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Mast cells (MCs) induce ductular reaction mimicking liver injury in mice via MC-derived TGF-β1 signaling

Mast cells (MCs) induce ductular reaction mimicking liver injury in mice via MC-derived TGF-β1 signaling

Background: Following liver injury, mast cells (MC) migrate to the liver and is activated in patients with cholestasis. Inhibition of MC decreased mediator ductular reaction (DR) and liver fibrosis. TGF-β1 contributes to fibrosis and promote liver disease.


Objective: To demonstrate that MC induces reintroduction of cholestatic injury with TGF-β1.


Methods: Wild-type, KitW-sh (MC-deficient), and Mdr2 – / – mice lacking L-histidine decarboxylase injected with vehicle or PKH26-tagged murine MC by treatment with 0.01% DMSO or inhibitor of TGF-βR, LY2109761 (TGF-βRi, 10 M) three days before the bag. liver damage was assessed by H & E and serum chemistry. MC injected detected in the liver, spleen and lungs by immunofluorescence (IF). DR is measured by CK-19 immunohistochemistry (IHC) and F4 / 80 staining coupled with qPCR for IL-1β, IL-33 and F4 / 80; aging bile evaluated by IF or qPCR for p16, p18 and p21. Fibrosis was evaluated by Sirius Red / Fast Green staining and IF to SYP 9, desmin, and α-SMA. secretion of TGF-β1 / expression was measured by EIA and qPCR. Angiogenesis is detected by the IF to vonWillebrand Factor and VEGF-C qPCR. In vitro, MC TGF-β1 expression / secretion was measured after treatment of TGF-βRi; conditioned media is collected. Cholangiocytes and hepatic stellate cell (HSCs) were treated with conditioned medium and bile MC proliferation / aging is measured by MTS and qPCR; HSC activation was evaluated for α-SMA, SYP-9 and expression of collagen type-1a.


Results: MC injection recapitulation cholestatic liver injury is characterized by an increase in DR; fibrosis / TGF-β1 secretion; and angiogenesis. Injection MC-TGF-βRi reverse this parameter. In vitro, induces gall MC proliferation / activation of HSC aging and overturned by MC less TGF-β1.


Conclusion: Our new research shows that the reintroduction of cholestatic liver injury mimics MC and MC derived TGF-β1 may be a target in chronic cholestatic liver disease.

Mast cells (MCs) induce ductular reaction mimicking liver injury in mice via MC-derived TGF-β1 signaling
Mast cells (MCs) induce ductular reaction mimicking liver injury in mice via MC-derived TGF-β1 signaling

Relieves trimetazidine hypoxia / reoxygenation-induced apoptosis in neonatal rat cardiomyocytes through up-regulating the expression of HMGB1 to promote autophagy

Previous studies have shown the effects of trimetazidine (TMZ) on reducing cardiomyocyte hypoxia / reoxygenation (H / R) injury and the protective effect of autophagy on Ischemia-Reperfusion (I / R) injury to cells. However, whether TMZ protection mechanisms are also involved in autophagy remains unclear.

Our study introduces the role of HMGB1 to check TMZ on autophagy regulation on cardiomyocyte injury H / R. After cell extraction and identification by anti-α-actin staining, cardiomyocytes created hypoxia and reoxygenated, each for 3 hours, and then treated with various concentrations TMZ and transfected with siHMGB1. cell viability and apoptosis were measured by MTS method and flow cytometry, respectively. Autophagy- related factor expression (LC3-I, LC3-II, Beclin-1) and HMGB1 was detected by Western blot and qPCR. Lactate dehydrogenase (LDH) release was assessed by ELISA kit. Cardiomyocytes extracted. H / R lowering of cell viability and increase the level of LDH and cardiomyocyte apoptosis. TMZ had no effect on cardiomyocytes not treated, but reversed the adverse effects of H / R in cardiomyocytes.

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S3200-020 1kit
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Silver protein for bodian staining 7.5-8.5% Ag
S03380 25G
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SDS PAGE Kit (Teaching)
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BioLit Gene Amplification Kit - PCR (Teaching)
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HiPer® Plasmid DNA Extraction Teaching K
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HiPer® Antibody Capture ELISA Teaching K
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HiPer® Bacterial Growth Curve Teaching K
HTM001E-10PR 1 unit
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SpinCol Plasmid Miniprep Kit From Bacterial cells (Teaching)
52166 20 expt. Kit
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Description: Part E

SpinCol Plasmid Miniprep Kit From Bacterial cells (Teaching)
52166-1 50 expt. Kit
EUR 60.64
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Bacterial Transformation Kit – 2 (with blue white screening) (Teaching)
28660 15 expt. Kit
EUR 89.55
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Bacterial Transformation Kit – 2 (with blue white screening) (Teaching)
28660-1 30 expt. Kit
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Silver Stain kit
AR0171 1 kit (for 30 assays to stain the gel of 5 X8.5)
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ElectroPure„¢ Silver Stain Kit
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abx097208-1Kit 1 Kit
EUR 276

HE Staining Kit
E-IR-R117-100mL 100mL
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Hoechst 33342 Staining Kit
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Cell Cycle Staining Kit
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Cell Cycle Staining Kit
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EUR 119
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Cell Cycle Staining Kit
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Sterling Rapid Silver Stain Kit - 1KIT
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CellSurface FLOW Staining Kit
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K2238-100 100T
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Description: Cell Biology|Cell Apoptosis

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Description: Cell Biology|Cell Apoptosis

Beta Galactosidase Staining Kit
55R-1541 250 assays
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Stayrightâ„¢ Purple HRP Staining Kit
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Stayrightâ„¢ Purple HRP Staining Kit
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Stayrightâ„¢ Purple HRP Staining Kit
45906 50 mL
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Stayrightâ„¢ Purple HRP Staining Kit
45906-50mL 50 mL
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Hematoxylin and Eosin Staining Kit
K1142-100 100mL
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Description: Protein & Immunology|Immunofluorescence & Immunohistochemistry

The expression of LC3-II, Beclin-1, and HMGB1 and the ratio of LC3-II / LC3-I increased in H / cardiomyocytes R-processed and subsequently appointed by TMZ pretreatment. However, transfection siHMGB1 exacerbate the effects of H / R in cardiomyocytes and suppresses the protective effects of TMZ on the H / R defective cardiomyocytes with increasing levels of LDH and apoptosis and reduced survival of cardiomyocytes.

Gina

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