DOI:10.20047/j.issn1673-7210.25080587
中图分类号:R259
王志文, 丘清元, 吴雅静, 叶玉静, 林渝, 蒋筱
| 【作者机构】 | 广西中医药大学 |
| 【分 类 号】 | R259 |
| 【基 金】 | 国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023167) 广西中医药大学赛恩斯新医药学院自然科学研究项目(2024ZZB009) 广西中医药大学赛恩斯新医药学院校级科研项目(2022MS009)。 |
肝纤维化是慢性肝病进展中的关键病理过程,以细胞外基质过度沉积为特征,可导致肝硬化、肝衰竭,甚至肝癌,严重危害患者健康。早期精准诊断对延缓疾病进展、改善预后至关重要。肝活检作为传统金标准,因侵入性操作风险及取样误差等问题,临床应用受限。随着影像技术、分子生物学及人工智能的迅速发展,无创诊断技术成为研究热点。多模态影像组学、液体活检、人工智能决策系统等新兴方法为肝纤维化的早期、精准与动态评估提供新途径。
肝纤维化是肝脏对慢性损伤的一种修复性反应,其病理过程涉及多种细胞类型介导的复杂信号通路[1-2]。在生理状态下,肝星状细胞维持静息状态;当肝脏受损时,肝星状细胞则被激活并转化为肌成纤维细胞,激活的肝星状细胞发生显著增殖并大量合成与分泌细胞外基质成分,导致细胞外基质过度沉积,进而破坏肝脏的正常微结构和功能[3-5]。
在肝纤维化的分子机制中,肝星状细胞的激活是核心事件。极光激酶A可通过激活Wnt/β-catenin信号通路驱动肝星状细胞活化,并促进肝纤维化进展。靶向抑制极光激酶A活性可减轻肝纤维化程度[6]。过表达甲基胞嘧啶双加氧酶3可诱导肝星状细胞铁死亡,通过调控铁死亡相关基因甲基化以缓解纤维化[7]。研究显示,肝损伤后肝细胞发生代谢重编程,以满足活化成纤维细胞的能量需求,提示能量代谢在纤维化发展中起重要作用[8]。
长链非编码RNA(longnoncodingRNA,lncRNA)-Gm 9866在肝纤维化中发挥重要作用。lncRNA-Gm9866在活化的肝星状细胞及纤维化小鼠肝脏模型中的表达显著上调,其过表达可促进肝星状细胞活化和肝细胞凋亡;反之,沉默该基因则抑制肝星状细胞的活化及转化生长因子(transforming growth factor,TGF)-β1诱导的纤维化。机制上,lncRNA-Gm9866可能通过靶向Fam98b蛋白,进而调控TGF-β/Smad、Notch信号通路。因此,lncRNA-Gm9866/Fam98b轴可作为肝纤维化诊疗的新潜在靶点[9]。
无创诊断技术的发展可显著改变肝纤维化的诊断方式。传统上,肝纤维化诊断主要依赖肝活检,虽能提供准确病理信息,但其侵入性操作伴随创伤、并发症风险、取样误差和患者接受度低等局限[10]。近年来,基于血清学指标的无创方法通过检测特定生物标志物用于临床评估,但单一指标常因敏感度与特异度不足难以准确反映纤维化程度[11]。影像学技术如瞬时弹性成像通过测量肝组织硬度实现快速、简便的诊断,对肝硬化的敏感度和特异度分别达87%和91%[12-13]。相比之下,磁共振弹性成像能更精确评估纤维化,尤其在早期阶段优势明显[12]。除血清学与影像学技术外,基于分子生物学方法,检测循环微RNA(microRNA,miRNA)的表达变化成为研究热点,显示出作为无创生物标志物的潜力,这些进展共同推动肝纤维化诊断从侵入性向无创性的根本转变[14-15]。
影像组学通过高通量提取并分析医学图像中的定量特征,结合机器学习算法识别与肝纤维化相关的影像标志物,为疾病诊断与分期提供客观、量化的依据[16]。基于超声、计算机体层成像(computed tomography,CT)和磁共振成像(magnetic resonance imaging,MRI)的影像组学研究在肝纤维化评估中取得初步成效。一项基于232例患者多期相CT影像特征的研究采用CatBoost算法构建机器学习模型,用于预测肝纤维化分期,显示该模型在不同分期中的曲线下面积为0.65~0.80,性能优于纤维化-4指数的方法,且在区分晚期纤维化和肝硬化方面优于放射科医师视觉评估[17]。在MRI影像组学领域,一项MRI影像组学研究通过对非增强T1 加权和T2 加权脂肪饱和图像进行纹理分析并结合机器学习,在区分早、晚期肝纤维化方面准确率达85.7%,诊断性与磁共振弹性成像相当[18]。
多模态影像技术通过整合不同影像模态的互补优势,以提高肝纤维化诊断的准确性和全面性。常见组合包括正电子发射计算机断层成像(positron emission tomography,PET)/CT、MRI与超声等[19]。PET/CT结合PET的功能代谢信息与CT的解剖结构信息,除广泛应用于肿瘤诊断与分期外,在肝纤维化评估中展现出潜在价值。MRI与超声联合则融合前者优异的软组织分辨能力和后者实时、便捷与经济性。研究通过结合MRI扩散加权成像与超声弹性成像,分别从水分子扩散受限和肝脏硬度两个维度评估纤维化,显著提升诊断效能。然而,多模态融合仍面临图像配准与数据融合等技术挑战,需确保不同模态图像的空间对齐与信息有效整合。
液体活检标志物在肝纤维化诊断与监测中具有重要潜力[20]。血清标志物如透明质酸、Ⅲ型前胶原氨基端肽和基质金属蛋白酶等表现出良好的诊断性能[21-22]。纤维化-4指数、天冬氨酸氨基转移酶/血小板比值指数在预测慢性肝病患者显著纤维化和肝硬化方面表现出较高的诊断准确性[23-24]。
透明质酸水平与纤维化程度呈正相关,能有效区分不同程度的纤维化[25]。结合层粘连蛋白可进一步提高准确性,其修饰的药物递送系统显示出治疗潜力[26-27]。血清Ⅲ型前胶原氨基端肽是多变量分析中独立的预测因子,对小儿胆汁淤积及非酒精性脂肪性肝病相关肝纤维化具有突出诊断价值,其在区分肝纤维化分期时表现出较高的受试者操作特征曲线下面积及优异的敏感度和特异度[28]。在非酒精性脂肪性肝病儿童中,与血小板比值指数和纤维化-4指数比较,Ⅲ型前胶原氨基端肽能更准确地评估纤维化严重程度,其水平升高与F≥2纤维化风险显著相关,展现出良好的非侵入性生物标志物潜力[29]。基质金属蛋白酶在肝纤维化过程中的表达和作用呈阶段性变化。早期某些基质金属蛋白酶表达上调可激活肝星状细胞,并促进纤维化进展;后期基质金属蛋白酶下调导致细胞外基质积累和疾病恶化[30]。除降解细胞外基质外,基质金属蛋白酶还参与调控细胞增殖、基因表达及凋亡等多种生物学过程[31]。血清基质金属蛋白酶-7与非酒精性脂肪性肝病患者显著纤维化独立相关,有助于提升诊断效能,显示出其作为生物标志物和治疗靶点的潜在价值[32]。
一项多中心研究基于蛋白质组学和流式细胞术检测250例经组织学证实的非酒精性脂肪性肝病/非酒精性脂肪性肝炎患者单核细胞中PLIN2与RAB14表达水平,显示PLIN2平均荧光强度联合腰围、甘油三酯、丙氨酸氨基转移酶及糖尿病状态诊断非酒精性脂肪性肝炎的准确率在发现和验证队列中分别达93%和92%;RAB14平均荧光强度联合年龄、腰围、高密度脂蛋白胆固醇、血糖及丙氨酸氨基转移酶诊断肝纤维化的曲线下面积分别为95.9%和99.3%,该标志物的诊断效能显著优于常用无创指标[33]。
血清中特定miRNA如miR-138、miR-140、miR-143等在慢性丙型肝炎相关肝纤维化患者中的表达上调。早期(F0~F2)和晚期(F3~F4)纤维化患者高于健康对照组,miR-138在早、晚期纤维化中敏感度均为89.3%,特异度分别为71.4%和93.0%;miR-143在晚期纤维化中敏感度和特异度分别为75.0%和88.4%,这些miRNA显示出作为肝纤维化诊断与监测标志物的潜力[15]。
在肝纤维化诊断领域,液体活检技术有望在多个方面取得重要突破。①技术创新:新兴检测技术可显著提升液体活检的准确度与敏感度,如纳米技术为循环肿瘤细胞及胞外囊泡的检测与分子表征提供新策略。纳米材料与纳米结构的进展可增强循环肿瘤细胞/胞外囊泡检测的敏感度、特异度及纯度;新型传感与可视化技术则助力其精准分析[34]。②新型标志物探索:除现有标志物外,细胞游离DNA甲基化模式的深入研究有望开发高特异度诊断及预后标志物。细胞游离DNA甲基化状态与肿瘤进展密切相关,其在肝纤维化中可能具有重要价值,通过分析细胞游离DNA甲基化谱有助于实现早期诊断和病情监测[35-36]。③多技术整合:液体活检与多模态影像组学及人工智能相结合,可实现从分子到影像的多维度评估。人工智能算法有助于挖掘生物标志物与疾病的关联,提升诊断准确性。未来仍需推进检测标准化并进一步提高标志物的特异度和敏感度,以促进临床转化[37]。
人工智能技术在肝纤维化无创诊断领域进展显著。通过分析临床数据挖掘潜在规律,为诊断与评估提供客观依据[26]。在影像分析方面,人工智能可结合超声、CT和MRI等技术,自动识别和量化纤维化程度,从而减少对侵入性活检的依赖[38]。基于人工智能的二次谐波生成/双光子激发荧光显微镜技术能通过机器学习算法精准量化肝胶原形态,提供连续纤维化指数,克服传统病理的变异性[39]。一项研究开发基于钆塞酸增强肝胆期MRI的全自动深度学习算法,与磁共振弹性成像比较,用于无创肝纤维化分期,该研究纳入355例患者显示,深度学习算法在训练集、验证集与测试集中对纤维化分期(F1~F4、F2~F4、F3~F4、F4)的预测曲线下面积分别为0.99/0.70/0.77、0.92/0.71/0.91、0.91/0.78/0.90、0.98/0.83/0.85,其诊断性能与磁共振弹性成像相当,提示基于钆塞酸增强肝胆期MRI的全自动深度学习模型在肝纤维化分期中具有良好至优秀的诊断效能[40]。
基于临床参数的人工智能整合同样具有价值。一项涵盖19项研究的荟萃分析显示,人工智能模型诊断肝硬化的合并敏感度和特异度分别为0.78和0.89,诊断晚期纤维化(≥F3)分别为0.86和0.87,诊断显著纤维化(≥F2)分别为0.86和0.81,且其他指标如阳性预测值、阴性预测值及诊断比值均表现稳健,该结果证实其具有可靠的诊断潜力[41]。
肝纤维化无创诊断技术的临床转化面临多重挑战。技术上,血液中循环肿瘤细胞和外泌体等生物标志物水平极低,现有检测方法在敏感度、特异度及可重复性方面存在不足,且缺乏标准化流程,导致结果难以比较和解读[42]。临床验证则受限于大规模试验招募难、疾病异质性及混杂变量多,影响技术的普适性和准确性[43]。部分高新技术如纳米检测或高端影像设备成本高昂,制约其临床普及与经济可行性[44]。
在肝纤维化无创诊断中,不同方法的准确性与可靠性尚存争议。肝活检作为传统金标准,因侵入性、取样误差及操作风险使其应用受限[10]。瞬时弹性成像和血清标志物等无创方法虽更安全便捷,但准确性仍待提高。瞬时弹性成像在病毒性肝炎相关肝硬化中诊断性能优于血清学指标,两者联用可减少对肝活检的需求[45]。但在肥胖患者中可能无法获得可靠的肝硬度测量,在非酒精性脂肪性肝病中受脂肪变性影响[46]。联合肝硬度测量与非酒精性脂肪性肝病纤维化评分可提升诊断准确性,但仍需依据群体特点个性化应用[47]。目前建议联合多项无创技术进行初筛,结果不确定时再考虑肝活检,以兼顾安全性与诊断精确度。
肝纤维化无创诊断技术持续创新,为早期精准识别提供多种新途径。分子MRI通过靶向特定分子探针可视化纤维化相关靶点,可在动物模型中检测早期微小病变,从分子机制层面提升诊断特异度和敏感度[48]。超声技术中,二维剪切波弹性成像可准确地测量肝硬度,其诊断显著纤维化(≥F2)和肝硬化(F4)的曲线下面积分别达0.862和0.926,性能优于传统方法[49]。在血清生物标志物方面,平滑肌肌动蛋白α 和卵泡抑素样蛋白-1在早期诊断中显示出较高敏感度与特异度[45,50]。此外,基于纳米技术的生物传感器能高灵敏检测体液中的miRNA、蛋白质等标志物,有助于实现早期诊断与动态监测。尽管这些无创方法取得显著进展,未来仍需优化各技术在不同病程阶段的准确性,通过多手段联合应用进一步提升诊断效能,减少对肝活检的依赖[51]。
利益冲突声明:本文所有作者均声明不存在利益冲突。
[1] SHU Y,HE Y,YE G,et al.Curcumin inhibits the activity and induces apoptosis of activated hepatic stellate cell by suppressing autophagy [J].J Cell Biochem,2023,124(11):1764-1778.
[2] YUAN M,YAO L,CHEN P,et al.Human umbilical cord mesenchymal stem cells inhibit liver fibrosis via the micro RNA-148a-5p/SLIT3 axis[J].Int Immunopharmacol,2023,125:111134.
[3] DONG J,VISWANATHAN S,ADAMI E,et al.Hepatocytespecific IL11 cis-signaling drives lipotoxicity and underlies the transition from NAFLD to NASH[J].Nat Commun,2021,12(1):66.
[4] CLAVERIA-CABELLO A,COLYN L,URIARTE L,et al.Dual pharmacological targeting of HDACs and PDE5 inhibits liver disease progression in a mouse model of biliary inflammation and fibrosis [J].Cancers(Basel),2020,12(12):3748.
[5] ZHANG L,ZHOU Q,ZHANG J,et al.Liver transcriptomic and proteomic analyses provide new insight into the pathogenesis of liver fibrosis in mice[J].Genomics,2023,115(6):110738.
[6] DAI G,LIN J,JIANG Y,et al.Aurora kinase A promotes hepatic stellate cell activation and liver fibrosis through the Wnt/β-catenin pathway[J].Front Oncol,2025,14:1517226.
[7] LIU Y,FENG L L,HAN B,et al.Exploring the molecular mechanisms through which overexpression of TET3 alleviates liver fibrosis in mice via ferroptosis in hepatic stellate cells[J].Cell Signal,2025,131:111747.
[8] IRSHAD I,ALQAHTANI S A,IKEJIMA K,et al.Energy metabolism:an emerging therapeutic frontier in liver fibrosis [J].Ann Hepatol,2025,30(1):101896.
[9] LIAO X,RUAN X,YAO P,et al.LncRNA-Gm9866 promotes liver fibrosis by activating TGFβ/Smad signaling via targeting Fam98b[J].J Transl Med,2023,21(1):778.
[10] CHOU Y T,LI C H,SUN Z J,et al.A positive relationship between betel nut chewing and significant liver fibrosis in NAFLD subjects,but not in non-NAFLD ones [J].Nutrients,2021,13(3):914.
[11] XU B,ZHOU N M,CAO W T,et al.Evaluation of elastography combined with serological indexes for hepatic fibrosis in patients with chronic hepatitis B [J].World J Gastroenterol,2018,24(37):4272.
[12] ZERUNIAN M,PUCCIARELLI F,MASCI B,et al.Updates on quantitative MRI of diffuse liver disease:a narrative review[J].Biomed Res Int,2022,2022:1147111.
[13] CHENG H S,RADEMAKER M.Monitoring methotrexateinduced liver fibrosis in patients with psoriasis:utility of transient elastography[J].Psoriasis(Auckl),2018,8:21-29.
[14] FANG Z,DOU G,WANG L.MicroRNAs in the pathogenesis of nonalcoholic fatty liver disease[J].J Clin Med,2021,17(7):1851.
[15] EL-AHWANY E,NAGY F,ZOHEIRY M,et al.Circulating miRNAs as predictor markers for activation of hepatic stellatecellsandprogressionofHCV-inducedliverfibrosis[J].Electron Physician,2016,8(1):1804.
[16] SUNG Y S,PARK B,PARK H J,et al.Radiomics and deep learning in liver diseases[J].J Gastroenterol Hepatol,2021,36(3):561-568.
[17] CUI E,LONG W,WU J,et al.Predicting the stages of liver fibrosis with multiphase CT radiomics based on volumetric features[J].Abdom Radiol(NY),2021,46(8):3866-3876.
[18] SCHAWKAT K,CIRITSIS A,VON ULMENSTEIN S,et al.Diagnostic accuracy of texture analysis and machine learning for quantification of liver fibrosis in MRI:correlation with MR elastography and histopathology [J].Eur Radiol,2020,30(8):4675-4685.
[19] TAE C H,LEE J H,CHOI J Y,et al.Impact of incidental findings on integrated 2-[18F]-fluoro-2-deoxy-D-glucose positron emission tomography/computed tomography in patients with gastric cancer[J].Asia Pac J Clin Oncol,2015,11(1):34-40.
[20] WANGC,ZHANG D,YANG H,et al.A light-activated magnetic bead strategy utilized in spatio-temporal controllable exosomes isolation [J].Front Bioeng Biotechnol,2022,10:1006374.
[21] IRVINE K M,WOCKNER L F,HOFFMANN I,et al.Multiplex serum protein analysis identifies novel biomarkers of advanced fibrosis in patients with chronic liver disease with the potential to improve diagnostic accuracy of established biomarkers[J].PLoS One,2016,11(11):e0167001.
[22] CHROSTEK L,PANASIUK A.Liver fibrosis markers in alcoholic liver disease [J].World J Gastroenterol,2014,20(25):8018-8023.
[23] LIN C L,LIU C H,WANG C C,et al.Serum biomarkers predictive of significant fibrosis and cirrhosis in chronic hepatitis B[J].J Clin Gastroenterol,2015,49(8):705-713.
[24] ZENG S,LIU Z,KE B,et al.The non-invasive serum biomarkers contributes to indicate liver fibrosis staging and evaluate the progress of chronic hepatitis B[J].BMC Infect Dis,2024,24(1):638.
[25] LI F,ZHU C L,ZHANG H,et al.Role of hyaluronic acid and laminin as serum markers for predicting significant fibrosis in patients with chronic hepatitis B [J].Braz J Infect Dis,2012,16(1):9-14.
[26] WANG C,CUI J J,SHENG T,et al.Chitosan-decorated silibinin-hyaluronic acid conjugate for enhancing intestinal absorption and improving liver fibrosis[J].Int J Biol Macromol,2025,320(Pt 4):146057.
[27] YU F,LIU Z,FENG J,et al.Hyaluronic acid modified extracellular vesicles targeting hepatic stellate cells to attenuate hepatic fibrosis[J].Eur J Pharm Sci,2024,198:106783.
[28] WANG Y,PAN W,ZHAO D,et al.Diagnostic value of serum procollagen ⅢN-terminal peptide for liver fibrosis in infantile cholestasis[J].Front Pediatr,2020,8:131.
[29] MOSCA A,COMPARCOLA D,ROMITO I,et al.Plasma Nterminal propeptide of type Ⅲprocollagen accurately predicts liver fibrosis severity in children with non-alcoholic fatty liver disease[J].Liver Int,2019,39(12):2317-2329.
[30] SABIRU,GUH,ZHANGDW.Extracellularmatrixturnover:phytochemicals target and modulate the dual role of matrix metalloproteinases(MMPs)in liver fibrosis [J].Phytother Res,2023,37(11):4932-4962.
[31] RODERFELD M.Matrix metalloproteinase functions in hepatic injury and fibrosis[J].Matrix Biol,2018,68:452-462.
[32] IRVINE K M,OKANO S,PATEL P J,et al.Serum matrix metalloproteinase 7(MMP7)is a biomarker of fibrosis in patients with non-alcoholic fatty liver disease[J].Sci Rep,2021,11(1):2858.
[33] ANGELINIG,PANUNZIS,CASTAGNETO-GISSEYL,et al.Accurate liquid biopsy for the diagnosis of non-alcoholic steatohepatitis and liver fibrosis[J].Gut,2023,72(2):392-403.
[34] LI W,WANG H,ZHAO Z,et al.Emerging nanotechnologies for liquid biopsy:the detection of circulating tumor cells and extracellular vesicles [J].Adv Mater,2019,31(45):1805344.
[35] ZHOU X,CHENG Z,DONG M,et al.Tumor fractions deciphered from circulating cell-free DNA methylation for cancer early diagnosis[J].Nat Commun,2022,13(1):7694.
[36] GAI W,SUN K.Epigenetic biomarkers in cell-free DNA and applications in liquid biopsy [J].Genes(Basel),2019,10(1):32.
[37] POWROZEK T,OCHIENG OTIENO M.Blood circulating non-coding RNAs for the clinical management of triplenegative breast cancer [J].Cancers(Basel),2022,14(3):803.
[38] YIN C,ZHANG H,DU J,et al.Artificial intelligence in imaging for liver disease diagnosis [J].Front Med(Lausanne),2025,12:1591523.
[39] AKBARY K,NOUREDDIN M,YAYUN R,et al.Development of AI based fibrosis detection algorithm by SHG/TPEF microscopy for fully quantified liver fibrosis assessment in MASH[J].Liver Int,2025,45(9):e70258.
[40] HECTORS S J,KENNEDY P,HUANG K H,et al.Fully automated prediction of liver fibrosis using deep learning analysis of gadoxetic acid-enhanced MRI [J].Eur Radiol,2021,31(6):3805-3814.
[41] DECHARATANACHARTP,CHAITEERAKIJR,TIYARATTANACHAI T,et al.Application of artificial intelligence in chronic liver diseases:a systematic review and metaanalysis[J].BMC Gastroenterol,2021,21(1):10.
[42] SHAO H,CHUNG J,ISSADORE D.Diagnostic technologies for circulating tumour cells and exosomes [J].Biosci Rep,2016,36(1):e00292.
[43] AMADI C N,ORISAKWE O E.Herb-induced liver injuries in developing nations:an update[J].Toxics,2018,6(2):24.
[44] COMBES G F,VU
KOVIC' A M,PERIC' BAKULIC' M,et al.Nanotechnology in tumor biomarker detection:the potential of liganded nanoclusters as nonlinear optical contrast agents for molecular diagnostics of cancer [J].Cancers(Basel),2021,13(16):4206.
[45] LI W,CHI Y,XIAO X,et al.Plasma FSTL-1 as a noninvasive diagnostic biomarker for patients with advanced liver fibrosis[J].Hepatology,2025,82(3):669-682.
[46] CASTERA L.Noninvasive assessment of liver fibrosis[J].Dig Dis,2015,33(4):498-503.
[47] JOO S K,KIM W,KIM D,et al.Steatosis severity affects the diagnostic performances of noninvasive fibrosis tests in nonalcoholic fatty liver disease [J].Liver Int,2018,38(2):331-341.
[48] LI Z,SUN J,YANG X.Recent advances in molecular magnetic resonance imaging of liver fibrosis[J].Biomed Res Int,2015,2015:595467.
[49] ZHENG J,GUO H,ZENG J,et al.Two-dimensional shearwave elastography and conventional US:the optimal evaluation of liver fibrosis and cirrhosis[J].Radiology,2015,275(1):290-300.
[50] CARDOSO-LEZAMAI,RAMOS-TOVARE,ARELLANESROBLEDO J,et al.Serum α-SMA is a potential noninvasive biomarker of liver fibrosis [J].Toxicol Mech Methods,2024,34(1):13-19.
[51] LOOMBAR,ADAMS LA.Advancesin non-invasiveassessment of hepatic fibrosis[J].Gut,2020,69(7):1343-1352.
Research progress on non-invasive and accurate diagnostic technology of hepatic fibrosis
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