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Hepatocyte Isolation

Qin Wang, Yue Yin, Weizhen Zhang
Obese mice demonstrate disruption of the circadian clock and feeding cycle. Circulating ghrelin, a hormone secreted mainly by gastric X/Alike cells, is significantly reduced in obese humans and animals. Here, we examined whether ghrelin improves the disruption of the circadian rhythm in steatotic hepatocytes and liver. The effects of ghrelin on hepatic circadian clock genes were studied in steatotic hepatocytes and liver of mice fed a high-fat diet (HFD) for 12 weeks. The circadian clock of cultured hepatocytes was synchronized by treatment with 100 nM dexamethasone for 1 h...
October 12, 2018: International Journal of Molecular Sciences
Octavio Galindo-Hernández, Iván Córdova-Guerrero, Laura Janeth Díaz-Rubio, Ángel Pulido-Capiz, José Fernando Díaz-Villanueva, César Yahel Castañeda-Sánchez, Nicolás Serafín-Higuera, Víctor García-González
Endoplasmic reticulum stress is a cellular phenomenon that has been associated with metabolic disorders, contributing to the development of obesity, fatty liver disease, and dyslipidemias. Under metabolic overload conditions, in cells with a high protein-secretory activity, such as hepatocytes and Langerhans β cells, the unfolded protein response (UPR) is critical in to maintain protein homeostasis (proteostasis). UPR integrated by a tripartite signaling system, through activating transcription factor 6, protein kinase R-like endoplasmic reticulum kinase (PERK), and inositol-requiring enzyme 1, regulates gene transcription and translation to resolve stress and conserve proteostasis...
October 15, 2018: Journal of Cellular Biochemistry
Yasen Aimaiti, Xin Jin, Yue Shao, Wei Wang, Dewei Li
Hepatic stellate cells (HSCs) play an important microenvironmental role in hepatic progenitor cells (HPCs) differentiation fate. To reveal the specific mechanism of HSCs induced by transforming growth factor β1 (TGF-β1) signaling in HPCs differentiation process, we used Knockin and knockdown technologies induced by lentivirus to upregulate or downregulate TGF-β1 level in mouse HSCs (mHSCs) (mHSCs-TGF-β1 or mHSCs-TGF-βR1sih3). Primary mouse HPCs (mHPCs) were isolated and were cocultured with mHSCs-TGF-β1 and mHSCs-TGF-βR1sih3 for 7 days...
October 14, 2018: Journal of Cellular Physiology
Y F Zhou, Z Zhou, F Batistel, I Martinez-Cortés, R T Pate, D L Luchini, J J Loor
Insufficient supply of Met and choline (Chol) around parturition could compromise hepatic metabolism and milk protein synthesis in dairy cows. Mechanistic responses associated with supply of Met or Chol in primary liver cells enriched with hepatocytes (PHEP) from cows have not been thoroughly ascertained. Objectives were to isolate and culture PHEP to examine abundance of genes and proteins related to transmethylation, transsulfuration, and cytidine 5'-diphosphocholine (CDP-choline) pathways in response to Met or Chol...
October 10, 2018: Journal of Dairy Science
Suyu Gao, Peng Zhang, Cuiyun Zhang, Fengyan Bao, Hua Li, Lixia Chen
Four meroterpenoids, applanatumols F (1), H (3), I (2), and lingzhiol (4) were isolated from the 95% EtOH extract of the fruiting bodies of Ganoderma sinense. Their structures were established on the basis of NMR spectroscopic analyses, optical rotatory dispersion data, ECD spectra, and X-ray crystallography. Compounds 1, 2, 4 existed as racemic mixtures ((+) 1a, 2a, 4a; (-) 1b, 2b, 4b), while 3 as a single enantiomer. Base on the seperated enantiomers, we sought to explicit possible effects of compounds 1-4 on hydrogen peroxide (H2 O2 )-induced cell death and to determine their underlying molecular mechanisms in human normal liver LO2 cells...
October 8, 2018: Fitoterapia
Wanfang Zhu, Shengzhi Su, Yunhui Xu, Zijian Xie, Yidan Bai, Wenyuan Liu, Masahiko Abe, Toshihiro Akihisa, Feng Feng, Jie Zhang
Three new C21 steroids, i.e., (3β,17α,20S)-pregn-5(6)-ene-3, 17, 20-triol-3-O-β-D- digitalopyranosyl-(1→4)-β-D-digitalopyranoside (4), (3β,17α,20S)- pregn-5(6)-ene-3, 17, 20-triol-20-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl-(1→2)-β-D-digital- opyranoside (8), (3β, 20R)-pregn-14(15)-ene-3, 20, 21-triol-3-O-β-D-glucopy- ranoside (10), along with ten known C21 steroids were isolated from Streptocaulon juventas. Their structures were elucidated on the basis of 1D and 2D NMR spectroscopic techniques, mass spectrometry as well as comparison with the literature...
October 5, 2018: Steroids
Jose Cordoba-Chacon, Andre Sarmento-Cabral, Mercedes Del Rio-Moreno, Alberto Diaz-Ruiz, Papasani V Subbaiah, Rhonda D Kineman
Nonalcoholic fatty liver disease (NAFLD), which includes non-alcoholic steatohepatitis (NASH), is associated with reduced growth hormone (GH) input/signaling and GH therapy is effective in the reduction/resolution of NAFLD/NASH in select patient populations. Our laboratory has focused on isolating the direct vs. indirect effects of GH in preventing NAFLD/NASH. We reported that chow-fed, adult-onset hepatocyte-specific GH receptor knockdown (aHepGHRkd) mice rapidly (within 7d) develop steatosis associated with increased hepatic de novo lipogenesis (DNL), independent of changes in systemic metabolic function...
October 5, 2018: Endocrinology
Farhana Afroz, Alwyn Kist, Jin Hua, Yabin Zhou, Elke M Sokoya, Robert Padbury, Vincent Nieuwenhuijs, Greg Barritt
Rapamycin (sirolimus) is employed as an immunosuppressant following liver transplant, to inhibit the re-growth of cancer cells following liver resection for hepatocellular carcinoma (HCC), and for the treatment of advanced HCC. Rapamycin also induces the expression of antioxidant enzymes in the liver, suggesting that pretreatment with the drug could provide a potential strategy to reduce ischemia reperfusion injury following liver surgery. The aim of this study was to further investigate the actions of rapamycin in inducing expression of the antioxidant enzymes heme oxygenase-1 (HO-1) and peroxiredoxin-1 (Prx-1) in normal liver and in tumorigenic liver cells...
October 2, 2018: Experimental and Molecular Pathology
Brian C Geohagen, Boris Korsharskyy, Amaresh Vydyanatha, Lars Nordstroem, Richard M LoPachin
Phloretin (Phl) is a dihydrochalcone flavonoid with significant cytoprotective properties; e.g., free radical trapping, electrophile scavenging. Based on this, it has been suggested that Phl might be useful in the treatment of pathogenic processes and prevention of drug toxicities. Therefore, we determined the ability of Phl to provide route- and dose-dependent hepatoprotection in a mouse model of acetaminophen (APAP) overdose. Intraperitoneal (i.p.) administration of Phl produced a bimodal effect; i.e., the highest dose (2...
October 1, 2018: Chemico-biological Interactions
Nicholas John Hunt, Glen Lockwood, Alessandra Warren, Hong Mao, Peter McCourt, David G Le Couteur, Victoria Carroll Cogger
Fenestrations are pores within liver sinusoidal endothelial cells (LSECs) that enable the transfer of substrates (particularly insulin and lipoproteins) between blood and hepatocytes. With increasing age there are marked reductions in the frequency and diameter of fenestrations, referred to as pseudocapillarization. Currently, fenestrations are thought to be regulated by VEGF and NO pathways promoting remodelling of the actin cytoskeleton and cell membrane lipid rafts. We investigated the effects of drugs that act on these pathways on fenestrations in old (18-24m) and young mice (3-4m)...
October 4, 2018: American Journal of Physiology. Gastrointestinal and Liver Physiology
Honghao Zheng, Wang Du, Yiyuan Duan, Keyu Geng, Jun Deng, Changyou Gao
The biomimetic anisotropic particles have different physicochemical properties on the opposite two sides, enabling diverse applications in emulsion, photonic display, and diagnosis. However, the traditional anisotropic particles have a very small size, ranging from sub-microns to a few microns. The design and fabrication of anisotropic macron-sized particles with new structures and properties is still challenging. In this study, anisotropic polycaprolactone (PCL) microparticles well separated with each other were prepared by crystallization from the dilute PCL solution in a porous 3D gelatin template...
October 4, 2018: ACS Applied Materials & Interfaces
Ying-Yin Sun, Yu-Xin Zhao, Xiao-Feng Li, Cheng Huang, Xiao-Ming Meng, Jun Li
Alcoholic liver disease (ALD) is a complex process that includes a wide range of hepatic lesions, from steatosis to cirrhosis, and even hepatocellular carcinoma (HCC). Accumulating evidence shows that the cytotoxic effects of ethanol metabolism lead to cell apoptosis and necrosis in ALD. Recently, several studies revealed that multifunctional protein β-arrestin 2 (Arrb2) modulated cell apoptosis in liver fibrosis and HCC, but its role in ALD has not been fully understood. The aim of this study is to explore the function and underlying mechanism of Arrb2 in hepatocyte survival and apoptosis in ALD...
2018: Frontiers in Pharmacology
Romina Fiorotto, Mariangela Amenduni, Valeria Mariotti, Luca Fabris, Carlo Spirli, Mario Strazzabosco
Liver diseases negatively impact the quality of life and survival of patients, and often require liver transplantation in cases that progress to organ failure. Understanding the cellular and molecular mechanisms of liver development and pathogenesis has been a challenging task, in part for the lack of adequate cellular models directly relevant to the human diseases. Recent technological advances in the stem cell field have shown the potentiality of induced pluripotent stem cells (iPSC) and liver organoids as the next generation tool to model in vitro liver diseases...
September 5, 2018: Biochimica et biophysica acta. Molecular basis of disease
Nina S Liland, Karin Pittman, Paul Whatmore, Bente E Torstensen, Nini H Sissener
Several feeding trials with Atlantic salmon fed naturally high phytosterol concentrations due to dietary rapeseed oil inclusion have shown changes in lipid metabolism and increased hepatic lipid storage in the fish. An in vitro trial with Atlantic salmon hepatocytes was, therefore, performed to study the possible direct effects of phytosterols on lipid storage and metabolism. The isolated hepatocytes were exposed to seven different sterol treatments and gene expression, as well as lipid accumulation by Oil Red O dyeing, was assessed...
July 2018: Lipids
Sander Lefere, Frederique Van de Velde, Anne Hoorens, Sarah Raevens, Sanne Van Campenhout, Astrid Vandierendonck, Sara Neyt, Bert Vandeghinste, Christian Vanhove, Charlotte Debbaut, Xavier Verhelst, Jo van Dorpe, Christophe Van Steenkiste, Christophe Casteleyn, Bruno Lapauw, Hans Van Vlierberghe, Anja Geerts, Lindsey Devisscher
Angiogenesis contributes to the development of non-alcoholic steatohepatitis (NASH) and promotes inflammation, fibrosis and progression to hepatocellular carcinoma (HCC). Angiopoietin-2 (Ang-2) is a key regulator of angiogenesis. We aimed to investigate the role of Ang-2 and its potential as therapeutic target in NASH using human samples, in vivo mouse models and in vitro assays. Serum Ang-2 levels were determined in 104 obese patients undergoing bariatric surgery and concomitant liver biopsy. The effect of the Ang-2/Tie2 receptor inhibiting peptibody L1-10 was evaluated in the methionine-choline deficient (MCD) and streptozotocin-western diet NAFLD mouse models, and in vitro on endothelial cells and bone marrow-derived macrophages...
September 26, 2018: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
Hongxian Yan, Qian Wang, Quan Shen, Zhaohui Li, Jianguo Tian, Qingfeng Jiang, Linbo Gao
Aim: This study aimed to investigate the key transcription factors (TFs), long noncoding RNAs (lncRNAs), and microRNAs (miRNAs) associated with hepatocellular carcinoma (HCC). Materials and Methods: The datasets GSE31383 and GSE54238 were downloaded from Gene Expression Omnibus data repository. GSE31383 was used to screen differentially expressed miRNAs, and GSE54238 was used to screen differentially expressed messenger RNAs (mRNAs) and lncRNAs. ChipBase was used to identify TF-miRNA pairs...
September 2018: Journal of Cancer Research and Therapeutics
Giulio Giustarini, Nienke Vrisekoop, Laura Kruijssen, Laura Wagenaar, Selma van Staveren, Manon van Roest, Rob Bleumink, Marianne Bol-Schoenmakers, Richard Weaver, Leo Koenderman, Joost Smit, Raymond Pieters
The fluoroquinolone trovafloxacin (TVX) is associated with a high risk of drug-induced liver injury (DILI). Although part of the liver damage by TVX+TNF relies on neutrophils, we have recently demonstrated that liver recruitment of monocytes and neutrophils is delayed by TVX.Here we show that the delayed leukocyte recruitment is caused by a combination of effects which are linked to the capacity of TVX to block the hemichannel pannexin 1. TVX inhibited find-me signal release in apoptotic HepG2 hepatocytes, decelerated freshly isolated human neutrophils toward IL-8 and f-MLF, and decreased the liver expression of ICAM-1...
September 22, 2018: Toxicological Sciences: An Official Journal of the Society of Toxicology
Yohan Kim, Kyojin Kang, Seung Bum Lee, Daekwan Seo, Sangtae Yoon, Heung Mo Yang, Sung Joo Kim, Kiseok Jang, Yun Kyung Jung, Kyeong Geun Lee, Valentina M Factor, Jaemin Jeong, Dongho Choi
BACKGROUND & AIMS: Currently, much effort is directed towards the development of new cell sources for clinical therapy using cell fate conversion approaches by small molecules. Direct lineage reprogramming to a progenitor state has been reported in terminally differentiated rodent hepatocytes, yet remains a challenge in human hepatocytes. METHODS: Human hepatocytes were isolated from healthy and diseased donor livers and reprogrammed into progenitor cells by two small molecules, A83-01 and CHIR99021 (AC), in the presence of EGF and HGF...
September 18, 2018: Journal of Hepatology
Chian-Jiun Liou, Ciao-Han Wei, Ya-Ling Chen, Ching-Yi Cheng, Chia-Ling Wang, Wen-Chung Huang
BACKGROUND/AIMS: Fisetin is a naturally abundant flavonoid isolated from various fruits and vegetables that was recently identified to have potential biological functions in improving allergic airway inflammation, as well as anti-oxidative and anti-tumor properties. Fisetin has also been demonstrated to have anti-obesity properties in mice. However, the effect of fisetin on nonalcoholic fatty liver disease (NAFLD) is still elusive. Thus, the present study evaluated whether fisetin improves hepatic steatosis in high-fat diet (HFD)-induced obese mice and regulates lipid metabolism of FL83B hepatocytes in vitro...
2018: Cellular Physiology and Biochemistry
Britt A Blokker, Monica Maijo, Marta Echeandia, Mikel Galduroz, Angela M Patterson, Anna Ten, Mark Philo, Rebecca Schungel, Virginia Gutierrez-de Juan, Emina Halilbasic, Claudia Fuchs, Gwenaelle Le Gall, Malgorzata Milkiewicz, Piotr Milkiewicz, Jesus M Banales, Simon M Rushbrook, José M Mato, Michael Trauner, Michael Müller, María Luz Martínez-Chantar, Marta Varela-Rey, Naiara Beraza
Cholestasis comprises aetiologically heterogeneous conditions characterized by accumulation of bile acids in the liver that actively contribute to liver damage. Sirtuin 1 (SIRT1) regulates liver regeneration and bile acid metabolism via modulating the farnesoid X receptor (FXR); we here investigate its role in cholestatic liver disease. We determined SIRT1 expression in livers from patients with cholestatic disease, in two experimental models of cholestasis, as well as in human and murine liver cells in response to bile acid loading...
September 19, 2018: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
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