Exosomes Secreted From Bone Marrow Mesenchymal Stem Cells Attenuate Oxygen-Glucose Deprivation/Reoxygenation-Induced Pyroptosis in PC12 Cells by Promoting AMPK-Dependent Autophagic Flux
Background: Cerebral ischemia-reperfusion (I / R) injury can cause severe dysfunction, and treatment difficult. It is reported that the nucleotide-binding domain and a leucine-rich repeat protein family of 3 (NLRP3) inflammasome-mediated cell pyroptosis an important part of the I / R injury of the brain and can inhibit the activation of autophagy pyroptosis in some tissue injury. Our previous research found that the protective effect of bone marrow mesenchymal stem cells (BMSCs) in brain I / R injury may be associated with the regulation of autophagy.
Recent research has shown that exosomes secreted from BMSCs (BMSC-Exos) may play an important role in effective biological performance BMSCs and BMSC-Exos protective mechanism associated with the activation of autophagy and remission of inflammation, but have not been reported in studies of injury I / R brain. We aimed to investigate the effects of BMSC-Exos on I / R injury of the brain and determine whether the mechanisms associated with the regulation pyroptosis and autophagic flux.
Methods: PC12 cells were subjected to a shortage of oxygen-glucose / reoxygenation (OGD / R) to induce brain I / R in vitro and cocultured with BMSC-Exos. cell viability was determined by CCK-8 kit and lactate dehydrogenase (LDH) detection. Scanning electron microscopy (SEM), Hoechst 33 342 / propidium iodide (PI) double staining, 2 ‘, 7’-dichlorodihydrofluorescein diacetate assay, immunofluorescence, Western blot and enzyme-linked immunosorbent assay (ELISA) was used to detect cell pyroptosis. Furthermore, transmission electron microscopy (TEM), GFP-RFP-LC3 adenovirus transfection, and Western blot used to detect autophagic flux and its influence on pyroptosis.
Finally, coimmunoprecipitation used to detect the binding interaction between NLRP3 and LC3. Results: BMSC-Exos increased cell viability in OGD / R inhibitory effect of BMSC-Exos on pyroptosis is comparable to NLRP3 inhibitor NLRP3 MCC950 and reversed by excess. Furthermore, BMSC-Exos promoted autophagic flux through AMP-activated kinase (AMPK) / mammalian target of rapamycin on the track, whereas chloroquine, AMPK silencing, and compound C block the inhibitory effects on pyroptosis. Conclusion: BMSC-Exos can protect PC12 cells against injury OGD / R by the weakening of the NLRP3 inflammasome-mediated pyroptosis to promote the AMPK-dependent autophagic flux.
Exosomes Secreted From Bone Marrow Mesenchymal Stem Cells Attenuate Oxygen-Glucose Deprivation/Reoxygenation-Induced Pyroptosis in PC12 Cells by Promoting AMPK-Dependent Autophagic Flux
Effect of Morinda citrifolia (Noni) on Cell Viability of Stem Cells and Osteogenesis spheroids
Background and purpose: Morinda citrifolia (Noni) has been widely used in herbal medicine to treat and prevent various diseases. We undertook this study to evaluate the effects of Noni extract on morphology maintenance, increase cellular viability, and improved osteogenesis of stem cell spheroids.
Materials and Methods: We cultured stem cell spheroids made with gingival-derived stem cells in the presence of Noni extract at concentrations of 10, 100 and 200 ng / mL. We do the analysis of changes in cell morphology and cell viability. We did a test alkaline phosphatase activity using a kit and test the mineralization using anthraquinone dye to evaluate osteogenesis of stem cell spheroids with the addition of Noni extract.
Description: TRAP Antibody: TRAP, also known as uteroferrin, is an iron containing, glycosylated, acid phosphatase. It is the most basic of the acid phosphatases and is the only form not inhibited by L(+)-tartrate. Along with the related protein ACP2, TRAP mediates the removal of mannose 6-phosphate residues from proteins targeted to lysosomes. TRAP is present in brain at low levels, but is expressed at a much higher level in liver.
Description: TRAP Antibody: TRAP, also known as uteroferrin, is an iron containing, glycosylated, acid phosphatase. It is the most basic of the acid phosphatases and is the only form not inhibited by L(+)-tartrate. Along with the related protein ACP2, TRAP mediates the removal of mannose 6-phosphate residues from proteins targeted to lysosomes. TRAP is present in brain at low levels, but is expressed at a much higher level in liver.
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human TRAP . This antibody is tested and proven to work in the following applications:
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: 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: A polyclonal antibody raised in Rabbit that recognizes and binds to Human TRAP (N-term). This antibody is tested and proven to work in the following applications:
Description: Quantitativesandwich ELISA kit for measuring Human Tartrate Resistant Acid Phosphatase (TRAP) in samples from serum, plasma, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Human Tartrate Resistant Acid Phosphatase(TRAP)ELISA Kit
Description: Quantitativesandwich ELISA kit for measuring Human Tartrate Resistant Acid Phosphatase(TRAP) in samples from serum, plasma, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: Our Cellular Senescence Staining Kit provides an efficient method to visualize Senescence Associated (SA) ß-galactosidase. SA-ß-Gal catalyzes the hydrolysis of X-gal, which produces a blue color in senescent cells. Visualize results with a standard light microscope.
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Results: The cells formed spheroids well applied, and the addition of Noni in 10, 100 and 200 concentration ng / mL did not produce significant morphological changes. Quantitative values for the survival of the mobile on day 3 showed that the values of absorbance at 450 nm is 0.314 ± 0.013, 0.318 ± 0.008, 0.304 ± 0.000 and 0.300 ± 0.011 for Noni at 0, 10, 100 and 200 concentration ng / mL , each.
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