Ferroptosis is an iron‑dependent form of cell death associated with liver pathologies. However, its role in chronic cholestasis remains to be fully elucidated. The present study therefore investigated the pathological mechanism of ferroptosis in a rat model of α‑naphthyl isothiocyanate (ANIT)‑induced chronic cholestasis and evaluated the therapeutic potential of the iron chelator deferoxamine (DFO). Wistar rats were used to establish a chronic cholestasis model via ANIT administration, with a subset of animals receiving DFO treatment. Wistar rats that were subjected to chronic ANIT exposure were found to develop severe liver injury, characterized by impaired function, inflammation and fibrosis. In addition, pronounced iron deposition and hallmark features of ferroptosis, including elevated lipid peroxidation, depleted glutathione, and aberrant expression of acyl‑CoA synthetase long‑chain family member 4 and cyclooxygenase 2, were observed. Ultrastructural analysis revealed distinctive mitochondrial abnormalities consistent with ferroptosis. Mechanistically, these changes appeared to be mediated by suppression of the Kelch‑like ECH‑associated protein 1/nuclear factor erythroid 2‑related factor 2/heme oxygenase 1 antioxidant pathway and dysregulation of key iron metabolism proteins, including transferrin receptor 1 and ferroportin 1. Intervention with DFO markedly ameliorated the cholestatic injury, reduced iron overload and lipid peroxidation, mitigated mitochondrial damage, and normalized the expression of key proteins involved in ferroptosis, antioxidant defense and iron homeostasis. Taken together, these findings suggested that ferroptosis may be a key pathological mechanism in chronic cholestasis, driven by the concurrent disruption of antioxidant and iron metabolic capacities in hepatocytes. Therefore, targeting iron overload may be a promising therapeutic strategy for cholestasis. .

Accurate, non-invasive liver fibrosis detection is essential for chronic liver disease management, particularly with rising metabolic dysfunction-associated liver disease (MASLD) and chronic hepatitis B (CHB). While the Fibrosis-4 (FIB-4) index is widely used, its performance for advanced fibrosis is limited. We develop Met-FIB using metabolomics and machine learning, integrating FIB-4 parameters (age, aspartate aminotransferase, alanine aminotransferase, and platelet count) with tyrosine and taurocholic acid identified in a CHB discovery cohort (n = 3,251). Validation includes one CHB cohort (n = 729) and two MASLD cohorts (n = 149, n = 155). Met-FIB outperforms FIB-4, FibroScan, and other serum markers across all fibrosis stages. In CHB, Met-FIB achieves 96.3% rule-out sensitivity and 85.4% rule-in specificity for significant fibrosis, with rule-in specificity reaching 98.6% and 98.8% for advanced fibrosis and cirrhosis. In MASLD, corresponding values are 93.9% and 90.2% for significant fibrosis, with >97.9% specificity for late-stage disease. Met-FIB demonstrates clinical utility for non-invasive fibrosis staging across diverse etiologies.

The conventional biomarker discovery approach in metabolomics treats metabolites as isolated features, failing to capture their interdependencies. We applied a network-based method using Tabular Prior-Data Fitted Networks (TabPFN) integrated with SHapley Additive exPlanations (SHAP) to derive stage-specific metabolic networks capturing statistical metabolite co-contribution patterns and identify key metabolites. We analyzed 1312 participants across five stages (control, hepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma (HCC)), quantitatively profiling 160 metabolites, and externally validated the results in 721 samples. We compared three approaches: (1) TabPFN-SHAP, (2) conventional statistics with machine learning, and (3) TabPFN-SHAP integrated with literature-validated biomarkers. TabPFN-SHAP–based approaches outperformed the conventional pipeline, revealing distinct stage-specific metabolic signatures with overlapping metabolites between consecutive networks, marking consistently prioritized features. A five-metabolite HCC-screening panel (sebacic acid, alpha-linolenic acid, phenylalanyl-tryptophan, arachidonic acid, and glycocholic acid) was identified. At the screening-oriented cutoff, the panel achieved an AUROC of 0.979 (95% CI: 0.966–0.991) for Control vs HCC and 0.955 (95% CI: 0.936–0.977) for Fibrosis/Cirrhosis vs HCC, with sensitivity of 0.937, specificity ≥0.855, accuracy ≥0.897, and negative predictive value ≥ 0.981 across both clinical scenarios, outperforming all comparative approaches. TabPFN-SHAP provides an interpretable computational framework for metabolite-panel discovery, with demonstrated utility for HCC screening.

The global incidence of metabolic dysfunction-associated fatty liver disease (MAFLD) is increasing annually, which has become a major public-health concern. MAFLD is typically associated with obesity, hyperlipemia, or metabolic syndrome. Dietary induction is one of the most common methods for preparing animal models of MAFLD. However, there are phenotypic differences between methionine-choline-deficient diet (MCDD) and high fat diet (HFD) models. To explore the differences in hepatic fatty acid metabolism between MCDD and HFD induced MAFLD, we analyzed serum and liver tissue from the two MAFLD models. We found that liver fat accumulation and liver function damage were common pathological features in both MAFLD models. Furthermore, in the MCDD model, the expression of hepatic fatty acid transport proteins increased, while the expression of hepatic fatty acid efflux proteins and mRNA decreased, along with a decrease in blood lipid levels. In the HFD model, the expression of hepatic fatty acid uptake proteins, efflux proteins and efflux mRNA increased, along with an increase in blood lipid levels. Impaired fatty acid oxidation and increased hepatic fatty acid uptake play key roles in the pathogenesis of the two MAFLD models. The inverse changes in de novo lipogenesis and fatty acid efflux may represent an important pathological mechanism that leads to the phenotypic differences between the MCDD and HFD models.

Metabolic dysfunction-associated steatotic liver disease (MASLD), as the hepatic manifestation of metabolic syndrome, is characterized by hepatic steatosis, inflammation, and fibrosis. Accumulating evidence suggests that metabolic reprogramming, particularly enhanced glycolysis, plays an important role in the pathogenesis of MASLD. Dysregulated glycolysis not only supplies energy and biosynthetic precursors for multiple hepatic cell types but also directly contributes to lipid accumulation, inflammatory activation, fibrogenesis, and even hepatocellular carcinogenesis. This review summarizes the mechanisms underlying glycolytic reprogramming in MASLD and examines the distinct roles of glycolytic alterations in various cell types, including hepatocytes, macrophages, hepatic stellate cells (HSCs), T cells, and bile duct epithelial cells (BECs). Furthermore, glycolysis mediates the crosstalk between different cell types and organ types in MASLD, modulating cytokine production and activating pro-inflammatory and pro-fibrotic signaling pathways. Finally, this article discusses the potential and challenges of targeting key glycolytic enzymes - such as hexokinase 2 (HK2), 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase (PFKFB3) and Pyruvate kinase M2 (PKM2) - as therapeutic strategies, offering new perspectives for alleviating the disease burden of MASLD.

Background: The aim of this study was to evaluate the accuracy of LiveBoost™, a gradient boosting (GB)-based prediction system based on standard biochemical values (AST, ALT, platelet count) and age, in Chinese patients with chronic hepatitis B (CHB) and compare its performance with FIB-4 (fibrosis-4 score) and APRI (the aspartate transaminase to platelet ratio index). Methods: This retrospective trial enrolled 454 participants, including 279 CHB patients who underwent liver biopsy and 175 normal controls from 3 centers in China. All participants underwent laboratory blood testing. LiveBoost was constructed using GB and FIB-4 and APRI were calculated from laboratory data. Results: LiveBoost outperformed APRI and FIB-4 in predicting hepatic fibrosis and cirrhosis. The GB model had an AUROC of 0.977 for CHB diagnosis, 0.804 for early and advanced fibrosis, and 0.836 for non-cirrhosis and cirrhosis, compared to AUROC of 0.554, 0.673 and 0.720 for FIB-4, AUROC of 0.977, 0.652 and 0.654 for APRI. Conclusions: LiveBoost is a more reliable and cost-effective method than APRI and FIB-4 for assessing liver fibrosis in Chinese patients with CHB.

Consumption of herbal products containing pyrrolizidine alkaloids (PAs) is one of the major causes for hepatic sinusoidal obstruction syndrome (HSOS), a deadly liver disease. However, the crucial metabolic variation and biomarkers which can reflect these changes remain amphibious and thus to result in a lack of effective prevention, diagnosis and treatments against this disease. The aim of the study was to determine the impact of HSOS caused by PA exposure, and to translate metabolomics-derived biomarkers to the mechanism. In present study, cholic acid species (namely, cholic acid, taurine conjugated-cholic acid, and glycine conjugated-cholic acid) were identified as the candidate biomarkers (area under the ROC curve 0.968 [95

Gardenia jasminoides Ellis, a staple in herbal medicine, has long been esteemed for its purported hepatoprotective properties. Its primary bioactive constituent, geniposide, has attracted considerable scientific interest owing to its multifaceted therapeutic benefits across various health conditions. However, recent investigations have unveiled potential adverse effects associated with its metabolite, genipin, particularly at higher doses and prolonged durations of administration, leading to hepatic injury. Determining the optimal dosage and duration of geniposide administration while elucidating its pharmacological and toxicological mechanisms is imperative for safe and effective clinical application.This study aimed to evaluate the safe dosage and administration duration of geniposide in mice and investigate its toxicological mechanisms within a comprehensive dosage-duration-efficacy/toxicity model. Four distinct mouse models were employed, including wild-type mice, cholestasis-induced mice, globally farnesoid X-activated receptor (FXR) knock out mice, and high-fat diet-induced (HFD) NAFLD mice. Various administration protocols, spanning one or four weeks and comprising two or three oral doses, were tailored to each model's requirements. Geniposide has positive effects on bile acid and lipid metabolism at doses below 220 mg/kg/day without causing liver injury in normal mice. However, in mice with NAFLD, this dosage is less effective in improving liver function, lipid profiles, and bile acid metabolism compared to lower doses. In cholestasis-induced mice, prolonged use of geniposide at 220 mg/kg/day worsened liver damage. Additionally, in NAFLD mice, this dosage of geniposide for four weeks led to intestinal pyroptosis and liver inflammation. These results highlight the lipid-lowering and bile acid regulatory effects of geniposide, but also warn of potential negative impacts on intestinal epithelial cells, particularly with higher doses and longer treatment durations. Therefore, achieving optimal therapeutic results requires a decrease in treatment duration as the dosage increases, in order to maintain a balanced approach to the use of geniposide in clinical settings.

目的 探讨三七脂肝丸对非酒精性脂肪性肝病(NAFLD)大鼠脂肪酸代谢的影响.方法 大鼠随机分为正常组、模型组、奥贝胆酸组(0.514 mg/kg)和三七脂肝丸低、中、高剂量组(0.77、1.54、3.08 g/kg),每组 8 只.除正常组外,其余各组采用高脂饲料喂养 12 周诱导建立NAFLD大鼠模型.第 9 周,正常组和模型组灌胃给予去离子水,其余各组分别灌胃给予相应剂量药物,连续 4 周,给药结束后(第 12 周末)收集血清和肝、回肠组织样本.生化法检测大鼠血清肝功能、脂质水平及肝组织TG、FFA水平,HE、油红O染色观察肝组织病理学变化,蛋白免疫印迹法检测肝组织FAS、ACC1、SREBP-1c、CD36、SCD-1及回肠组织ZO-1、Occludin蛋白表达.结果 与正常组比较,模型组肝指数、血清ALT、AST、TG、TC、LDL-c、FFA水平、肝组织FFA、TG水平升高(P＜0.05,P＜0.01),血清HDL-c水平降低(P＜0.01),肝组织病理可见脂肪变性,伴有炎性细胞浸润,NAS积分和油红O阳染面积比例升高(P＜0.01),肝组织FAS、ACC1、SREBP-1c、CD36 蛋白表达升高(P＜0.05,P＜0.01),肝组织SCD-1及回肠组织ZO-1、Occludin蛋白表达降低(P＜0.05,P＜0.01);与模型组比较,三七脂肝丸组上述指标均有改善(P＜0.05,P＜0.01),其中高剂量作用更佳.结论 三七脂肝丸能够改善NAFLD大鼠肝功能、血脂水平、肝细胞脂肪变及肠道黏膜机械屏障损伤,其作用机制可能与调节肝脏脂肪酸摄入、脂肪酸从头合成,降低脂肪酸脂毒性的相关蛋白以及肠道紧密连接蛋白表达有关.

The prevalence of nonalcoholic fatty liver disease (NAFLD) has been rising globally. NAFLD patients combined with cholestasis have more obvious liver fibrosis, impaired bile acid (BA), and fatty acid (FA) metabolism and severer liver injury; however, its therapeutic options are limited, and the underlying metabolic mechanisms are understood. Here, we aimed to investigate the effects of farnesoid X receptor (FXR) on BA and FA metabolism in NAFLD combined with cholestasis and related signaling pathways.

Background and aims: The serum metabolites changes in patients with hepatitis B virus (HBV)-related cirrhosis as progression. Peroxisome proliferator-activated receptor gamma (PPARγ) is closely related to lipid metabolism in cirrhotic liver. However, the relationship between fatty acids and the expression of hepatic PPARγ during cirrhosis regression remains unknown. In this study, we explored the serum metabolic characteristics and expression of PPARγ in patients with histological response to treatment with entecavir.Methods: Sixty patients with HBV-related cirrhosis were selected as the training cohort with thirty patients each in the regression (R) group and non-regression (NR) group based on their pathological changes after 48-week treatment with entecavir. Another 72 patients with HBV-related cirrhosis and treated with entecavir were collected as the validation cohort. All of the serum samples were tested using ultra-performance liquid chromatography coupled to tandem mass spectrometry. Data were processed through principal component analysis and orthogonal partial least square discriminant analysis. Hepatic PPARγ expression was observed using immunohistochemistry. The relationship between serum fatty acids and PPARγ was calculated using Pearson’s or Spearman’s correlation analysis.Results: A total of 189 metabolites were identified and 13 differential metabolites were screened. Compared to the non-regression group, the serum level of fatty acids was higher in the R group. At baseline, the expression of PPARγ in hepatic stellate cells was positively correlated with adrenic acid (r2 = 0.451, p = 0.046). The expression of PPARγ in both groups increased after treatment, and the expression of PPARγ in the R group was restored in HSCs much more than that in the NR group (p = 0.042). The adrenic acid and arachidonic acid (AA) in the R group also upgraded more than the NR group after treatment (p = 0.037 and 0.014).Conclusion: Baseline serum differential metabolites, especially fatty acids, were identified in patients with HBV-related cirrhosis patients who achieved cirrhosis regression. Upregulation of adrenic acid and arachidonic acid in serum and re-expression of PPARγ in HSCs may play a crucial role in liver fibrosis improvement.

目的:基于线粒体途径细胞凋亡探讨的茵陈蒿汤和黄芪汤治疗慢性胆汁淤积性肝损伤效应机制的共性和特异性.方法:40只雄性Wistar大鼠随机分为正常组、模型组、茵陈蒿汤组、黄芪汤组,每组10只,除正常组外,其余各组采用α-萘异硫氰酸酯橄榄油溶液灌胃,隔日1次,连续12周,诱导慢性胆汁淤积性肝损伤模型.于第9周首日,给药组分别给予茵陈蒿汤和黄芪汤灌胃,第12周末次给药后收集血清和肝组织样本.检测血清肝功能变化,HE染色观察肝脏组织病理学变化,透射电镜观察肝细胞线粒体形态变化,Western Blot法检测肝组织胆汁酸转运体胆盐输出泵(BSEP)、多药耐药相关蛋白(MRP)2、MRP3以及线粒体途径细胞凋亡相关蛋白细胞色素C(Cyt-C)、半胱氨酸蛋白酶3(Cleaved Caspase-3)、聚腺苷二磷酸核糖聚合酶(Cleaved PARP)、Bax、Bcl-2表达.结果:与正常组比较,模型组大鼠肝组织病理、线粒体形态均发生改变,血清ALT、AST、ALP活性及TBA、TBIL含量升高,肝组织BSEP、MRP2、MRP3、Bcl-2蛋白表达降低,Cyt-C、Cleaved Caspase-3、Cleaved PARP、Bax蛋白表达及Bax/Bcl-2比值升高(P＜0.01);与模型组比较,茵陈蒿汤和黄芪汤均能够改善肝组织病理学和线粒体形态结构,血清ALT、AST活性及TBA、TBIL含量显著降低,肝组织BSEP、MRP2、MRP3、Bcl-2蛋白表达升高,Cyt-C、Cleaved Caspase-3、Cleaved PARP、Bax蛋白表达及Bax/Bcl-2的比值降低(P＜0.01,P＜0.05).与茵陈蒿汤组比较,黄芪汤组肝组织BSEP、MRP2、MRP3、Bcl-2蛋白表达显著升高,肝组织Cyt-C、Cleaved Caspase-3、Cleaved PARP、Bax蛋白表达以及Bax/Bcl-2的比值显著降低(P＜0.05,P＜0.01).结论:茵陈蒿汤和黄芪汤治疗慢性胆汁淤积性肝损伤的作用机制均与调控胆汁酸转运体表达、抑制线粒体途径的细胞凋亡、减轻肝细胞损伤密切相关,黄芪汤调控胆汁酸排泌、抑制线粒体途径细胞凋亡的作用优于茵陈蒿汤.

目的:探究不同剂量和疗程栀子苷调控慢性胆汁淤积的疗效/肝毒性关系,解析其肝保护及肝损伤效应机制.方法:80只Wistar大鼠随机分为正常组,模型组,栀子苷低、中、高剂量(55、110、220 mg/kg)组.采用ANIT灌胃12周诱导慢性胆汁淤积大鼠模型,于第9周给予对应剂量栀子苷灌胃,每日1次,分别于干预7、28 d采集标本.观察肝、回肠组织病理学改变;Western blot法检测血清肝功能及肝组织BSEP、NTCP、MRP3、MRP4、FXR、CAR及回肠组织ZO-1、Occludin蛋白表达.结果:与模型组比较,桅子苷低剂量组给药7、28 d,大鼠血清ALT、AST、TBA水平均降低(P＜0.01,P＜0.05);给药7 d,回肠组织固有层水肿和炎症细胞浸润减轻,回肠组织ZO-1、Occludin和肝组织BSEP蛋白表达升高(P＜0.01);给药28 d,回肠组织ZO-1、Occludin蛋白表达降低(P＜0.05,P＜0.01).栀子苷中、高剂量组给药7、28 d,血清TBIL、DBIL和GGT水平均升高(P＜0.05,P＜0.01),肝、回肠组织病理损伤加重;给药28 d回肠组织ZO-1、Occludin及肝组织BSEP、FXR、CAR蛋白表达降低(P＜0.05,P＜0.01),MRP4蛋白表达升高(P＜0.05,P＜0.01).栀子苷高剂量组给药7、28 d,血清ALB含量均降低(P＜0.01);给药28d,血清ALT、AST活性升高(P＜0.05).结论:不同剂量和疗程栀子苷调控慢性胆汁淤积产生疗效和肝毒性差异的效应机制,与肠黏膜屏障和肝细胞转运功能损伤密切相关.

目的：观察不同剂量野百合碱（MCT）诱导大鼠肝窦阻塞综合征（HSOS）的病理变化，探讨造模剂量-时间致肝损伤的病理特征，并解析其部分作用机制。方法：将72只雄性SD大鼠随机分为正常组（n=12）,MCT低、中、高剂量组（每组n=20）。MCT低、中、高剂量组分别采用MCT 80、120、160 mg·kg-1灌胃一次制备模型，各剂量组大鼠于造模48、120 h处死取材。观测各组大鼠的存活率，检测大鼠体质量、肝质量与血清肝功能变化，采用扫描电镜、苏木素-伊红（HE）染色及天狼星红（SR）染色观察肝组织病理变化，微板法检测肝组织匀浆谷胱甘肽S转移酶（GST）、总超氧化物歧化酶（T-SOD）活性、丙二醛（MDA）含量，实时荧光定量聚合酶链式反应（Real-time PCR）、蛋白免疫印迹法（Western blot）和免疫组化检测肝组织相关指标的表达。结果：与正常组比较，MCT各组血清丙氨酸氨基转移酶/天冬氨酸氨基转移酶（ALT/AST）活性随造模剂量增加而升高（P＜0.05,P＜0.01）；随造模时间增加，MCT低剂量组血清ALT和AST活性均下降（P＜0.01）,MCT中、高剂量组ALT和AST活性均显著升高（P＜0.01）。HE染色显示MCT低、中、高剂量组肝组织肝细胞坏死、炎性细胞浸润及红细胞淤积，电镜下可见肝窦内皮窗孔扩大，“筛状”结构消失，其损伤程度随剂量提高而加重。与正常组比较，MCT低、中、高剂量组肝窦内皮细胞标志物CD44表达减少（P＜0.05,P＜0.01）。SR染色显示造模48 h MCT各组未见阳性染色，120 h MCT各组可见汇管区及肝窦胶原沉积。与正常组比较，MCT各剂量组肝组织MDA含量及GST活性增加，T-SOD活性降低，以MCT中、高剂量组变化显著（P＜0.01）；且造模120 h MCT各组变化呈现剂量依赖性（P＜0.01）。免疫组化及Western blot结果显示，与正常组比较，促炎巨噬细胞标志物CD68蛋白表达随剂量升高而增加（P＜0.01），抗炎巨噬细胞CD163蛋白及mRNA表达随剂量升高而减少（P＜0.01）。与正常组比较，造模后MCT中、高剂量组磷酸化核转录因子-κB/核转录因子-κB（p-NF-κB/NF-κB）、磷酸化蛋白激酶B/蛋白激酶B（p-Akt/Akt）明显升高（P＜0.05,P＜0.01）。MCT造模后肝组织α-平滑肌肌动蛋白（α-SMA）蛋白表达随时间延长、剂量升高增加，α-SMA、Ⅰ型胶原蛋白基因α1（Col 1a1）和Ⅳ型胶原蛋白基因α1（Col 4a1）mRNA表达也随时间延长、剂量升高而明显增加（P＜0.05,P＜0.01）。TUNEL染色结果显示造模后凋亡细胞数量随造模剂量升高而增加，B细胞淋巴瘤-2（Bcl-2）蛋白与Bcl-2相关X蛋白（Bax）比值显著降低（P＜0.01）。结论：不同剂量MCT灌胃诱导的大鼠肝窦阻塞综合征，病变随剂量增加而加重。80 mg·kg-1MCT导致的肝脏损伤可自愈，超过120 mg·kg-1的MCT导致的肝脏损伤会随着时间的延长而加重，甚至出现纤维化、死亡。MCT导致大鼠HSOS的病理机制可能是MCT引发肝组织内强烈的氧化应激，激活促炎巨噬细胞大量分泌炎症因子，进而激活NF-κB/Akt信号通路，导致严重的细胞损伤和死亡。

Nonalcoholic fatty liver disease (NAFLD) is prevalent worldwide, while no pharmaceutical treatment has been approved. Natural herbs are promising for their amelioration effect on lipid metabolism. Theabrownin (TB) and Poria cocos polysaccharide (PCP) have been reported to have effect on hyperlipidemia and diabetes. Here, we compared the effect of individual TB or PCP and the combination of TB and PCP (TB + PCP) on NAFLD phenotypes and the alteration of metabolism in the mice with high-fat diet. The results showed that TB, PCP, and TB + PCP reduced serum and hepatic lipid levels, among which TB + PCP was the most effective. Serum metabolomic profile and liver mRNA analyses revealed that the treatments altered metabolic pathways involved in fatty acid metabolism, bile acid metabolism, and tricarboxylic acid cycle, which was also most significant in the TB + PCP group. This study demonstrated that TB, PCP, especially the combination of TB and PCP could be potential therapeutic formula for NAFLD that promoted lipid utilization and inhibited lipid synthesis and absorption.

目的:观察不同剂量野百合碱(MCT)诱导大鼠肝窦阻塞综合征(HSOS)的病理变化,探讨造模剂量-时间致肝损伤的病理特征,并解析其部分作用机制.方法:将72只雄性SD大鼠随机分为正常组(n=12),MCT低、中、高剂量组(每组n=20).MCT低、中、高剂量组分别采用MCT 80、120、160 mg·kg-1灌胃一次制备模型,各剂量组大鼠于造模48、120 h处死取材.观测各组大鼠的存活率,检测大鼠体质量、肝质量与血清肝功能变化,采用扫描电镜、苏木素-伊红(HE)染色及天狼星红(SR)染色观察肝组织病理变化,微板法检测肝组织匀浆谷胱甘肽S转移酶(GST)、总超氧化物歧化酶(T-SOD)活性、丙二醛(MDA)含量,实时荧光定量聚合酶链式反应(Real-time PCR)、蛋白免疫印迹法(Western blot)和免疫组化检测肝组织相关指标的表达.结果:与正常组比较,MCT各组血清丙氨酸氨基转移酶/天冬氨酸氨基转移酶(ALT/AST)活性随造模剂量增加而升高(P＜0.05,P＜0.01);随造模时间增加,MCT低剂量组血清ALT和AST活性均下降(P＜0.01),MCT中、高剂量组ALT和AST活性均显著升高(P＜0.01).HE染色显示MCT低、中、高剂量组肝组织肝细胞坏死、炎性细胞浸润及红细胞淤积,电镜下可见肝窦内皮窗孔扩大,"筛状"结构消失,其损伤程度随剂量提高而加重.与正常组比较,MCT低、中、高剂量组肝窦内皮细胞标志物CD44表达减少(P＜0.05,P＜0.01).SR染色显示造模48 hMCT各组未见阳性染色,120 hMCT各组可见汇管区及肝窦胶原沉积.与正常组比较,MCT各剂量组肝组织MDA含量及GST活性增加,T-SOD活性降低,以MCT中、高剂量组变化显著(P＜0.01);且造模120 hMCT各组变化呈现剂量依赖性(P＜0.01).免疫组化及Western blot结果显示,与正常组比较,促炎巨噬细胞标志物CD68蛋白表达随剂量升高而增加(P＜0.01),抗炎巨噬细胞CD163蛋白及mRNA表达随剂量升高而减少(P＜0.01).与正常组比较,造模后MCT中、高剂量组磷酸化核转录因子-κB/核转录因子-κB(p-NF-κB/NF-κB)、磷酸化蛋白激酶B/蛋白激酶B(p-Akt/Akt)明显升高(P＜0.05,P＜0.01).MCT造模后肝组织α-平滑肌肌动蛋白(α-SMA)蛋白表达随时间延长、剂量升高增加,α-SMA、Ⅰ型胶原蛋白基因α1(Col 1a1)和Ⅳ型胶原蛋白基因α1(Col 4a1)mRNA表达也随时间延长、剂量升高而明显增加(P＜0.05,P＜0.01).TUNEL染色结果显示造模后凋亡细胞数量随造模剂量升高而增加,B细胞淋巴瘤-2(Bcl-2)蛋白与Bcl-2相关X蛋白(Bax)比值显著降低(P＜0.01).结论:不同剂量MCT灌胃诱导的大鼠肝窦阻塞综合征,病变随剂量增加而加重.80 mg·kg-1 MCT导致的肝脏损伤可自愈,超过120 mg·kg-1 的MCT导致的肝脏损伤会随着时间的延长而加重,甚至出现纤维化、死亡.MCT导致大鼠HSOS的病理机制可能是MCT引发肝组织内强烈的氧化应激,激活促炎巨噬细胞大量分泌炎症因子,进而激活NF-κB/Akt信号通路,导致严重的细胞损伤和死亡.

目的:观察扶正化瘀方对肾纤维化大鼠肾小管上皮细胞间充质转分化(EMT)及TGF-β1/整合素连接激酶(ILK)信号途径关键因子表达的影响,探索其治疗肾纤维化的可能机制.方法:30只雄性SD大鼠随机分为正常组、模型组、扶正化瘀方组,每组10只.以单侧输尿管结扎术诱导单侧输尿管梗阻(UUO)肾纤维化大鼠模型.盐酸水解法检测大鼠肾组织羟脯氨酸(Hyp)含量,天狼猩红染色观察肾组织胶原沉积,免疫组织化学、Western blot和Real-time PCR方法对大鼠肾组织EMT相关指标及TGF-β1/ILK信号途径关键因子蛋白及基因水平进行检测.结果:与正常组比较,模型组大鼠肾组织Hyp含量显著增加(P＜0.01).与模型组比较,扶正化瘀方组大鼠肾组织Hyp含量显著下降(P＜0.01);与正常组比较,模型组肾小管上皮E-cadherin表达显著减少(P＜0.01),而肾小管及间质区α-SMA蛋白表达显著增加(P＜0.01).扶正化瘀方组上调模型组肾小管上皮E-cadherin蛋白表达(P＜0.01),抑制α-SMA蛋白表达(P＜0.01).与正常组比较,模型组TGF-β1、ILK蛋白及其基因水平显著升高(P＜0.01),与模型组比较,扶正化瘀方组TGF-β1、ILK蛋白及其基因水平显著下降(P＜0.01).模型组肾组织p-Akt、p-GSK-3β、β-catenin蛋白水平较正常组显著升高(P＜0.01),扶正化瘀方组显著下调模型组肾组织p-Akt、p-GSK-3β、β-catenin蛋白水平(P＜0.01).模型组肾组织AP-1 mRNA、Cyclin D1 mRNA水平显著高于正常组(P＜0.01),扶正化瘀方组大鼠肾组织AP-1 mRNA水平较模型组显著下调(P＜0.05).结论:扶正化瘀方可通过TGF-β1/ILK信号途径发挥抑制EMT、减轻肾纤维化的作用.

A significant increase of bile acid (BA) levels has been recognized as a general metabolic phenotype of diverse liver diseases. Monitoring of BA profiles has been proposed for etiology differentiation on liver injury. Here, we quantitatively profiled serum BAs of healthy controls and 719 patients with chronic liver disease of five etiologies, hepatitis B virus (HBV), hepatitis C virus (HCV), nonalcoholic steatohepatitis (NASH), alcohol-induced liver disease (ALD), and primary biliary cirrhosis (PBC), and investigated the generality and specificity of different etiologies. The raw data have been deposited into MetaboLights (ID: MTBLS2459). We found that patients with HBV, HCV, and NASH appeared to be more similar, and ALD and PBC patients clustered together. BA profiles, consisting of a total concentration of the 21 quantified BAs [total BAs (TBAs)], 21 BA proportions, and 24 BA relevant variables, were highly different among the etiologies. Specifically, the total BAs was higher in ALD and PBC patients compared with the other three groups. The proportion of conjugated deoxycholates was the highest in HBV-infected patients. The ratio of 12α-hydroxylated (12α-OH) to non-12α-OH BAs was the highest in NASH patients. The proportion of taurine-conjugated BAs was the highest in ALD patients. For PBC patients, the proportion of ursodeoxycholate species was the highest, and the ratio of primary to secondary BAs was the lowest. Comparatively, the difference of BA profiles among cirrhosis patients was consistent but weaker than that of all patients. The correlations between BA profiles and clinical indices were also quite different in different pathological groups, both in all patients and in patients with cirrhosis. Overall, our findings suggested that BA compositions are distinct among patients with different etiologies of chronic liver disease, and some BA-relevant variables are of clinical potentials for liver injury type differentiation, although further validations on more etiologies and populations are needed.