1) ,PPAR-γ2 mRNA表达于造模第 16周时达高峰 (较对照组升高 5 .8倍 ,P <0 .0 1)。相关分析显示 ,仅PPAR-γ2 mRNA与肝脂变程度之间关系密切 (r =0 .89,P <0 .0 5 )。结论　持续 2 4周的高脂饮食可以成功复制大鼠NAFLD模型 ,模型大鼠肝脏PPAR-γ表达增强 ,NAFLD大鼠肝细胞可能部分具有脂肪细胞的特征 ,即脂肪变的肝细胞发生成脂性改变     

高脂饮食诱导非酒精性脂肪性肝病小鼠模型肝脏microRNA表达谱的变化分析

目的研究microRNA(miRNA)-384的表达对高脂饮食诱导非酒精性脂肪性肝病(NAFLD)小鼠肝脂肪变性的影响。方法将30只雄性C57BL/6J小鼠适应性喂养7 d,然后随机分成2组,每组15只。对照组中的15只小鼠给予普通饮食,模型组15只小鼠给予HFD,喂养8周后获取肝组织。进行HE和尼罗红染色观察肝组织病理改变。采用微阵序列测序肝组织的全基因组miRNA表达谱,PCR法测定miRNA-384的相对表达水平。计量资料2组间比较采用t检验。结果对照组小鼠肝脏呈红色,边缘锐利,肝小叶结构清晰,肝细胞无脂肪变性;模型组小鼠肝脏呈黄色,边缘变钝,肝细胞明显肿胀,细胞质内可见大量脂肪空泡,脂滴相互融合导致细胞核偏位。与正常小鼠相比,在NAFLD小鼠的肝组织中发现了12个上调和18个下调的miRNA。筛查了对照组和模型组间的一些差异表达的miRNA,获得同样的聚类分析图。在这8个明显改变的miRNA中,miRNA-384具有显著的倍数变化。结论 miRNA-384表达的上调与肝脏脂肪变性有着密切关系,但是其作用机制仍需进一步研究。     

Fas/FasL、Bcl-2/Bax、Caspase-8在大鼠非酒精性脂肪性肝病中的作用机制

目的:探讨肝细胞凋亡及其相关因素Fas/FasL、Bcl-2/Bax蛋白及Caspase-8 mRNA的表达在大鼠非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)中的作用.方法:采用改良高脂饮食建立大鼠NAFLD模型,以正常饮食设立对照组.HE染色观察肝脏脂肪变、炎症活动和纤维化程度,采用流式细胞仪法测肝细胞凋亡百分数;免疫组织化学法检测Fas、FasL、Bcl-2及Bax在肝组织中表达情况;实时荧光定量PCR法检测肝脏Caspase-8 mRNA的表达.结果:NAFLD模型组脂肪变性明显,纤维化和炎症活动度记分均显著高于正常对照组.流式细胞仪检测显示,与对照组比较,实验组大鼠肝细胞凋亡百分数增加,随造模时间延长凋亡率增加更明显(均P<0.01);免疫组织化学染色显示随着肝脏脂肪变加重,Fas、FasL蛋白染色加深,阳性细胞数增加;Bcl-2、Bax在对照组的表达均为散在弱阳性,实验组4、8、12wk阳性细胞数渐增加,并在脂肪变明显的部位着色深且随着脂肪肝的进展,Bcl-2/Bax比率进行性下降.实时荧光定量PCR法显示Caspase-8 mRNA表达量在高脂组中显著高于对照组(均P<0.01),且随肝脏脂肪变及炎症加重呈进行性上升.结论:在NAFLD发生过程中,肝细胞凋亡促进NAFLD大鼠病情进展;Fas、FasL、Caspase-8 mRNA相关调控蛋白的活化是引起NAFLD脂肪变性、炎症及纤维化的重要因素.细胞凋亡调节蛋白Bax、Bcl-2表达上调,二者表达的相对比例发生异常,这可能是NAFLD中肝细胞发生凋亡的重要原因之一.     

非酒精性脂肪性肝病肝纤维化诊治研究进展

非酒精性脂肪肝（NAFLD）是指除外长期大量饮酒和其他损伤肝脏因素所引起的以肝脏脂肪沉积为主要表现的临床综合征,患者肝脏脂肪代谢功能出现明显障碍,使得大量脂肪类物质蓄积于肝细胞,导致肝细胞发生脂肪变性,从单纯性非酒精性脂肪肝（NAFL）发展到非酒精性脂肪性肝炎（NASH）,最终发展为肝纤维化、肝硬化和终末期肝病,甚至肝癌。近年来由于发病率逐渐升高引起人们的重视。     

非酒精性脂肪肝疾病发生机制研究新进展

非酒精性脂肪肝疾病(NAFLD)是一种无过量饮酒史的以肝实质细胞脂肪变性和脂肪贮积为特征的临床病理综合征,疾病谱随病程的进展而表现不一,包括单纯性脂肪肝、非酒精性脂肪性肝炎(NASH)、肝硬化甚至肝细胞癌〔1〕。NAFLD是西方     

非酒精性脂肪性肝病大鼠肝脏纤溶酶原激活物及其抑制物mRNA表达

目的　探讨非酒精性脂肪性肝病 (NAFLD)大鼠肝脏组织型纤溶酶原激活物 (t PA )及纤溶酶原激活物抑制物 1(PAI 1)基因表达及其意义。方法　高脂饮食建立SD大鼠NAFLD模型 ,分批于造模第 8、12、16、2 4周处死 ,同期设普通饮食喂养大鼠作对照。通过H E染色和苦味酸VG染色观察肝组织学改变 ,应用RT PCR对肝脏t PA和PAI 1mRNA的表达进行相对定量分析。结果　模型组大鼠于实验 8、12、2 4周分别形成单纯性脂肪肝、脂肪性肝炎以及脂肪性肝炎并肝纤维化。与对照组相比 ,模型组大鼠肝脏PAI 1mRNA表达随造模时间延长而增强 ,于实验 2 4周达高峰( 1.0 2± 0 .11比 0 .5 1± 0 .0 9,P <0 .0 1) ,并与其肝脂肪变及肝组织学损伤程度呈正相关 (r分别为 0 .492和 0 .3 72 ,P分别 <0 .0 1和 <0 .0 5 )。肝脏t PAmRNA表达随造模时间延长而逐渐减少 ,于实验 2 4周降至最低 ( 0 .89± 0 .11比 1.62± 0 .10 ,P <0 .0 1) ,但其仅与肝组织学损伤程度总积分呈负相关 (r =-0 .3 68,P <0 .0 5 )。结论　高脂饮食大鼠肝脏PAI 1及t PA基因表达改变可能参与NAFLD的发病     

Inhibiting triglyceride synthesis improves hepatic steatosis but exacerbates liver damage and fibrosis in obese mice with nonalcoholic steatohepatitis

In the early stages of nonalcoholic fatty liver disease (NAFLD), triglycerides accumulate in hepatocytes. Diacylglycerol acyltransferase 2 (DGAT2) catalyzes the final step in hepatocyte triglyceride biosynthesis. DGAT2 antisense oligonucleotide (ASO) treatment improved hepatic steatosis dramatically in a previous study of obese mice. According to the 2-hit hypothesis for progression of NAFLD, hepatic steatosis is a risk factor for nonalcoholic steatohepatitis (NASH) and fibrosis. To evaluate this hypothesis, we inhibited DGAT2 in a mouse model of NASH induced by a diet deficient in methionine and choline (MCD). Six-week-old genetically obese and diabetic male db/db mice were fed either the control or the MCD diet for 4 or 8 weeks. The MCD diet group was treated with either 25 mg/kg DGAT2 ASO or saline intraperitoneally twice weekly. Hepatic steatosis, injury, fibrosis, markers of lipid peroxidation/oxidant stress, and systemic insulin sensitivity were evaluated. Hepatic steatosis, necroinflammation, and fibrosis were increased in saline-treated MCD diet-fed mice compared to controls. Treating MCD diet-fed mice with DGAT2 ASO for 4 and 8 weeks decreased hepatic steatosis, but increased hepatic free fatty acids, cytochrome P4502E1, markers of lipid peroxidation/oxidant stress, lobular necroinflammation, and fibrosis. Progression of liver damage occurred despite reduced hepatic expression of tumor necrosis factor alpha, increased serum adiponectin, and striking improvement in systemic insulin sensitivity. CONCLUSION: Results from this mouse model would suggest accumulation of triglycerides may be a protective mechanism to prevent progressive liver damage in NAFLD.     

Expression of p53, Bax and Bcl-2 proteins in hepatocytes in non-alcoholic fatty liver disease

INTRODUCTION Liver steatosis results from triglyceride accumulation in hepatocytes in the course of many diseases[1]. The failure of energetic processes in hepatocytes leads to progressive lipid increase in cytoplasm[2]. The most common cause of liver ste…     

高脂饮食诱导幼鼠脂肪肝模型的建立及其相关代谢研究

目的 通过高脂饮食诱导幼龄大鼠建立非酒精性脂肪性肝病(NAFLD)模型.方法 3周龄刚离乳SD大鼠30只,雌雄各半,随机分为正常(N)组、20%高脂饮食组(HFI组)、30%高脂饮食组(HF2组).无特定病原环境下饲养6周,第6周末处死大鼠,分别进行如下检测:(1)身长、体质量、肝脏质量测量,计算肝指数;(2)采血测空腹ALT、AST、甘油三酯(TG)、总胆固醇(TC)、胰岛素、血糖水平,计算HOMA胰岛素抵抗指数;(3)肝组织匀浆,检测肝脏TG水平;(4)肝组织切片苏木素-伊红染色观察肝组织病理形态,油红O染色显示脂质沉积;(5)免疫组织化学EnVision法染色,显示肝组织固醇调节元件结合蛋白-1、瘦素的表达.组间均数比较采用One-way ANOVA、两两比较采用LSD方法进行统计学分析.结果 病理结果显示模型组大鼠肝脏符合典型的NAFLD病理特征,HF2组内各标本脂肪变性程度较均匀,无动物死亡现象发生;与N组相比,HF2组血清TC显著升高[(2.50±0.39)mmol/L对比(1.82±0.43)mmol/L,P<0.01],肝脏TG含量显著升高[(25.38±13.29)mmol/L对比(12.09±9.59)mmol/L,P<0.01],血清ALT显著升高[(69.80±18.22)U/L对比(48.00±10.45)U/L,P<0.01];与N组相比,HF1组血清TG显著下降[(0.17±0.10)mmol/L对比(0.32±0.12)mmol/L,P<0.05];血糖水平显著升高[(12.33±3.48)mmol/L对比(8.13±2.53)mmol/L,P<0.05];各组血清AST、胰岛素、及HOMA胰岛素抵抗指数差异无统计学意义;模型组肝组织固醇调节元件结合蛋白-1、瘦素表达增加.结论 30%高脂饲料饲养6周,可以诱导幼龄大鼠建立NAFLD模型,高脂饮食导致幼鼠肝脏固醇调节元件结合蛋白-1表达增加,脂肪合成增加,瘦素表达增加.     

Constitutive androstane receptor agonist, TCPOBOP, attenuates steatohepatitis in the methionine choline-deficient diet-fed mouse

AIM： To ascertain whether constitutive androstane receptor （CAR） activation by 1,4-bis-[2-（3,5,- dichloropyridyloxy）] benzene （TCPOBOP） modulates steatohepatitis in the methionine choline-deficient （MCD） diet-fed animal.METHODS： C57/BL6 wild-type mice were fed the MCD or standard diet for 2 wk and were treated with either the CAR agonist, TCPOBOP, or the CAR inverse agonist, androstanol.RESULTS： Expression of CYP2B10 and CYP3A11, known CAR target genes, increased 30-fold and 45-fold, respectively, in TCPOBOP-treated mice fed the MCD diet. TCPOBOP treatment reduced hepatic steatosis （44.6 ＋ 5.4% vs 30.4 ＋ 4.5%, P 〈 0.05） and serum triglyceride levels （48 ＋ 8 vs 20 ＋ 1 mg/dL, P 〈 0.05） in MCD diet- fed mice as compared with the standard diet-fed mice. This reduction in hepatic steatosis was accompanied by an increase in enzymes involved in fatty acid microsomal co-oxidation and peroxisomal p-oxidation, namely CYP4A10, LPBE, and 3-ketoacyI-CoA thiolase. The reduction in steatosis was also accompanied by a reduction in liver cell apoptosis and inflammation. In contrast, androstanol was without effect on any of the above parameters.CONCLUSION： CAR activation stimulates induction of genes involved in fatty acid oxidation, and ameliorates hepatic steatosis, apoptosis and inflammation.     

FXR agonist GW4064 alleviates endotoxin-induced hepatic inflammation by repressing macrophage activation

AIM: To examine the effect of farnesoid X receptor (FXR) activation by GW4064 on endotoxin-induced hepatic inflammation in nonalcoholic fatty liver disease (NAFLD) and the underlying mechanism.METHODS: Six-week-old male C57BL/6 mice were fed a normal diet or a high-fat (HF) diet for 8 wk. HF diet-fed mice were intraperitoneally injected with GW4064 (30 mg/kg) or DMSO (vehicle) once daily for a week and then sacrificed after lipopolysaccharide (LPS, 50 μg/mouse) administration. Hepatic inflammation, levels of the macrophage marker F4/80, and apoptosis were measured at the end of the study. Additionally, the expression of proinflammatory genes involved in NAFLD (interleukin-6, interleukin-1β, interferon-γ, MCP-1) were analyzed by real-time PCR in the murine macrophage cell line RAW 264.7 cultured with or without GW4064 (2 μmol/L) before treatment with LPS.RESULTS: In patients with NAFLD, the expression of FXR was detected by immunohistochemical staining and the relation between FXR expression and NAFLD activity score (NAS) was analyzed. Activation of FXR by GW4064 alleviated hepatic inflammation induced by endotoxin in a murine NAFLD model fed an HF diet as reflected by reduced serum levels of aspartate aminotransferase and alanine aminotransferase. Apoptosis and proinflammatory cytokine levels in liver tissues were also reduced by GW4064, and GW4064 could reduce induction of proinflammatory cytokines by LPS in vitro. FXR levels were reduced in patients with non-alcoholic steatohepatitis compared with healthy controls and were negatively correlated with NAS.CONCLUSION: FXR activation attenuates LPS-induced hepatic inflammation in murine NAFLD by reducing expression of proinflammatory cytokines in macrophages.     

Anti-steatotic and anti-fibrotic effects of the KCa3.1 channel inhibitor,Senicapoc, in non-alcoholic liver disease

AIM To evaluate a calcium activated potassium channel(KCa3.1) inhibitor attenuates liver disease in models of non-alcoholic fatty liver disease(NAFLD).METHODS We have performed a series of in vitro and in vivo studies using the KCa3.1 channel inhibitor, Senicapoc. Efficacy studies of Senicapoc were conducted in toxin-, thioacetamide(TAA) and high fat diet(HFD)-induced models of liver fibrosis in rats. Efficacy and pharmacodynamic effects of Senicapoc was determined through biomarkers of apoptosis, inflammation, steatosis and fibrosis. RESULTS Upregulation of KCa3.1 expression was recorded in TAA-induced and high fat diet-induced liver disease. Treatment with Senicapoc decreased palmitic aciddriven Hep G2 cell death.(P 0.05 vs control) supporting the finding that Senicapoc reduces lipiddriven apoptosis in Hep G2 cell cultures. In animals fed a HFD for 6 wk, co-treatment with Senicapoc,(1) reduced non-alcoholic fatty liver disease(NAFLD) activity score(NAS)(0-8 scale),(2) decreased steatosis and(3) decreased hepatic lipid content(Oil Red O, P 0.05 vs vehicle). Randomization of TAA animals and HFD fed animals to Senicapoc was associated with a decrease in liver fibrosis as evidenced by hydroxyproline and Masson's trichrome staining(P 0.05 vs vehicle). These results demonstrated that Senicapoc mitigates both steatosis and fibrosis in liver fibrosis models.CONCLUSION These data suggest that Senicapoc interrupts more than one node in progressive fatty liver disease by its anti-steatotic and anti-fibrotic activities, serving as a double-edged therapeutic sword.     

Genetic differences in oxidative stress and inflammatory responses to diet‐induced obesity do not alter liver fibrosis in mice

Objective: To determine how genetic factors might influence the progression of nonalcoholic fatty liver disease (NAFLD). Design/Intervention: Beginning in adolescence, male C57BL6 (BL6) and 129/SVJ mice were fed control (n=15/group) or high‐fat (HF) diets (n=30/group) for 6 months. Main Outcome Measures: Assessed were body weight, insulin resistance, hepatic production of free radicals, expression of cytokines and fibrosis‐related genes and severity of hepatic steatosis, injury and fibrosis. Results: High‐fat diets induced comparable obesity, hepatic steatosis and insulin resistance in the two strains. Compared with BL6 mice, 129/SVJ mice had impaired induction of antioxidant genes, generated three‐ to four‐fold more free radicals and exhibited two‐fold greater induction of profibrogenic cytokines (interleukin‐4 and transforming growth factor‐β1) and fibrosis‐related genes (fibronectin and tissue inhibitor of metalloproteinase‐1) (all P<0.05 for 129 vs BL6). Surprisingly, however, induction of collagen I α1 mRNA and accumulation of Sirius red‐stained fibrils and hepatic hydroxyproline were similar in BL6 and 129/SVJ mice, and although patchy sinusoidal fibrosis emerged in both strains, neither developed bridging fibrosis. Conclusions: Although BL6 and 129/SVJ mice with diet‐induced obesity, insulin resistance and steatosis differed with respect to several factors that are thought to influence human NAFLD progression, they developed comparable liver fibrosis. Moreover, none of the risk factors for NAFLD‐related cirrhosis in humans, including obesity, insulin resistance, chronic inflammatory and oxidant stress, steatohepatitis or activation of fibrogenic genes, proved to be sufficient to cause cirrhosis in these mice, even when exposure to one or more of these insults was very prolonged.     

Role of EpCAM+ CD133+ extracellular vesicles in steatosis to steatohepatitis transition in NAFLD

Background and Aims

Extracellular vesicles (EVs) have emerged as a potential source of circulating biomarkers in liver disease. We evaluated circulating AV+ EpCAM+ CD133+ EVs as a potential biomarker of the transition from simple steatosis to steatohepatitis.

Methods

EpCAM and CD133 liver proteins and EpCAM+ CD133+ EVs levels were analysed in 31 C57BL/6J mice fed with a chow or high fat, high cholesterol and carbohydrates diet (HFHCC) for 52 weeks. The hepatic origin of MVs was addressed using AlbCrexmT/mG mice fed a Western (WD) or Dual diet for 23 weeks. Besides, we assessed plasma MVs in 130 biopsy-proven NAFLD patients.

Results

Hepatic expression of EpCAM and CD133 and EpCAM+ CD133+ EVs increased during disease progression in HFHCC mice. GFP+ MVs were higher in AlbCrexmT/mG mice fed a WD (5.2% vs 12.1%) or a Dual diet (0.5% vs 7.3%). Most GFP+ MVs were also positive for EpCAM and CD133 (98.3% and 92.9% respectively), suggesting their hepatic origin. In 71 biopsy-proven NAFLD patients, EpCAM+ CD133+ EVs were significantly higher in those with steatohepatitis compare to those with simple steatosis (286.4 ± 61.9 vs 758.4 ± 82.3; p < 0.001). Patients with ballooning 367 ± 40.6 vs 532.0 ± 45.1; p = 0.01 and lobular inflammation (321.1 ± 74.1 vs 721.4 ± 80.1; p = 0.001), showed higher levels of these EVs. These findings were replicated in an independent cohort.

Conclusions

Circulating levels of EpCAM+ CD133+ MVs in clinical and experimental NAFLD were increased in the presence of steatohepatitis, showing high potential as a non-invasive biomarker for the evaluation and management of these patients.