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Objective To screen and verify the Hub genes involved in conjugated bile acid-induced hepatocyte metabolism through transcriptome sequencing analysis. Methods Primary mouse hepatocytes were treated with taurocholic acid (TCA), and the gene expression of TCA-induced hepatocytes was detected by RNA-Seq analysis. Differential expression genes (DEGs) were identified using a fold change ≥1.5 and P value < 0.05 as the screening criteria. The protein-protein interaction (PPI) network of DEGs was constructed using the STRING database, and Hub genes were screened using Cytoscape software based on the PPI network. Real-time qPCR was used to validate the identified Hub gene in vitro and in vivo. Western blotting was employed to detect the expression of proteins related to PSAT1/AKT/GSK3β signaling pathway. The glycogen level in the hepatocytes and liver tissues were detected using glycogen detection kit. Results RNA-Seq analysis identified 293 DEGs in primary mouse hepatocytes upon TCA treatment, including 122 up-regulated genes and 171 down-regulated genes. Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and Gene Set Enrichment Analysis (GSEA) revealed that these DEGs were mainly enriched in metabolism, inflammatory and tumorigenic pathways. Cytohubba plugin identified 16 Hub genes, including Ccnd1, Ppara, Irs1, Ccl2, Ddit3, Serpine1, Nr0b2, Atf3, Phgdh, Trib3, Lepr, G6pc, Psat1, Cxcl2, Asns, and Fgf21. RT-qPCR results demonstrated that various conjugated bile acids, glycocholic acid (GCA), glycochenodeoxycholic acid (GCDCA), and TCA can induce a significant increase in Psat1 mRNA level in primary mouse hepatocytes (P < 0.05). In vivo experiments also showed that the mRNA level of Psat1 was increased significantly in the liver tissues of mouse cholestatic model induced by bile duct ligation (BDL) when compared to the control group (P < 0.05). Western blotting indicated that TCA treatment induced the expression of Psat1 and activation of AKT/GSK3β signaling pathway in primary mouse hepatocytes. TCA treatment also resulted in an increase in glycogen synthesis in primary mouse hepatocytes, which was consistent with an increase in glycogen synthesis in the liver tissue and a decrease in blood glucose level in the mouse model established with BDL (P < 0.05). Conclusion Psat1 may be a gene involved in alteration of glucose metabolism induced by bile acid, and its high expression may be associated with these changes in glucose metabolism in hepatocytes.

Objective To explore the effect and molecular mechanism of artesunate (ART) on cholestatic liver fibrosis induced by 3, 5-diethoxycarbonyl-1, 4-dihydro-2, 4, 6-collidine (DDC). Methods 8-week-old male C57BL/6J mice(n=24) were randomly divided into 3 groups: the control group(n=6), the 0.1% DDC group(n=9) and the 0.1% DDC+ART group(n=9). Mice have been fed with diet containing 0.1% DDC for 2 weeks to establish cholestatic mouse model, and ART was given intramuscular injection (30.0 mg/kg) once a day in the 0.1% DDC+ART group. The indicators of liver injury in serum of mice were detected. HE staining and Sirius red staining were performed, and histopathological scores were obtained according to Scheuer scoring system. RT-qPCR was used to detect the levels of α-SMA, Col1a1, Col1a2 and Tgfβ1 in mouse liver. Hydroxyproline (HYP) content was detected with HYP detection kit. LX2 cells were treated with TGF-β combined with ART, and the proteins related to PI3K/Akt/mTOR pathway were detected by Western blot assay. The expression of α-SMA, hepatic stellate cell (HSC) activation marker, was detected after ART and pathway inhibitors were used to treated with LX2 cells. Results ART significantly ameliorated DDC-induced cholestatic liver injury and liver fibrosis (P < 0.05). ART markedly decreased the levels of ALT, AST, ALP and TBA in serum of mice (P < 0.05). ART inhibited the mRNA expression of α-SMA, Col1a1, Col1a2 and Tgfβ1, and reduced the content of HYP in liver tissues of mice with cholestasis(P < 0.05). In addition, ART obviously inhibited TGF-β-induced activation of PI3K/Akt/mTOR pathway in LX2 cells (P < 0.05). PI3K/Akt/mTOR signaling pathway inhibitors enhanced the inhibitory effect of HSC activation induced by TGF-β(P < 0.05). Conclusion ART observably improves DDC-induced cholestatic liver fibrosis, and it can inhibit HSC activation by regulating PI3K/Akt/mTOR signaling pathway, thus exerting anti-fibrosis effect.

Objective To investigate the underlying molecular mechanism by which melatonin suppresses bile acids (BAs)-initiated expression of chemokine C-X-C motif chemokine ligand 2 (Cxcl2). Methods Mice were fed a 1% cholic acid (CA)-supplemented diet for 2 weeks to establish a model of cholestatic disease, and were given an intraperitoneal injection of melatonin (100 mg/kg) once a day for 2 weeks. Then, the liver tissues were collected for HE staining, and the liver histology was scored using the Scheuer scoring system. Effects of melatonin on the expression of Cxcl2, Cxcr2, myeloperoxidase (Mpo) and c-Jun in the liver tissues at mRNA and protein levels were detected by real-time qPCR and Western blotting. The expression of c-Jun in the liver tissues was measured by immunohistochemical assay. Primary mouse hepatocytes were isolated and cultured, and then treated with taurine-conjugated bile acid, taurocholic acid (TCA) and melatonin in order to explore the mechanism of melatonin reducing BAs-initiated Cxcl2 expression. Furthermore, these primarily cultured cells were treated with anisomycin (JNK agonist) and melatonin to further detect the expression of Cxcl2. Results Melatonin treatment significantly ameliorated cholestasis-induced inflammatory liver injury (P < 0.05), and reduced the elevated expression of Cxcl2, Cxcr2, Mpo(a biomarker of neutrophils) and the transcription factor c-Jun in the livers of cholestatic disease mice (P

Grain boundary segmentation is crucial for the quantitative analysis of grain structures and material optimization. However, challenges persist due to noise interference, high labeling costs, and low detection Accuracy. Therefore, we propose a semi-supervised method called Semi-SRUnet, which is based on teacher–student knowledge distillation and pseudo-label repair to achieve grain boundary detection for a small number of labels. Specifically, the method introduces SCConv (Spatial and Channel Reconstruction Convolution) and boundary regression to improve the U-Net (a convolutional neural network architecture) as a teacher network. These innovations aim to reduce spatial and channel redundancy, expand the receptive field, and effectively capture contextual information from images, thereby improving feature extraction robustness and boundary precision in noisy environments. Additionally, we designed a pseudo-label repair algorithm to enhance the Accuracy of pseudo-labels generated by the teacher network and used knowledge distillation to train a lightweight student network. The experimental results demonstrate that Semi-SRUnet achieves 88.86% mean Intersection over Union (mIoU), 96.64% mean Recall (mRecall), 91.5% mean Precision (mPrecision), and 98.77% Accuracy, surpassing state-of-the-art models and offering a novel approach for reliable grain boundary segmentation and analysis. [ABSTRACT FROM AUTHOR]

Internal pore defects are inevitable during laser powder bed fusion (LPBF), which have a significant impact on the mechanical properties of the parts. Therefore, detecting pores and obtaining their morphology will contribute to the quality of LPBF parts. Currently, supervised models are used for defect image detection, which requires a large amount of LPBF sample data, image labeling, and computing power equipment during the training process, resulting in high detection costs. This study extensively collected LPBF sample data and proposed a method for pore defect classification by obtaining its morphological features while detecting pore defects in optical microscopy (OM) images under various conditions. Compared with other advanced models, the proposed method achieves better detection accuracy on pore defect datasets with limited data. In addition, quickly detecting pore defects in a large number of labeling ground truth images will also contribute to the development of deep learning. In terms of image segmentation, the average accuracy scores of this method in the test images exceed 85%. The research results indicate that the algorithm proposed in this paper is suitable for quickly and accurately identifying pore defects from optical microscopy images. [ABSTRACT FROM AUTHOR]

Detecting posture changes of athletes in sports is an important task in teaching and training competitions, but its detection remains challenging due to the diversity and complexity of sports postures. This paper introduces a single-stage pose estimation algorithm named yolov8-sp. This algorithm enhances the original yolov8 architecture by incorporating the concept of multi-dimensional feature fusion and the attention mechanism for automatically capturing feature importance. Furthermore, in this paper, angle extraction is conducted for three crucial motion joints in the motion scene, with polynomial corrections applied across successive frames. In comparison with the baseline yolov8, the improved model significantly outperforms it in A P 50 (average precision) aspects. Specifically, the model's performance improves from 84.5 AP to 87.1 AP, and the performance of A P 50 – 95 , A P M , and A P L aspects also shows varying degrees of improvement; the joint angle detection accuracy under different sports scenarios is tested, and the overall accuracy is improved from 73.2% to 89.0%, which proves the feasibility of the method for posture estimation of the human body in sports and provides a reliable tool for the analysis of athletes' joint angles. [ABSTRACT FROM AUTHOR]

Metal FDM technology overcomes the problems of high cost, high energy consumption and high material requirements of traditional metal additive manufacturing by combining FDM and powder metallurgy and realizes the low-cost manufacturing of complex metal parts. In this work, 15-5PH stainless steel granules with a powder content of 90% and suitable for metal FDM were developed. The flowability and formability of the feedstock were investigated and the parts were printed. A two-step (solvent and thermal) debinding process is used to remove the binder from the green part. After being kept at 75 °C in cyclohexane for 24 h, the solvent debinding rate reached 98.7%. Following thermal debinding, the material's weight decreased by slightly more than 10%. Sintering was conducted at 1300 °C, 1375 °C and 1390 °C in a hydrogen atmosphere. The results show that the shrinkage of the sintered components in the X-Y-Z direction remains quite consistent, with values ranging from 13.26% to 19.58% between 1300 °C and 1390 °C. After sintering at 1390 °C, the material exhibited a relative density of 95.83%, a hardness of 101.63 HRBW and a remarkable tensile strength of 770 MPa. This work realizes the production of metal parts using 15-5PH granules' extrusion additive manufacturing, providing a method for the low-cost preparation of metal parts. And it provides a useful reference for the debinding and sintering process settings of metal FDM. In addition, it also enriches the selection range of materials for metal FDM. [ABSTRACT FROM AUTHOR]

Background & Aims: There is an unmet clinical need for non‐invasive tests to diagnose non‐alcoholic fatty liver disease (NAFLD) and individual fibrosis stages. We aimed to test whether urine protein panels could be used to identify NAFLD, NAFLD with fibrosis (stage F ≥ 1) and NAFLD with significant fibrosis (stage F ≥ 2). Methods: We collected urine samples from 100 patients with biopsy‐confirmed NAFLD and 40 healthy volunteers, and proteomics and bioinformatics analyses were performed in this derivation cohort. Diagnostic models were developed for detecting NAFLD (UPNAFLD model), NAFLD with fibrosis (UPfibrosis model), or NAFLD with significant fibrosis (UPsignificant fibrosis model). Subsequently, the derivation cohort was divided into training and testing sets to evaluate the efficacy of these diagnostic models. Finally, in a separate independent validation cohort of 100 patients with biopsy‐confirmed NAFLD and 45 healthy controls, urinary enzyme‐linked immunosorbent assay analyses were undertaken to validate the accuracy of these new diagnostic models. Results: The UPfibrosis model and the UPsignificant fibrosis model showed an AUROC of.863 (95% CI:.725–1.000) and 0.858 (95% CI:.712–1.000) in the training set; and.837 (95% CI:.711–.963) and.916 (95% CI:.825–1.000) in the testing set respectively. The UPNAFLD model showed an excellent diagnostic performance and the area under the receiver operator characteristic curve (AUROC) exceeded.90 in the derivation cohort. In the independent validation cohort, the AUROC for all three of the above diagnostic models exceeded.80. Conclusions: Our newly developed models constructed from urine protein biomarkers have good accuracy for non‐invasively diagnosing liver fibrosis in NAFLD. [ABSTRACT FROM AUTHOR]

Bile components play a critical role in maintaining gut microbiota homeostasis. In cholestasis, bile secretion is impaired, leading to liver injury. However, it remains to be elucidated whether gut microbiota plays a role in cholestatic liver injury. Here, we performed a sham operation and bile duct ligation (BDL) in antibiotic-induced microbiome depleted (AIMD) mice and assessed liver injury and fecal microbiota composition in these mice. Significant reductions in gut microbiota richness and diversity were found in AIMD-sham mice when compared to sham controls. Three-day BDL leads to great elevation of plasma ALT, ALP, total bile acids, and bilirubin where reduced diversity of the gut microbiota was also found. AIMD further aggravated cholestatic liver injury evidenced by significantly higher levels of plasma ALT and ALP, associated with further reduced diversity and increased Gram-negative bacteria in gut microbiota. Further analyses revealed increased levels of LPS in the plasma of AIMD-BDL mice where elevated expression of inflammatory genes and decreased expression of hepatic detoxification enzymes were also found in liver when compared to the BDL group. These findings indicate that gut microbiota plays a critical role in cholestatic liver injury. Maintaining its homeostasis may alleviate liver injury in patients with cholestasis. [ABSTRACT FROM AUTHOR]

Cholangiocytes play a crucial role in bile formation. Cholangiocyte injury causes cholestasis, including primary biliary cholangitis (PBC). However, the etiology of PBC remains unclear despite being characterized as an autoimmune disease. Using single-cell RNA sequencing (scRNA-seq), fluorescence-activated-cell-sorting, multiplex immunofluorescence (IF) and RNAscope analyses, we identified unique DUOX2+ACE2+ small cholangiocytes in human and mouse livers. Their selective decrease in PBC patients was associated with the severity of disease. Moreover, proteomics, scRNA-seq, and qPCR analyses indicated that polymeric immunoglobulin receptor (pIgR) was highly expressed in DUOX2+ACE2+ cholangiocytes. Serum anti-pIgR autoantibody levels were significantly increased in PBC patients, regardless of positive and negative AMA-M2. Spatial transcriptomics and multiplex IF revealed that CD27+ memory B and plasma cells accumulated in the hepatic portal tracts of PBC patients. Collectively, DUOX2+ACE2+ small cholangiocytes are pathogenic targets in PBC, and preservation of DUOX2+ACE2+ cholangiocytes and targeting anti-pIgR autoantibodies may be valuable strategies for therapeutic interventions in PBC. The aetiology of primary biliary cholangitis (PBC) remains unclear. Here, the authors find that the numbers of DUOX2 + ACE2 + small cholangiocytes in human and mouse livers are inversely associated with disease severity, and present data indicating that they may be the target of polymeric immunoglobulin receptor (pIgR) -mediated humoral responses, suggesting that preservation of these cells and targeting anti-pIgR autoantibodies may be valuable strategies for therapeutic interventions in PBC. [ABSTRACT FROM AUTHOR]

In this article, molecular dynamics simulation is used to primarily explore the lattice matching problem of solute atoms in different configurations after entering the Fe lattice at 0 K temperature. It reveals that by changing the initial configurations, metal solute atoms (Cr, Cu, Ti, Al) are more stable in the <110> configuration while C atom tends to stay in the octahedral gap of the BCC lattice. In addition, the matching mechanisms of different solute atoms in BCC Fe lattice in different configurations are elaborated in detail. The relaxation of metal solute atoms in different configurations eventually forms different stable structures while C atoms only exist in the nearest‐neighbor octahedral gap of the BCC Fe lattice in each configuration. [ABSTRACT FROM AUTHOR]

Semaphorin 7A (SEMA7A) is a membrane‐bound protein that involves axon growth and other biological processes. SEMA7A mutations are associated with vertebral fracture and Kallmann syndrome. Here, we report a case with a mutation in SEMA7A that displays familial cholestasis. WGS reveals a SEMA7AR148W homozygous mutation in a female child with elevated levels of serum ALT, AST, and total bile acid (TBA) of unknown etiology. This patient also carried a SLC10A1S267F allele, but Slc10a1S267F homozygous mice exhibited normal liver function. Similar to the child, Sema7aR145W homozygous mice displayed elevated levels of serum ALT, AST, and TBA. Remarkably, liver histology and LC‐MS/MS analyses exhibited hepatocyte hydropic degeneration and increased liver bile acid (BA) levels in Sema7aR145W homozygous mice. Further mechanistic studies demonstrated that Sema7aR145W mutation reduced the expression of canalicular membrane BA transporters, bile salt export pump (Bsep), and multidrug resistance‐associated protein‐2 (Mrp2), causing intrahepatic cholestasis in mice. Administration with ursodeoxycholic acid and a dietary supplement glutathione improved liver function in the child. Therefore, Sema7aR145W homozygous mutation causes intrahepatic cholestasis by reducing hepatic Bsep and Mrp2 expression. SYNOPSIS: A new type of progressive familial intrahepatic cholestasis (PFIC) was caused by the homozygous R148W mutation in SEMA7A. Preliminary mechanistic studies revealed that the mutation reduced hepatic expression of canalicular membrane bile acid (BA) efflux transporters Bsep and Mrp2, resulting in intrahepatic cholestasis. A female child patient presented with elevated levels of serum ALT, AST, and total bile acid (TBA) of unknown etiology.A SEMA7AR148W homozygous mutation was identified in the child patient.The Sema7aR145W (human R148W) homozygous mice displayed elevated levels of serum ALT, AST, and TBA, as well as hepatocyte hydropic degeneration and intrahepatic accumulation of conjugated BAs.The Sema7aR145W homozygous mutation reduced the expression of canalicular membrane Bsep and Mrp2 in mouse livers and sandwich‐cultured primary hepatocytes.Administration with ursodeoxycholic acid (UDCA) and a dietary supplement glutathione (GSH) were effective for this new type of PFIC. [ABSTRACT FROM AUTHOR]

We report here, for the first time, that interleukin 32 (IL32) expression is elevated in the liver of patients with obstructive cholestasis, and its upregulation ameliorates cholestatic liver injury by repressing bile acid (BA) synthesis and the inflammatory response. Cholestasis is characterized by excessive accumulation of intrahepatic BA that causes liver injury and inflammation.1,2 IL32, a predominantly intracellular proinflammatory mediator, is involved in infectious diseases and cancers,3-7 but at present, its role in cholestasis remains unknown. To reveal how IL32 represses BA synthesis in cholestasis, we assessed the expression of major genes involved in the biosynthesis and transportation of BA. Together, these data indicate that hepatic IL32 can ameliorate cholestatic liver injury by repressing BA synthesis and liver inflammation. [Extracted from the article]

Background: Renal tubulointerstitial fibrosis (TIF) is commonly the final result of a variety of progressive injuries and leads to end-stage renal disease. There are few therapeutic agents currently available for retarding the development of renal TIF.Purpose: The aim of the present study is to evaluate the role of arctigenin (ATG), a lignan component derived from dried burdock (Arctium lappa L.) fruits, in protecting the kidney against injury by unilateral ureteral obstruction (UUO) in rats.Methods: Rats were subjected to UUO and then administered with vehicle, ATG (1 and 3mg/kg/d), or losartan (20mg/kg/d) for 11 consecutive days. The renoprotective effects of ATG were evaluated by histological examination and multiple biochemical assays.Results: Our results suggest that ATG significantly protected the kidney from injury by reducing tubular dilatation, epithelial atrophy, collagen deposition, and tubulointerstitial compartment expansion. ATG administration dramatically decreased macrophage (CD68-positive cell) infiltration. Meanwhile, ATG down-regulated the mRNA levels of pro-inflammatory chemokine monocyte chemoattractant protein-1 (MCP-1) and cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interferon-γ (IFN-γ), in the obstructed kidneys. This was associated with decreased activation of nuclear factor κB (NF-κB). ATG attenuated UUO-induced oxidative stress by increasing the activity of renal manganese superoxide dismutase (SOD2), leading to reduced levels of lipid peroxidation. Furthermore, ATG inhibited the epithelial-mesenchymal transition (EMT) of renal tubules by reducing the abundance of transforming growth factor-β1 (TGF-β1) and its type I receptor, suppressing Smad2/3 phosphorylation and nuclear translocation, and up-regulating Smad7 expression. Notably, the efficacy of ATG in renal protection was comparable or even superior to losartan.Conclusion: ATG could protect the kidney from UUO-induced injury and fibrogenesis by suppressing inflammation, oxidative stress, and tubular EMT, thus supporting the potential role of ATG in renal fibrosis treatment. [ABSTRACT FROM AUTHOR]

Abstract: Linear discriminant analysis (LDA) often suffers from the small sample size problem when dealing with high-dimensional face data. Random subspace can effectively solve this problem by random sampling on face features. However, it remains a problem how to construct an optimal random subspace for discriminant analysis and perform the most efficient discriminant analysis on the constructed random subspace. In this paper, we propose a novel framework, random discriminant analysis (RDA), to handle this problem. Under the most suitable situation of the principal subspace, the optimal reduced dimension of the face sample is discovered to construct a random subspace where all the discriminative information in the face space is distributed in the two principal subspaces of the within-class and between-class matrices. Then we apply Fisherface and direct LDA, respectively, to the two principal subspaces for simultaneous discriminant analysis. The two sets of discriminant analysis features from dual principal subspaces are first combined at the feature level, and then all the random subspaces are further integrated at the decision level. With the discriminating information fusion at the two levels, our method can take full advantage of useful discriminant information in the face space. Extensive experiments on different face databases demonstrate its performance. [Copyright &y& Elsevier]

As a central regulator for endoplasmic reticulum (ER) stress, glucose-regulated protein 78 (GRP78), controls the activation of ER-transmembrane signaling mechanisms by inducing unfolded protein response (UPR) in response to ER stress. Although limited glucose availability often occurs in poorly vascularized solid cancers, cancer cells often initiate the UPR to support cellular homeostasis and survival under stress conditions. Therefore, targeting GRP78 expression and UPR pathway activation may provide a new strategy for anticancer therapy. Based on this premise, we investigated the molecular mechanisms of a synthetic quinolone derivative, 2-hexyl-3-methyl-4(1H)-quinolinone (HHQ-4), in regulating the GRP78 expression and UPR transcriptional program under glucose deprivation or 2-deoxy- d -glucose (2-DG)-stressed conditions. We found that HHQ-4 suppressed the transcriptional and translational expression of GRP78 gene in glucose-deprived breast cancer cells. HHQ-4 also showed selective antiproliferative activity against glucose-deprived breast cancer cells. Constitutive expression of GRP78 completely prevented breast cancer cells from HHQ-4-mediated proliferation inhibition during glucose starvation, stressing the important role of suppression of the GRP78 in HHQ-4-mediated cell proliferation inhibition. HHQ-4 was also found to exert inhibitory activity against breast cancer cell proliferation by suppressing three survival arms of the UPR, including PERK/eIF2α/ATF4, IRE1/XBP1, and ATF6, which orchestrate an intricate signaling network to modulate GRP78 gene transcription under glucose-deprived stress. Furthermore, HHQ-4 combined with 2-DG synergistically inhibited breast cancer cell proliferation. Our findings show HHQ-4 could be a promising candidate, alone or in combination with 2-DG, for selectively inhibiting breast cancer cell proliferation by down-regulating the transcription and expression of GRP78 under stressful microenvironments. Unlabelled Image • HHQ-4 acts as a GRP78 down-regulator during glucose deprivation. • HHQ-4 shows antiproliferative activity against glucose-deprived breast cancer cells. • Overexpressing GRP78 prevents HHQ-4-induced cell proliferation inhibition. • Disruption of UPR signaling is responsible for down-regulation of GRP78 by HHQ-4. [ABSTRACT FROM AUTHOR]

Background and Aims: Organic anion-transporting polypeptides (OATPs) play a crucial role in the transport of bile acids and bilirubin. In our previous study, interleukin 6 (IL-6) reduced OATP1B3 levels in cholestatic disease. However, it remains unclear whether IL-6 inhibits OATP1B1 expression in cholestatic diseases. This study aimed to investigate whether IL-6 can inhibit OATP1B1 expression and explore the underlying mechanisms., Methods: The effect of stimulator of interferon genes (STING) signaling on inflammatory factors was investigated in a cholestatic mouse model using RT-qPCR and enzyme-linked immunosorbent assay. To assess the impact of inflammatory factors on OATP1B1 expression in hepatocellular carcinoma, we analyzed OATP1B1 expression by RT-qPCR and Western Blot after treating PLC/PRF/5 cells with TNF-α, IL-1β, and IL-6. To elucidate the mechanism by which IL-6 inhibits OATP1B1 expression, we examined the expression of the OATP1B1 regulator TCF4 in PLC/PRF/5 and HepG2 cells using RT-qPCR and Western Blot. The interaction mechanism between β-catenin/TCF4 and OATP1B1 was investigated by knocking down β-catenin/TCF4 through siRNA transfection., Results: The STING inhibitor decreased inflammatory factor levels in the cholestatic mouse model, with IL-6 exhibiting the most potent inhibitory effect on OATP1B1. IL-6 downregulated β-catenin/TCF4, leading to decreased OATP1B1 expression. Knocking-down β-catenin/TCF4 counteracted the β-catenin/TCF4-mediated repression of OATP1B1., Conclusions: STING-mediated IL-6 up-regulation may inhibit OATP1B1, leading to reduced transport of bile acids and bilirubin by OATP1B1. This may contribute to altered pharmacokinetics in patients with diseases associated with increased IL-6 production., Competing Interests: The authors have no conflict of interests related to this publication., (© 2024 Authors.)

Background and Aims: Inflammatory response is crucial for bile acid (BA)-induced cholestatic liver injury, but molecular mechanisms remain to be elucidated. Solute Carrier Family 35 Member C1 (SLC35C1) can transport Guanosine diphosphate-fucose into the Golgi to facilitate protein glycosylation. Its mutation leads to the deficiency of leukocyte adhesion and enhances inflammation in humans. However, little is known about its role in liver diseases., Approach and Results: Hepatic SLC35C1 mRNA transcripts and protein expression were significantly increased in patients with obstructive cholestasis and mouse models of cholestasis. Immunofluorescence revealed that the upregulated SLC35C1 expression mainly occurred in hepatocytes. Liver-specific ablation of Slc35c1 ( Slc35c1 cKO ) significantly aggravated liver injury in mouse models of cholestasis induced by bile duct ligation and 1% cholic acid-feeding, evidenced by increased liver necrosis, inflammation, fibrosis, and bile ductular proliferation. The Slc35c1 cKO increased hepatic chemokine Ccl2 and Cxcl2 expression and T cell, neutrophil, and F4/80 macrophage infiltration but did not affect the levels of serum and liver BA in mouse models of cholestasis. Liquid chromatography with tandem mass spectrometry analysis revealed that hepatic Slc35c1 deficiency substantially reduced the fucosylation of cell-cell adhesion protein CEACAM1 at N153. Mechanistically, cholestatic levels of conjugated BAs stimulated SLC35C1 expression by activating the STAT3 signaling to facilitate CEACAM1 fucosylation at N153, and deficiency in the fucosylation of CEACAM1 at N135 enhanced the BA-stimulated CCL2 and CXCL2 mRNA expression in primary mouse hepatocytes and Primary Liver Carcinoma/Poliomyelitis Research Foundation/5- ASBT cells., Conclusions: Elevated hepatic SLC35C1 expression attenuates cholestatic liver injury by enhancing CEACAM1 fucosylation to suppress CCL2 and CXCL2 expression and liver inflammation., (Copyright © 2024 The Author(s). Published by Wolters Kluwer Health, Inc.)

Background and Aims: Recent studies reported that the hepatic expression of AQP8 and AQP9 was downregulated in bile duct-ligated (BDL) rats and that overexpression of human AQP1 in the rat liver attenuated cholestasis. However, the hepatic expression of AQP10 and its regulatory mechanism in human cholestasis remain unclear., Methods: Serum and liver samples were collected from 34 patients with obstructive cholestasis and from 12 control patients. Eight-week-old male C57BL/6J mice were intravenously injected with an adeno-associated virus 8 (AAV8) encoding human AQP10 driven by a hepatocyte-specific Alb promotor (AAV8- Alb promotor- hAQP10 ) for functional studies. Constructs of the AQP10 promoter and PLC/PRF/5-ASBT cell lines were used for regulatory mechanism studies., Results: AQP10 was significantly downregulated in patients with obstructive cholestasis and negatively associated with the serum levels of total bile acid (TBA). The hepatocyte-specific overexpression of  hAQP10  significantly attenuated the cholestatic liver injury and intrahepatic bile acids (BA) accumulation in BDL mice. Conjugated BAs, such as TCA and inflammatory factor TNFα, significantly repressed AQP10 expression. Furthermore, NFκB p65/p50 directly bound to the AQP10 promotor and decreased its activity in PLC/RPF/5- ASBT  cells and in the livers of patients with obstructive cholestasis. However, these changes were diminished by BAY 11-7082 (a specific inhibitor of NFκB signaling)., Conclusion: We are the first to report that AQP10 was significantly decreased in patients with obstructive cholestasis. AQP10 overexpression significantly attenuated cholestatic liver injury in BDL mice. Therefore, overexpression of hAQP10 in the liver may be a valuable strategy for cholestasis intervention., (© 2023 The Authors.)

Genetic polymorphisms are associated with the development of nonalcoholic fatty liver disease (NAFLD). Semaphorin7a (Sema7a) deficiency in mouse peritoneal macrophages reduces fatty acid (FA) oxidation. Here, we identified 17 individuals with SEMA7A heterozygous mutations in 470 patients with biopsy-proven NAFLD. SEMA7A heterozygous mutations increased susceptibility to NAFLD, steatosis severity, and NAFLD activity scores in humans and mice. The Sema7aR145W mutation (equivalent to human SEMA7AR148W) significantly induced small lipid droplet accumulation in mouse livers compared with WT mouse livers. Mechanistically, the Sema7aR145W mutation increased N-glycosylated Sema7a and its receptor integrin β1 proteins in the cell membranes of hepatocytes. Furthermore, Sema7aR145W mutation enhanced its protein interaction with integrin β1 and PKC-α and increased PKC-α phosphorylation, which were both abrogated by integrin β1 silencing. Induction of PKCα&#95;WT, but not PKCα&#95;dominant negative, overexpression induced transcriptional factors Srebp1, Chrebp, and Lxr expression and their downstream Acc1, Fasn, and Cd36 expression in primary mouse hepatocytes. Collectively, our findings demonstrate that the SEMA7AR148W mutation is a potentially new strong genetic determinant of NAFLD and promotes intrahepatic lipid accumulation and NAFLD in mice by enhancing PKC-α-stimulated FA and triglyceride synthesis and FA uptake. The inhibition of hepatic PKC-α signaling may lead to novel NAFLD therapies.

Cholestasis is a common condition in which the flow of bile from the liver to the intestines is inhibited. It has been shown that organic anion-transporting polypeptide 3A1 (OATP3A1) is upregulated in cholestasis to promote bile acid efflux transport. We have previously shown that the growth factor fibroblast growth factor 19 and inflammatory mediator tumor necrosis factor α (TNFα) increased OATP3A1 mRNA levels in hepatoma peritoneal lavage cell/PRF/5 cell lines. However, the mechanism underlying TNFα-stimulated OATP3A1 expression in cholestasis is unknown. To address this, we collected plasma samples from control and obstructive cholestasis patients and used ELISA to detect TNFα levels. We found that the TNFα levels of plasma and hepatic mRNA transcripts were significantly increased in obstructive cholestatic patients relative to control patients. A significant positive correlation was also observed between plasma TNFα and liver OATP3A1 mRNA transcripts in patients with obstructive cholestasis. Further mechanism analysis revealed that recombinant TNFα induced OATP3A1 expression and activated NF-κB and extracellular signal-regulated kinase (ERK) signaling pathways as well as expression of related transcription factors p65 and specificity protein 1 (SP1). Dual-luciferase reporter and chromatin immunoprecipitation assays showed that recombinant TNFα upregulated the binding activities of NF-κB p65 and SP1 to the OATP3A1 promoter in peritoneal lavage cell/PRF/5 cells. These effects were diminished following the application of NF-κB and ERK inhibitors BAY11-7082 and PD98059. We conclude that TNFα stimulates hepatic OATP3A1 expression in human obstructive cholestasis by activating NF-κB p65 and ERK-SP1 signaling. These results suggest that TNFα-activated NF-κB p65 and ERK-SP1 signaling may be a potential target to ameliorate cholestasis-associated liver injury., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)