Betaine/GABA transporter-1 (BGT-1) deficiency in mouse prevents acute liver failure in vivo and hepatocytes apoptosis in vitro

Mesenchymal stem cells (MSCs) and hepatocytes are two attractive sources of cell-based therapies for acute liver failure (ALF). The cotransplantation of hepatocytes with MSCs can improve the therapeutic performance for the treatment of ALF. However, the therapeutic potential of conditioned medium (CM) derived from MSCs cocultured with hepatocytes (MSC-H-CM) remains unclear. The purpose of this study was to investigate the effects of MSC-H-CM on damaged hepatocytes in vitro and on D-galactosamine-induced ALF in vivo. D-Galactosamine-treated L02 cells cultured in MSC-H-CM exhibited higher of cell viability and total protein synthesis than L02 cells cultured in MSC-CM, CM derived from hepatocytes (H-CM), MSC-CM + H-CM, or with nonconditioned medium (NCM). Lactate dehydrogenase and aspartate aminotransferase levels were lower in the supernatant of damaged L02 cells cultured in MSC-H-CM than in that of L02 cells cultured in other types of CM. The lowest percentage of apoptotic cells was observed after the MSC-H-CM treatment. When CM was injected into the tail vein of rats with ALF, MSC-H-CM was the most successful at preventing the release of liver injury biomarkers and in promoting the recovery of liver structure. The greatest survival rate 7 days after the first treatment was observed in the MSC-H-CM-treated rats. Our results reveal that the delivery of MSC-H-CM could be a novel strategy for integrating the therapeutic potentials of hepatocytes and MSCs for the treatment of ALF.

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Fusobacterium Nucleatum Promotes the Development of Acute Liver Failure by Inhibiting the NAD+ Salvage Metabolic Pathway

10.21203/rs.3.rs-1091141/v1 ◽

2021 ◽

Author(s):

Pan Cao ◽

Qian Chen ◽

Chunxia Shi ◽

Luwen Wang ◽

Zuojiong Gong

Keyword(s):

Energy Metabolism ◽

Liver Failure ◽

Acute Liver Failure ◽

Fusobacterium Nucleatum ◽

Mucosal Barrier ◽

Liver Inflammation ◽

Ampk Signaling ◽

Inflammatory Damage

Abstract Background: Acute liver failure (ALF) patients are often accompanied by severe energy metabolism abnormalities and intestinal microecological imbalance. The intestinal mucosal barrier is severely damaged. Intestinal endotoxin can induce intestinal endotoxemia through the "Gut-Liver axis". More and more evidence shows that members of the gut microbiota, especially Fusobacterium nucleatum (F. nucleatum), are related to inflammatory bowel disease, but whether F. nucleatum is involved in the development of ALF and whether it affects the liver energy metabolism is unclear. Methods: This study first detected the abundance of F. nucleatum and its effect on ALF disease, and explored whether F. nucleatum aggravated liver inflammation in vitro and in vivo. Results: Our data showed that liver tissues of ALF patients contained different abundances of F. nucleatum, which were related to the degree of liver inflammation. In addition, we found that F. nucleatum infection affected the energy metabolism of the liver during the development of ALF, inhibited the synthesis pathway of nicotinamide adenine dinucleotide (NAD+)'s salvage metabolism, and promoted inflammatory damage in the liver. In terms of mechanism, F. nucleatum inhibited NAD+ and the NAD+-dependent SIRT1/AMPK signaling pathway, and promoted liver damage of ALF. Conclusions: F. nucleatum coordinates a molecular network including NAD+ and SIRT1 to control the progress of ALF. Detection and targeting of F. nucleatum and its related pathways may provide valuable insights for the treatment of ALF.

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HBx facilitates ferroptosis in acute liver failure via EZH2 mediated SLC7A11 suppression

Journal of Biomedical Science ◽

10.1186/s12929-021-00762-2 ◽

2021 ◽

Vol 28 (1) ◽

Author(s):

Guo-Zhen Liu ◽

Xu-Wen Xu ◽

Shu-Hui Tao ◽

Ming-Jian Gao ◽

Zhou-Hua Hou

Keyword(s):

Liver Failure ◽

Acute Liver Failure ◽

Epigenetic Modification ◽

Hbv Infection ◽

High Rate ◽

Mice Model ◽

Primary Hepatocytes ◽

Hepatocyte Injury

Abstract Background Acute liver failure (ALF) is a syndrome of severe hepatocyte injury with high rate of mortality. Hepatitis B virus (HBV) infection is the major cause of ALF worldwide, however, the underlying mechanism by which HBV infection leads to ALF has not been fully disclosed. Methods D-GalN-induced hepatocyte injury model and LPS/D-GalN-induced ALF mice model were used to investigate the effects of HBV X protein (HBx) in vitro and in vivo, respectively. Cell viability and the levels of Glutathione (GSH), malondialdehyde (MDA) and iron were measured using commercial kits. The expression of ferroptosis-related molecules were detected by qRT-PCR and western blotting. Epigenetic modification and protein interaction were detected by chromatin immunoprecipitation (ChIP) assay and co-immunoprecipitation (co-IP), respectively. Mouse liver function was assessed by measuring aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The histological changes in liver tissues were monitored by hematoxylin and eosin (H&E) staining, and SLC7A11 immunoreactivity was assessed by immunohistochemistry (IHC) analysis. Results D-GalN triggered ferroptosis in primary hepatocytes. HBx potentiated D-GalN-induced hepatotoxicity and ferroptosis in vitro, and it suppressed SLC7A11 expression through H3K27me3 modification by EZH2. In addition, EZH2 inhibition or SLC7A11 overexpression attenuated the effects of HBx on D-GalN-induced ferroptosis in primary hepatocytes. The ferroptosis inhibitor ferrostatin-1 (Fer-1) protected against ALF and ferroptosis in vivo. By contrast, HBx exacerbates LPS/D-GalN-induced ALF and ferroptosis in HBx transgenic (HBx-Tg) mice. Conclusion HBx facilitates ferroptosis in ALF via EZH2/H3K27me3-mediated SLC7A11 suppression.

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CCR2-Overexpressing Mesenchymal Stem Cells Targeting Damaged Liver Enhances Recovery of Acute Liver Failure

10.21203/rs.3.rs-581837/v1 ◽

2021 ◽

Author(s):

Ruixuan Xu ◽

Jiarou Shan ◽

Beibei Ni ◽

Lijie Pan ◽

Guo Lv ◽

...

Keyword(s):

Liver Failure ◽

Acute Liver Failure ◽

Chemokine Receptors ◽

Near Infrared ◽

Ex Vivo ◽

Inflammatory Infiltration ◽

In Vitro Expansion ◽

Novel Strategy

Abstract Background: Mesenchymal stem cell (MSC) transplantation is emerging as a promising cell therapeutic strategy in acute liver failure (ALF) clinical research. The potency of MSCs to migrate and engraft into targeted lesions could largely determine their clinical efficacy, in which chemokine/receptor axes play a crucial role. Unfortunately, the downregulation of chemokine receptors expression after in vitro expansion results in a poor homing capacity of MSCs.Methods: By evaluating the chemokine expression profile in the liver of ALF patients and ALF mice, we found that CCL2 expression was highly upregulated in damaged livers, while the corresponding receptor, CCR2, was lacking in cultured MSCs. Thus, we genetically modified MSCs to overexpress CCR2 and investigated the targeted homing capacity and treatment efficacy of MSCCCR2 compared to those of the MSCvector control.Results: In vivo and ex vivo near-infrared fluorescence imaging showed that MSCCCR2 rapidly migrated and localized to injured livers in remarkably greater numbers following systemic infusion, and these cells were retained in liver lesions for a longer time than MSCvector. Furthermore, MSCCCR2 exhibited significantly enhanced efficacy in the treatment of ALF in mice, which was indicated by a dramatically improved survival rate, the alleviation of liver injury with reduced inflammatory infiltration, and hepatic apoptosis, and the promotion of liver regeneration.Conclusions: Altogether, these results indicate that CCR2 overexpression enhances the targeted migration of MSCs to damaged livers, improves their treatment effect, and may provide a novel strategy for improving the efficacy of cell therapy for ALF.

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Noncanonical Wnt5a/JNK Signaling Contributes to the Development of D-Gal/LPS-Induced Acute Liver Failure

10.21203/rs.3.rs-995106/v1 ◽

2021 ◽

Author(s):

Xiang-fen Ji ◽

Yu-chen Fan ◽

Fei Sun ◽

Jing-wei Wang ◽

kai wang

Keyword(s):

Liver Failure ◽

Acute Liver Failure ◽

Inflammatory Cytokines ◽

Dependent Manner ◽

Jnk Signaling ◽

Wnt5a Expression ◽

Jnk Activation

Abstract Acute liver failure (ALF) is a deadly clinical disorder with few effective treatments and unclear pathogenesis. In our previous study, we demonstrated that aberrant Wnt5a expression was involved in acute on chronic liver failure. However, the role of Wnt5a in ALF is unknown. We investigated the expression of Wnt5a and its downstream signaling of c-jun N-terminal kinase (JNK) in a mouse model of ALF established by co-injection of D-galactosamine (D-Gal) and lipopolysaccharide (LPS) in C57BL/6 mice. We also investigated the role of Box5, a Wnt5a antagonist in vivo. Moreover, the effect of Wnt5a/JNK signaling on downstream inflammatory cytokines expression, phagocytosis and migration in THP-1 macrophages was studied in vitro. Aberrant Wnt5a expression and JNK activation were detected in D-Gal/LPS-induced ALF mice. Box5 pretreatment reversed JNK activation, and eventually decreased the mortality rate of D-Gal/LPS-treated mice with reduced hepatic necrosis and apoptosis, serum ALT and AST levels, and liver inflammatory cytokines expression, although the last was not significant. We further demonstrated that recombined Wnt5a (rWnt5a) induced tumor necrosis α (TNF-α) and Interleukin-6 (IL-6) mRNA expression, and increased the phagocytosis ability of THP-1 macrophages in a JNK-dependent manner, which could be restored by Box5. In addition, rWnt5a-induced migration of THP-1 macrophages was also turned by Box5. Our findings suggested that Wnt5a/JNK signaling play important role in the development of ALF, and Box5 could have particular hepatoprotecive effects in ALF.

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In Vitro Effects of Sera From Children With Acute Liver Failure on Metabolic and Synthetic Activity of Cryopreserved Human Hepatocytes

Journal of Pediatric Gastroenterology and Nutrition ◽

10.1097/mpg.0b013e31819114da ◽

2009 ◽

Vol 48 (5) ◽

pp. 604-607 ◽

Cited By ~ 3

Author(s):

Ragai R Mitry ◽

Sanjay Bansal ◽

Robin D Hughes ◽

Giorgina Mieli-Vergani ◽

Anil Dhawan

Keyword(s):

Liver Failure ◽

Acute Liver Failure ◽

Human Hepatocytes ◽

Synthetic Activity ◽

In Vitro Effects

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Maintenance of rat hepatocytes under inflammation by coculture with human orbital fat-derived stem cells

Cellular & Molecular Biology Letters ◽

10.2478/s11658-012-0004-9 ◽

2012 ◽

Vol 17 (2) ◽

Cited By ~ 5

Author(s):

Xia Chen ◽

Shichang Zhang ◽

Tao Liu ◽

Yong Liu ◽

Yingjie Wang

Keyword(s):

Stem Cells ◽

Liver Failure ◽

Acute Liver Failure ◽

Functional Decline ◽

Rat Hepatocytes ◽

Rat Hepatocyte ◽

Orbital Fat ◽

Coculture System ◽

Marrow Mesenchymal Stem Cells

AbstractPreservation of hepatocyte functions in vitro will undoubtedly help the management of acute liver failure. The coculture system may be able to prevent functional decline of hepatocytes. It has already been shown that hepatocytes, when cocultured with bone marrow mesenchymal stem cells, could undergo long-term culture in vitro without loss of functions. In this study, human orbital fat-derived stem cells were isolated and cocultured with rat hepatocytes. When treated with serum from an acute liver failure patient, rat hepatocyte monoculture showed reduction of cell viability and loss of liverspecific functions. However, rat hepatocytes in the coculture system were still able to secret albumin and synthesize urea. IL-6 was significantly elevated in the coculture of rat hepatocyte with orbital fat-derived stem cells, and it might be the key immunoregulator which protects rat hepatocytes against inflammation. Our data confirmed that orbital fat-derived stem cells, or other adipose tissue-derived stem cells, are an ideal candidate to support rat hepatocyte functions in vitro.

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Human Fetal Hepatocyte Line, L-02, Exhibits Good Liver Function In Vitro and in an Acute Liver Failure Model

Transplantation Proceedings ◽

10.1016/j.transproceed.2012.09.121 ◽

2013 ◽

Vol 45 (2) ◽

pp. 695-700 ◽

Cited By ~ 25

Author(s):

X. Hu ◽

T. Yang ◽

C. Li ◽

L. Zhang ◽

M. Li ◽

...

Keyword(s):

Liver Function ◽

Liver Failure ◽

Acute Liver Failure ◽

Failure Model ◽

Good Liver Function ◽

Fetal Hepatocyte

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Three-dimensional bioprinted hepatorganoids prolong survival of mice with liver failure

Gut ◽

10.1136/gutjnl-2019-319960 ◽

2020 ◽

pp. gutjnl-2019-319960 ◽

Cited By ~ 2

Author(s):

Huayu Yang ◽

Lejia Sun ◽

Yuan Pang ◽

Dandan Hu ◽

Haifeng Xu ◽

...

Keyword(s):

Drug Metabolism ◽

Liver Failure ◽

Human Liver ◽

Three Dimensional ◽

3D Bioprinting ◽

Heparg Cells ◽

Liver Functions ◽

Human Specific

ObjectiveShortage of organ donors, a critical challenge for treatment of end-stage organ failure, has motivated the development of alternative strategies to generate organs in vitro. Here, we aim to describe the hepatorganoids, which is a liver tissue model generated by three-dimensional (3D) bioprinting of HepaRG cells and investigate its liver functions in vitro and in vivo.Design3D bioprinted hepatorganoids (3DP-HOs) were constructed using HepaRG cells and bioink, according to specific 3D printing procedures. Liver functions of 3DP-HOs were detected after 7 days of differentiation in vitro, which were later transplanted into Fah-deficient mice. The in vivo liver functions of 3DP-HOs were evaluated by survival time and liver damage of mice, human liver function markers and human-specific debrisoquine metabolite production.Results3DP-HOs broadly acquired liver functions, such as ALBUMIN secretion, drug metabolism and glycogen storage after 7 days of differentiation. After transplantation into abdominal cavity of Fah-/-Rag2-/- mouse model of liver injury, 3DP-HOs further matured and displayed increased synthesis of liver-specific proteins. Particularly, the mice acquired human-specific drug metabolism activities. Functional vascular systems were also formed in transplanted 3DP-HOs, further enhancing the material transport and liver functions of 3DP-HOs. Most importantly, transplantation of 3DP-HOs significantly improved the survival of mice.ConclusionsOur results demonstrated a comprehensive proof of principle, which indicated that 3DP-HO model of liver tissues possessed in vivo hepatic functions and alleviated liver failure after transplantation, suggesting that 3D bioprinting could be used to generate human liver tissues as the alternative transplantation donors for treatment of liver diseases.

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