mitochondrion targeted antioxidant mito tempo  (MedChemExpress)

Journal: Hepatology Communications

Article Title: Antioxidant interventions reduced cytokine-induced pyroptosis of peripheral MAIT cells in patients with HBV-related cirrhosis

doi: 10.1097/HC9.0000000000000714

Figure Lengend Snippet: Antioxidants reduced the cytokine-induced pyroptosis of mucosal-associated invariant T (MAIT) cells. (A–G) Peripheral blood mononuclear cells from healthy controls, patients with compensated liver cirrhosis, and patients with decompensated liver cirrhosis were treated with IL-18 or IL-12 stimulants with polyphenols, MTA, or iACAT for 24 hours, respectively. The levels of DCFH-DA MFI, FLICA caspase-1, IFN-γ, TNF-a, GZMB, CD107a, and IL-17A expressed in the MAIT cells were determined by flow cytometry. (A–G) Wilcoxon signed-rank test. * p <0.05, *** p <0.001, and **** p <0.0001, ns, non-signifcant. Abbreviations: MFI, mean fluorescence intensity; polyphenols, resveratrol and oleuropein; MTA, Mito-TEMPO and MitoQ; iACAT, Avasimibe and K-604.

Article Snippet: For the intervention with antioxidants, the treatments included the following: (1) medium only, (2) the addition of IL-12 or IL-18 at a concentration of 200 ng/mL, and (3) the same combination supplemented separately with polyphenols (ie, resveratrol [5 μM, APExBIO] and oleuropein [0.25 μM, APExBIO]); iACAT (ie, avasimibe [0.25 μM, Selleck], and K-604 [0.05 μM, MCE]; and mitochondria-targeted antioxidants (MTA) (ie, Mito-TEMPO [5 μM, Merk] and MitoQ [0.05 μM, MCE]).

Techniques: Flow Cytometry, Fluorescence

Journal: Hepatology Communications

Article Title: ER stress promotes mitochondrial calcium overload and activates the ROS/NLRP3 axis to mediate fatty liver ischemic injury

doi: 10.1097/HC9.0000000000000399

Figure Lengend Snippet: Mitochondrial calcium overload aggravates NLRP3 activation by oxidative stress in macrophages. Control and HFD-fed mice were treated with a liver IR model or sham procedure. Intrahepatic macrophages were collected at 6 hours after reperfusion. (A) MDA and GSH/GSSG levels and SOD activity in macrophages. (B) ROS levels were detected by DCFH-DA fluorescence. Control and HFD-fed mice were pretreated with Xestospongin-C (3 μM) or PBS (Control) before ischemia. Intrahepatic macrophages and the liver tissues were collected at 6 hours after reperfusion. (C, D) ROS levels were detected by DCFH-DA fluorescence and 4-HNE. Control and HFD-fed mice were pretreated with mito-TEMPO or PBS (Control) before ischemia. Intrahepatic macrophages were collected at 6 hours after reperfusion. (E) Western blot analysis of NLRP3, C-caspase-1, Pro-caspase-1, IL-1β, IL-18, and GAPDH in isolated macrophages. (F) H&E staining of liver sections. n=4 mice/group. Values were presented as the mean±SD. * p <0.05. Abbreviations: 4-HNE, 4-hydroxynonenal; DCFH-DA, 2,7-dichlorodi-hydrofluorescein diacetate; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GSH, reduced glutathione; GSSG, oxidized glutathione disulfide; H&E, hematoxylin-eosin; HFD, high-fat diet; IR, ischemia and reperfusion; MDA, malondialdehyde; NLRP3, NOD-like receptor thermal protein domain–associated protein 3; ROS, reactive oxygen species; SOD, superoxide dismutase.

Article Snippet: To determine the role of mitochondrial oxidation, Control and HFD-fed mice were pretreated with the mitochondria-targeted antioxidant Mito-TEMPO (5 mg/kg, MedChemExpress, HY-112879) twice at 17 hours and 1 hour before surgery or PBS.

Techniques: Activation Assay, Control, Activity Assay, Fluorescence, Western Blot, Isolation, Staining

Journal: Biomedicines

Article Title: Aldosterone Suppresses Endothelial Mitochondria through Mineralocorticoid Receptor/Mitochondrial Reactive Oxygen Species Pathway

doi: 10.3390/biomedicines10051119

Figure Lengend Snippet: Mito-TEMPO reduces the function of mitochondrial in the aldosterone-stimulated HUVECs. HUVECs were treated with Mito-TEMPO (10 uM) for 2 h before being treated with 10 −7 M aldosterone. ( A ) After 24 h, immunofluorescence images showed mtROS production. ( B ) HUVECs were transfected with GR siRNA (50 nM) or MR siRNA (100 nM) for 6 h and then treated with 10 −7 M aldosterone. MtROS was stained red with MitoSox, and mitochondria were stained green with MitoTracker. ( C ) The mitochondrial DNA Cyt b copy number (up) and mitochondrial DNA COII copy number (down) were quantified by qPCR. ( D ) Mitochondria were stained red with anti-mitochondrial antibodies, and cell nuclei were stained blue with DAPI. Representative images were captured using a fluorescence microscope with 40x magnification. Scale bar: 100 μm. Quantitative fluorescence intensity was analyzed using Image J software. * p < 0.05, ** p < 0.01, compared between two groups using the t -test.

Article Snippet: Aldosterone and N-acetyl-L-cysteine ((NAC) an antioxidant) were purchased from Sigma (St Louis, MO, USA), and Mito-TEMPO (a mitochondria-targeted antioxidant) was purchased from Enzo Life Sciences (Farmingdale, NY, USA).

Techniques: Immunofluorescence, Transfection, Staining, Fluorescence, Microscopy, Software