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
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.
Description: Abbkine Cell Cycle Staining Kit provides a convenient and accurate determination of the percentage of cells in each phase of the cell cycle.
Description: Abbkine Cell Cycle Staining Kit provides a convenient and accurate determination of the percentage of cells in each phase of the cell cycle.
Description: 3,3'-Diaminobenzidine (DAB) has been applied for decades as the most commonly used IHC chromogen because it is inexpensive and sensitive for routine applications.
Description: 3,3'-Diaminobenzidine (DAB) has been applied for decades as the most commonly used IHC chromogen because it is inexpensive and sensitive for routine applications.
Description: Senescence β-galactosidase staining kit provides a simple and convenient operation method for staining detection of senescent cells or tissues; This kit only stains senescent cells, and does not stain pre-senescence cells (senescent cells), quiescent cells (static cells), immortal cells (immortal cells) or tumor cells.
Description: Alkaline Phosphatase (AP) is a widely used marker for both mouse and human embryonic stem cells (ES) and embryonic germ cells (EG). Our StemTAG Alkaline Phosphatase kits provide an efficient system for monitoring cell differentiation or undifferentiation using the AP marker. The StemTAG Alkaline Phosphatase Staining Kits provide reagents for monitoring alkaline phosphatase activity via immunocytochemistry staining.
Description: Gelite™ Orange is an extremely sensitive nucleic acid gel stain for detecting DNA or RNA in gels using a standard 300 nm UV transilluminator and Polaroid 667 black-and-white print film.
Description: Alkaline Phosphatase (AP) is a widely used marker for both mouse and human embryonic stem cells (ES) and embryonic germ cells (EG). Our StemTAG Alkaline Phosphatase kits provide an efficient system for monitoring cell differentiation or undifferentiation using the AP marker. The StemTAG Alkaline Phosphatase Staining Kits provide reagents for monitoring alkaline phosphatase activity via immunocytochemistry staining.
Description: Cell Navigator® Live Cell Tubulin Staining Kit provides a robust method to fluorescently image tubulins in live cells with Tubulite™ Red.
Description: Cell Navigator® Live Cell Tubulin Staining Kit provides a robust method to fluorescently image tubulins in live cells with Tubulite™ Red.
Description: Double-strand breaks (DSB) in DNA are among the most dangerous types of DNA damage occuring within cells. One of the earliest cellular responses to double-strand breaks is the phosphorylation of a histone variant, H2AX, at the sites of DNA damage. Within seconds Ser139 is phosphorylated when DSBs are induced in mammalian cells. Phosphorylation of this serine residue causes chromatin condensation and appears to play a critical role in the recruitment of repair or damage-signaling factors to the DNA damage sites. The OxiSelect DNA Double-Strand Break Staining Kit provides an easy-to-use method for detecting the presence of DSBs in cells cultured in microtiter plates. Double strand breaks can be detected in just a few hours by immunofluorescence staining of the phosphorylated histone H2AX.
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.
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