scholarly journals Antiviral Effectivity of Favipiravir Against Peste Des Petits Ruminants Virus Is Mediated by the JAK/STAT and PI3K/AKT Pathways

2021 ◽  
Vol 8 ◽  
Author(s):  
Weifeng Zhang ◽  
Hualong Deng ◽  
Yanfen Liu ◽  
Shaohong Chen ◽  
You Liu ◽  
...  
Keyword(s):  
Western Blotting ◽  
Effective Means ◽  
Small Ruminants ◽  
Antiviral Agent ◽  
Vero Cells ◽  
Host Cells ◽  
Peste Des Petits Ruminants ◽  
Expression Levels ◽  
Qrt Pcr

Peste des petits ruminants virus (PPRV), belonging to the genus Morbillivirus in the family Paramyxoviridae, causes severe infectious disease in small ruminants and has been rapidly spreading in many parts of Africa, the Middle East, and Asia. Although vaccination is considered to be an effective means of controlling PPR, the heat-sensitive nature of the vaccines against PPRV greatly limits their application in areas with a hot climate. In the present study, we investigated the anti-PPRV effects of favipiravir and sought to identify the underlying mechanisms in vitro using the Vero cell line. MTT assays, Western blotting, indirect immunofluorescence assays, virus plaque formation assays, and qRT-PCR were used to assess the effects of favipiravir on the life cycle of PPRV and the expression of RNA-dependent RNA polymerase (RdRp). Additionally, the expression levels of JAK1, STAT1, phosphorylated (p)-STAT1, PI3K, AKT, and p-AKT, as well as those of signaling molecules acting downstream of the JAK/STAT and PI3K/AKT signaling pathways, were determined by Western blotting and qRT-PCR. The results indicated that, in PPRV-infected, favipiravir-treated Vero cells, the attachment, invasion, replication, and release of PPRV were significantly inhibited, as was the expression of RdRp, when compared with that in untreated PPRV-infected cells. Furthermore, in favipiravir-treated cells, the expression of JAK1 and STAT1 was downregulated, whereas that of p-STAT1 was significantly upregulated. Similarly, the expression levels of PKR, IRF9, ISG54, and MxA proteins that are associated with innate antiviral activity in host cells were also markedly increased. Moreover, with favipiravir treatment, the expression of PI3K and p-AKT and the p-AKT/AKT ratio were significantly decreased, whereas the expression of AKT was noticeably upregulated. The expression of GSK3, NF-κB p65, p-NF-κB p65, and BAD was also increased with favipiravir treatment, while the expression of CREB, p-CREB, p-GSK3, and Bcl-2 was slightly decreased. In addition, all the p-GSK3/GSK3, p-CREB/CREB, p-NF-κB/NF-κB, and p-BAD/BAD ratios were significantly reduced in favipiravir-treated cells. These results implied that the antiviral effectivity of favipiravir against PPRV is mediated by the JAK/STAT and PI3K/AKT pathways and that favipiravir has potential for use as an effective antiviral agent against PPRV.

Metformin Promotes the Hair-Inductive Activity of Three-Dimensional Aggregates of Epidermal and Dermal Cells Self-Assembled In Vitro

2021 ◽  
Author(s):  
Chao Sun ◽  
Shuang-Hai Hu ◽  
Bing-Qi Dong ◽  
Shan Jiang ◽  
Fang Miao ◽  
...  
Keyword(s):  
Hair Follicle ◽  
Western Blotting ◽  
Three Dimensional ◽  
Epidermal Cells ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Self Assembled ◽  
Dermal Cells ◽  
Inductive Activity

Introduction: Although it has been reported that the anti-diabetic drug metformin has multiple extra-hypoglycemic activities, such as anti-oxidation, anti-aging and even anti-tumor, topical metformin also can induce hair regeneration, but the precise mechanism involved in that process is still unclear. Objectives: To assess the effect of metformin on hair growth in a mouse hair follicle reconstitution model generated by in vitro self-assembled three-dimensional aggregates of epidermal and dermal cells (3D aggregates). Methods: Epidermal cells and dermal cells were isolated and cultured from the mouse skin of fifty C57BL/6 mouse pups (1-day-old). For tracing the distribution of dermal cells during the self-assembly process of 3D aggregates, the dermal cells were labeled with Vybrant Dil cell-labelling solution and mixed with epidermal cells at 1:1 ratio. Formed 3D aggregates were treated with 10 mM metformin and then were grafted into recipient BALB/c nude mice. The biomarkers (HGF, CD133, ALP, β-catenin and SOX2) associated with the hair-inductive activity of dermal cells were detected in the grafted skin tissues and in cultured 3D aggregates treated with metformin using immunofluorescent staining, quantitative real-time RT-PCR (qRT-PCR), and western blotting. Furthermore, the expression levels of CD133 were also examined in dermal cells with different passage numbers using qRT-PCR and western blotting. Results: Metformin directly stimulates the activity of alkaline phosphatase (ALP) of cultured 3D aggregates, upregulates both the protein and mRNA expression levels of molecular markers (HGF, CD133, ALP, β-catenin and SOX2) and improves the survival rate of reconstituted hair follicles. Moreover, we also found that metformin increases the expression of CD133 in dermal cells thus maintaining their trichogenic capacity that would normally be lost by serial subculture. Conclusions: These results suggest that metformin can promote hair follicle regeneration in vitro through up-regulation of the hair inductive capability of dermal cells, warranting further evaluation in the clinical treatment of male or female pattern hair loss.

Quantitative Analysis of Lysine Acetylation in Vero Cells Infected With Peste Des Petits Ruminants Virus

2021 ◽  
Author(s):  
Xuelian Meng ◽  
Xueliang Zhu ◽  
Rui Zhang ◽  
Zhidong Zhang
Keyword(s):  
Rna Virus ◽  
Small Ruminants ◽  
Vero Cells ◽  
Host Cells ◽  
Lysine Acetylation ◽  
Peste Des Petits Ruminants ◽  
Bioinformatics Analyses ◽  
Protein Protein Interaction ◽  
Quantification Analysis ◽  
And Function

Abstract Background: Peste des petits ruminants virus (PPRV) is a negative-stranded RNA virus belonging to the Paramyxoviridae family and causes acute, highly contagious disease in small ruminants. Lysine acetylation plays central role in regulating gene expression. However, the extent and function of lysine acetylation in host cells during PPRV infection remains unknown. Methods: Lysine acetylation of PPRV-infected Vero cells was tested and differentially expressed lysine acetylation was found. The acetylated peptides were enriched using specific antibody and labeled with demethylation. Proteins with acetylation sites were identified. Subsequently, intensive bioinformatics analysis of succinylome of PPRV-infected Vero cells was were performed. In this study, intensive proteomic quantification analysis of the proteome and acetylome of PPRV-infected Vero cells was performed using dimethylation labeling-based quantitative proteomics. Results: We identified 4729 cellular proteins and 1068 proteins with 2641 modification sites quantifiable detected by mass spectrometry, of which 304 proteins with 410 acetylation sites were significantly acetylated in response to PPRV infection. Bioinformatics analyses revealed that the differentially acetylated proteins mainly participated in carbohydrate catabolic and DNA metabolic process, and were associated with multifarious functions, suggesting that intracellular activities were extensively changed after PPRV infection. Protein-protein interaction (PPI) network of the identified proteins further indicated that a variety of chaperone and ribosome processes were modulated by acetylation. Conclusions: To our knowledge, this is the first study on acetylome in host cell infected with PPRV. It provides an important baseline to future study the roles of acetylation in the host response to PPRV replication.

Abstract 1772: Cardiomyocyte Damage-Released S100A1 Protein Evokes A Proinflammatory Phenotype In Cardiac Fibroblasts

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
David Rohde ◽  
Melanie Boerries ◽  
Herzog Nicole ◽  
Gang Qiu ◽  
Philipp Ehlermann ◽  
...  
Keyword(s):  
Western Blotting ◽  
Immune Cells ◽  
Nuclear Translocation ◽  
Cardiac Fibroblasts ◽  
Innate Immune ◽  
Host Cells ◽  
Boyden Chamber ◽  
Innate Immune Cells ◽  
Danger Signal

Background: S100A1, a cardiomyocyte specific inotropic calcium sensor protein, is released from infarcted human myocardium in the extracellular environment and circulation, reaching peak serum levels (1–2 μM) 8–9 hours after clinical onset. As growing evidence indicates that S100 proteins can act as pre-existing danger signals triggering the innate immune system into action upon release from injured host cells, we hypothesized that damage-released S100A1 can act as a cardiac danger signal alerting innate immune cells. Methods and Results: Here we report for the first time that necrotic cardiomyocytes release S100A1 protein in vitro, which is exclusively internalized by cardiac fibroblasts (CFs) in a clathrin- and caveolin-independent manner as shown by IF. Internalized S100A1 specifically activated MAPKs/SAPKs (p38, ERK1/2 and JNK) resulting in nuclear translocation of p65 (NF-kB) as assessed by Western blotting, EMSA and IF. In turn, S100A1 triggered an inflammatory gene program in CFs including enhanced expression of adhesion molecules, integrins, chemokines and cytokines including I-CAM, V-CAM, CD11b/18, IL1-alpha, MCP-1, TNF-alpha, SDF-1 among others as obtained by RT-PCR, Western blotting and ELISA. This resulted in enhanced chemoattraction and adhesion of monocytotic and stem cells to S100A1-activated CF as shown by Boyden-chamber and adhesion assays. In line with their proinflammatory transition, S100A1-activated CFs exhibited decreased collagen-1/-3 expression and de-novo collagen production, enhanced collagenolytic MMP-9 abundance and activity and increased levels of the antiangiogenic matricellular factor thrombospondin-2 reflecting extracellular matrix net degradation. Importantly, the immun-modulatory and antifibrotic actions of S100A1 protein in vitro were restricted to CFs, RAGE independent and occurred at concentrations (0.1–1 μM) that were found in patients after AMI. Conclusion: Our in vitro results indicate that S100A1 has the properties of a pre-exisiting endogenous cardiomyocyte danger signal transforming cardiac fibroblasts into immunmodulatory cells that might recruit innate immune cells to the site of cardiac injury and link cardiomyocyte damage to post-MI inflammation.

scholarly journals Understanding Pathogen-host Interplay by Expression Profiles of LncRNA and mRNA in the Liver of Echinococcus Multilocularis-infected Mice

2021 ◽  
Author(s):  
Xiaofeng Nian ◽  
Li Li ◽  
Xusheng Ma ◽  
Xiurong Li ◽  
Wenhui Li ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Potential Function ◽  
Western Blotting ◽  
Echinococcus Multilocularis ◽  
Th17 Cell ◽  
Antigen Processing ◽  
Expression Profiles ◽  
Expression Levels ◽  
Amino Terminal ◽  
Qrt Pcr

Abstract Background: Echinococcus multilocularis (Em) infection and the growth and proliferation of its metacestode within the internal organs of hosts are related to complex host–parasite interactions at the molecular level. Previous studies reported the profiles of long non-coding RNAs (lncRNAs) and mRNAs in Echinococcus granulosus-infected mice or cells, suggesting the potential role of lncRNAs in regulating host-parasite interplay. However, the profiles of lncRNAs and mRNAs of mice in response to Em are poorly understood. Methods: Numerous differentially expressed lncRNAs (DELs) and mRNAs (DEMs) in the mouse liver at eight time points after Em infection were identified by microarray. Functional Annotation of dysregulated DEMs was conducted by gene ontology (GO) classification and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The potential function of DELs was predicted by constructing lncRNA-mRNA co-expression network and Transcription factor (TF)-lncRNA-mRNA Ternary Network. Additionally, qRT-PCR and western blotting were used to validate the upregulated DEMs at 30 days post-infection (dpi), which were enriched in Toll-like and RIG-I-like receptor signaling pathways. Cytokines and chemokines involved in these two pathways were determined by ELISA.Results: Thirty-one DEMs and 68 DELs were found continuously dysregulated. These DEMs were notably enriched in the “antigen processing and presentation,” “Th1 and Th2 cell differentiation” and “Th17 cell differentiation” pathways. The potential function prediction of DELs revealed that most DELs might influence the differentiation of Th17 cell and TGF-β/Smad pathway through trans regulating the SMAD3, STAT1, and early growth response (EGR) genes. Additionally, the validated results by qRT-PCR and western blotting showed that the mRNA expression levels of these genes increased while the corresponding protein expression levels were unaltered except c-Jun amino-terminal kinase (JNK). Regardless, phospho-nuclear factor Kappa B (p-NF-κB) downstream of these two pathways was induced at 15 and 30 dpi, which led to the elevated levels of IL-1 beta and IL-6 in the serum. Conclusion: Our data provide novel clues in understanding the roles of lncRNAs in the host–Em interplay and the influence of Em infection on host innate immunity.

scholarly journals Oxymatrine Ameliorates Memory Impairment in Diabetic Rats by Regulating Oxidative Stress and Apoptosis: Involvement of NOX2/NOX4

2020 ◽  
Vol 2020 ◽  
pp. 1-15
Author(s):  
Yongpan Huang ◽  
Xinliang Li ◽  
Xi Zhang ◽  
Jiayu Tang
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Brain Injury ◽  
Western Blotting ◽  
Caspase 3 ◽  
Memory Function ◽  
Diabetic Rats ◽  
Oxygen Species ◽  
Expression Levels

Oxymatrine (OMT) is the major quinolizidine alkaloid extracted from the root of Sophora flavescens Ait and has been shown to exhibit a diverse range of pharmacological properties. The aim of the present study was to investigate the role of OMT in diabetic brain injury in vivo and in vitro. Diabetic rats were induced by intraperitoneal injection of a single dose of 65 mg/kg streptozotocin (STZ) and fed a high-fat and high-cholesterol diet. Memory function was assessed using a Morris water maze test. A SH-SY5Y cell injury model was induced by incubation with glucose (30 mM/l) to simulate damage in vitro. The serum fasting blood glucose, insulin, serum S100B, malondialdehyde (MDA), and superoxide dismutase (SOD) levels were analyzed using commercial kits. Morphological changes were observed using Nissl staining and electron microscopy. Cell apoptosis was assessed using Hoechst staining and TUNEL staining. NADPH oxidase (NOX) and caspase-3 activities were determined. The effects of NOX2 and NOX4 knockdown were assessed using small interfering RNA. The expression levels of NOX1, NOX2, and NOX4 were detected using reverse transcription-quantitative PCR and western blotting, and the levels of caspase-3 were detected using western blotting. The diabetic rats exhibited significantly increased plasma glucose, insulin, reactive oxygen species (ROS), S-100B, and MDA levels and decreased SOD levels. Memory function was determined by assessing the percentage of time spent in the target quadrant, the number of times the platform was crossed, escape latency, and mean path length and was found to be significantly reduced in the diabetic rats. Hyperglycemia resulted in notable brain injury, including histological changes and apoptosis in the cortex and hippocampus. The expression levels of NOX2 and NOX4 were significantly upregulated at the protein and mRNA levels, and NOX1 expression was not altered in the diabetic rats. NOX and caspase-3 activities were increased, and caspase-3 expression was upregulated in the brain tissue of diabetic rats. OMT treatment dose-dependently reversed behavioral, biochemical, and molecular changes in the diabetic rats. In vitro, high glucose resulted in increases in reactive oxygen species (ROS), MDA levels, apoptosis, and the expressions of NOX2, NOX4, and caspase-3. siRNA-mediated knockdown of NOX2 and NOX4 decreased NOX2 and NOX4 expression levels, respectively, and reduced ROS levels and apoptosis. The results of the present study suggest that OMT alleviates diabetes-associated cognitive decline, oxidative stress, and apoptosis via NOX2 and NOX4 inhibition.

Clonal and Subclonal Cytogenetic Aberrations: Association with D-Type Cyclin Expression and Event-Free Survival (EFS) in Multiple Myeloma (MM).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3439-3439
Author(s):  
Dirk Hose ◽  
John DeVos ◽  
Nadine Müller ◽  
Jean-Francois Rossi ◽  
Christiane Heiß ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Western Blotting ◽  
Plasma Cells ◽  
Early Event ◽  
Expression Data ◽  
Rt Pcr ◽  
Qrt Pcr ◽  
Cytogenetic Aberrations ◽  
Significant Difference

Abstract AIM. Expression changes of D-type cyclins are thought to be an early event in the genesis of Multiple Myeloma and are associated with distinct cytogenetic aberrations. These aberrations appear with different percentages (“clonal” or “subclonal”) in a given patient. We assessed whether the height of CCND expression assessed by gene expression profiling and quantitative RT-PCR (qRT-PCR) correlates with the presence of clonal or subclonal aberrations of 11q13, t(11;14) and t(4;14). PATIENTS AND METHODS. 128 newly diagnosed MM-patients (65 training (TG)/63 independent validation group (VG)) and 14 normal donors (ND) were included. Bone marrow aspirates were CD138-purified by activated magnetic cell sorting. RNA was in-vitro transcribed and hybridised to Affymetrix HG U133 A+B GeneChip (TG) and HG U133 2.0 plus array (VG). CCND1 and CCND2 expression was verified by real time RT-PCR and western blotting. iFISH was performed on purified MM-cells using probesets for chromosomes 1q21, 9q34, 11q23, 11q13, 13q14, 15q22, 17p13, 19q13, 22q11 and the translocations t(4;14) and t(11;14). Clonal aberrations were defined as being present in >60%, subclonal aberrations in 20 to 60% of MMC in a given patient. Expression data were gcrma normalised and a Kruskal-Wallis rank sum test used (Bioconductor). RESULTS. 11q13+. CCND1 (208711_s_at, 208712_at) is significantly higher (p<0.0001), CCND2 (200953_s_at, 200951_s_at) significantly lower (p<0.0001) expressed in MMC harbouring clonal, compared to subclonal, or no gain of 11q13. t(11;14). CCND1 is significantly higher (p<0.0001), CCND2 significantly lower (p<0.0001) expressed in MMC harbouring clonal, compared to subclonal, or no t(11;14). t(4;14). CCND1 is significantly lower (p<0.0001), CCND2 significantly higher (p<0.0001) expressed in MMC harbouring clonal compared to subclonal, or no t(4;14). The expression of CCND3 (201700_at) is not significantly different between the 3 groups for all aberrations investigated. CCND2 and CCND3, but not CCND1 are expressed by normal plasma cells. Results have been verified by qRT-PCR (n=40) for CCND1 and CCND2. Expression of CCND1, CCND2 and CCND3 has been verified by western blotting on selected samples. The expression of CCND2 correlates with short EFS, but not if patients with t(4;14) are excluded. There is no significant difference in EFS for patients harbouring the respective aberrations in a clonal or subclonal pattern. CONCLUSION. An additional copy of 11q13 or t(11;14) correlates with increased CCND1 and decreased CCND2 expression, a t(4;14) is associated with an increase of CCND2 and a decrease of CCND1 expression. In each case, the height of the CCND-expression is significantly different whether the respective aberration is clonal or subclonal. Thus, when interpreting expression data in the context of cytogenetic aberrations, it is important to consider if plasma cells carry a respective aberration in a subclonal/clonal pattern.

Molecular cloning, characterization, and immune response against white spot syndrome virus and Taura syndrome virus infections of peroxiredoxin in Litopenaeus vannamei and its antioxidant activity

Crustaceana ◽  
2015 ◽  
Vol 88 (10-11) ◽  
pp. 1149-1161 ◽  
Author(s):  
Digang Zeng ◽  
Min Peng ◽  
Xiuli Chen ◽  
Chunling Yang ◽  
Xiaohan Chen ◽  
...  
Keyword(s):  
Immune Response ◽  
Litopenaeus Vannamei ◽  
White Spot Syndrome Virus ◽  
Phylogenetic Analyses ◽  
Virus Infections ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Oxidative Stresses ◽  
Different Doses

Peroxiredoxin (Prx) is an important peroxidase that can protect organisms against various oxidative stresses. In this study, a member of Prx family, designated LvPrx, was cloned fromLitopenaeus vannamei. Sequence and phylogenetic analyses indicated that LvPrx belongs to the 2-Cys Prx (Prx IV) isoform. The recombinant LvPrx protein was constructed and expressed inEscherichia coli, and the purified LvPrx proteins were shown to reduce H2O2in vitro in the presence of dithiothreitol, indicating that LvPrx is a functional peroxiredoxin. Using qRT-PCR, the mRNA expression levels of LvPrx were determined in the haemocytes ofL. vannameiat different stages after being challenged with WSSV and TSV at different doses. The results showed that the expression levels of LvPrx were significantly up-regulated () during 4-24 h after both WSSV and TSV challenge, suggesting that LvPrx may participate in the shrimp’s immune response to viral infection.

scholarly journals Attachment and invasion ofToxoplasma gondiiandNeospora caninumto epithelial and fibroblast cell linesin vitro

Parasitology ◽  
2005 ◽  
Vol 131 (5) ◽  
pp. 583-590 ◽  
Author(s):  
YING LEI ◽  
M. DAVEY ◽  
J. T. ELLIS
Keyword(s):  
Cell Lines ◽  
Neospora Caninum ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Vero Cells ◽  
Host Cells ◽  
Trypsin Treatment ◽  
Epithelial Cell Lines ◽  
Pre Treatment

Attachment and invasion ofToxoplasma gondiiandNeospora caninumto a cat and a dog fibroblast cell line and 2 epithelial cell lines (a cat kidney and Vero) were comparedin vitrousing fluorescence antibody methodology. In addition, trypsin treatment of tachyzoites was used to determine whether protein molecules were essential to the process of invasion. The results show that bothT. gondiiandN. caninuminvaded all 4 cell lines, and that pre-treatment ofT. gondiitachyzoites with trypsin caused an increase in the ability of the parasite to invade these host cells. FurthermoreT. gondii, in comparison toN. caninum, invaded all 4 cell lines at greater levels. The results here support the conclusion that bothT. gondiiandN. caninumhave the ability to invade a variety of cell types including both dog and cat cells, and questions the utility of Vero cells as an appropriate host cell forin vitrostudies on the biology of these taxa.

scholarly journals A Polyphenol Rich Extract from Solanum melongena L. DR2 Peel Exhibits Antioxidant Properties and Anti-Herpes Simplex virus Type 1 Activity In Vitro

2018 ◽  
Author(s):  
Antonella Di Sotto ◽  
Silvia Di Giacomo ◽  
Donatella Amatore ◽  
Marcello Locatelli ◽  
Annabella Vitalone ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Antioxidant Properties ◽  
Solanum Melongena ◽  
Vero Cells ◽  
Host Cells ◽  
Reducing Conditions ◽  
Simplex Virus ◽  
Hsv 1

DR2B and DR2C extracts, from peel of commercially and physiologically ripe eggplants, were studied for the antioxidative cytoprotective properties and anti-HSV-1 activity, in line with the evidence that several antioxidants can impair viral replication by maintaining reducing conditions into the host cells. The antioxidative cytoprotective effects against tBOOH-induced damage was assessed in Caco2 cells, while the antiviral activity was studied in Vero cells; phenolic and anthocyanin fingerprint was characterized by integrated phytochemical methods. Results highlighted different compositions of the extracts, with chlorogenic acid and delphinidin-3-rutinoside as the major constituents; other peculiar phytochemicals were also identified. DR2C resulted able to partly counteract the tBOOH-induced cytotoxicity, with a remarkable lowering of lactate metabolism under both normoxia and hypoxia. DR2B and DR2C reduced ROS production, possessed scavenging and chelating properties. Interestingly, DR2C increased intracellular GSH levels. Furthermore, DR2C inhibited the HSV-1 replication when added for 24 h after viral adsorption, as also confirmed by the reduction of many viral proteins expression. Since DR2C was able to reduce NOX4 expression during HSV-1 infection, its antiviral activity may be correlated to its antioxidant properties. Although further studies are needed to better characterize DR2C activity, the results suggest this extract as a promising new anti-HSV-1 agent.

scholarly journals The Decrease in Farnesoid X Receptor, Pregnane X Receptor and Constitutive Androstane Receptor in the Liver after Intestinal Ischemia-Reperfusion

2012 ◽  
Vol 15 (5) ◽  
pp. 616 ◽  
Author(s):  
Jiro Ogura ◽  
Yusuke Terada ◽  
Takashi Tsujimoto ◽  
Takahiro Koizumi ◽  
Kaori Kuwayama ◽  
...  
Keyword(s):  
Western Blotting ◽  
Hepg2 Cells ◽  
Intestinal Ischemia ◽  
Ischemia Reperfusion ◽  
Constitutive Androstane Receptor ◽  
Pregnane X Receptor ◽  
Farnesoid X Receptor ◽  
Expression Levels ◽  
Intestinal Ischemia Reperfusion

Purpose. Intestinal ischemia-reperfusion (I/R) damages remote organs, including the liver, and promotes multi-organ failure (MOF). However, the molecular mechanisms underlying acute liver injury after intestinal I/R have not been completely elucidated. Farnesoid X receptor (FXR), pregnane X receptor (PXR) and constitutive androstane receptor (CAR) regulate metabolizing enzymes and transporters, and coordinately prevent hepatotoxicity reflecting an inability of appropriate excretion of endogenous toxic compounds. In this study, we assessed FXR, PXR and CAR expression levels and their localization levels in nuclei in the liver after intestinal I/R. We also investigated the effect of IL-6 on FXR, PXR and CAR expression levels and their localization levels in nuclei in in vitro experiments. Methods. We used intestinal I/R model rats. Moreover, HepG2 cells were used in in vitro study. Real-time PCR and Western blotting were used to assess mRNA and protein expression levels. Nuclear receptor localization in nuclei was analyzed by Western blotting using nuclear extracts. Results. FXR and PXR expression levels began to be decreased at 3 h, and FXR, PXR and CAR expression levels were decreased at 6 h after intestinal I/R. The localization levels of FXR, PXR and CAR in nuclei began to be decreased at 3 h, and all of them were decreased at 6 h after intestinal I/R. In HepG2 cells, FXR, PXR and CAR expression levels were decreased by 0.5-1 ng/mL, 0.5-100 ng/mL and 100 ng/mL IL-6 treatment for 24 h, respectively. FXR, PXR and CAR localization levels in nuclei were suppressed by 0.5-10 ng/mL, 10-100 ng/mL and 10-100 ng/mL IL-6 treatment for 24 h, respectively. Conclusions. FXR, PXR and CAR expression levels are decreased in the liver after intestinal I/R. IL-6 is one of main causes the decreases in expressions of these receptors. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

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