Journal: Aging (Albany NY)

Article Title: miR-183-5p alleviates early injury after intracerebral hemorrhage by inhibiting heme oxygenase-1 expression

doi: 10.18632/aging.103343

Figure Lengend Snippet: miR-183-5p alleviated early inflammation and oxidative damage by directly targeting heme oxygenase-1 (HO-1). ( A ) Above: schematic showing the potential miR-183-5p binding site in the HO-1 3’-untranslated region (3’-UTR). A mutant (MUT) HO-1 3’-UTR was introduced by replacing the wild type (WT) binding sequence with a mutant sequence. Below: inhibition of relative luciferase activity of HO-1 3’-UTR reporter molecules in human embryonic kidney 293 cells mediated by miR-183-5p. miR-183-5p MM, miR-183-5p mimic; NC, nontarget control. ( B ) Above: western blotting revealed that miRNA-183-5p downregulated HO-1 expression. Below: quantitative analysis of HO-1 protein expression in different groups. n = 8/group. ( C ) Quantitative analysis of cytokine expression in the brains of mice from different groups pretreated with the HO-1 inhibitor zinc protoporphyrin IX (ZnPP) at 3 days after ICH. n = 8/group. ( D ) Quantitative analysis of 4-HNE expression in the brains of mice from different groups pretreated with the HO-1 inhibitor ZnPP at 3 days after ICH. n = 8/group. Values are presented as the mean ± standard deviation. * P < 0.05 vs. the ICH group.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-HO-1 (1:100, ADI-OSA-110, Enzo Life Sciences, Inc., Farmingdale, NY, USA), rabbit anti-Iba-1 (1:200, ab178846, Abcam Plc., Cambridge, UK), and rabbit anti-myeloperoxidase (MPO, 1:100, ab9535, Abcam).

Techniques: Binding Assay, Mutagenesis, Sequencing, Inhibition, Luciferase, Activity Assay, Western Blot, Expressing, Standard Deviation

Journal: Aging (Albany NY)

Article Title: miR-183-5p alleviates early injury after intracerebral hemorrhage by inhibiting heme oxygenase-1 expression

doi: 10.18632/aging.103343

Figure Lengend Snippet: miRNA-183-5p affected microglial survival by targeting heme oxygenase-1 (HO-1) after intracerebral hemorrhage (ICH). ( A ) Left: representative immunofluorescence images of HO-1 in Iba-1–positive microglia at 3 days after ICH. Right: percentage of both Iba-1– and HO-1–positive cells in Iba-1–positive microglia. Scale bars = 50 μm, n = 8/group. * P < 0.05 vs. the ICH group. ( B ) Above: representative immunofluorescence images of HO-1 in BV2 microglia from different groups at 24 hours after hemin treatment. Below: percentage of HO-1–positive BV2 microglia. Scale bars = 50 μm, n = 3/group. * P < 0.05 vs. the hemin group. ZnPP, zinc protoporphyrin IX; CoPP, cobalt protoporphyrin IX.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-HO-1 (1:100, ADI-OSA-110, Enzo Life Sciences, Inc., Farmingdale, NY, USA), rabbit anti-Iba-1 (1:200, ab178846, Abcam Plc., Cambridge, UK), and rabbit anti-myeloperoxidase (MPO, 1:100, ab9535, Abcam).

Techniques: Immunofluorescence

Journal: Aging (Albany NY)

Article Title: miR-183-5p alleviates early injury after intracerebral hemorrhage by inhibiting heme oxygenase-1 expression

doi: 10.18632/aging.103343

Figure Lengend Snippet: miR-183-5p regulated heme oxygenase-1 (HO-1) independent of Nrf2. ( A ) Western blotting revealed that miRNA-183-5p is an HO-1–dependent inhibitor of Nrf2 phosphorylation. n = 8/group. ( B ) Quantitative analysis of the relative expression of p-Nrf2 protein in ( A ). ( C ) Quantitative analysis of the HO-1–dependent inhibitory effect of Nrf2 on miR-183-5p by RT-qPCR. n = 8/group. Values are presented as the mean ± standard deviation. * P < 0.05 vs. the intracerebral hemorrhaging (ICH) group. ZnPP, zinc protoporphyrin IX; tBHQ, tert-butylhydroquinone.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-HO-1 (1:100, ADI-OSA-110, Enzo Life Sciences, Inc., Farmingdale, NY, USA), rabbit anti-Iba-1 (1:200, ab178846, Abcam Plc., Cambridge, UK), and rabbit anti-myeloperoxidase (MPO, 1:100, ab9535, Abcam).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Standard Deviation

Journal: Aging (Albany NY)

Article Title: miR-183-5p alleviates early injury after intracerebral hemorrhage by inhibiting heme oxygenase-1 expression

doi: 10.18632/aging.103343

Figure Lengend Snippet: The experimental design. Sham group, sham operation group; ICH group, collagenase-induced intracerebral hemorrhage group; miRNA-seq, miRNA sequencing; miR-183-5p, microRNA-183-5p; HO-1, heme oxygenase-1; ICH 1 d, 3 d, 7 d, 14 d, 28 d groups, 1 day, 3 days, 7 days, 14 days, 28 days after collagenase-induced intracerebral hemorrhage groups; ZnPP, HO-1 inhibitor zinc protoporphyrin IX; Nrf2 -/- , nuclear factor erythroid 2-related factor knockout; CoPP, HO-1 inducer cobalt protoporphyrin IX; tBHQ, Nrf2 activator tert-butylhydroquinone.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-HO-1 (1:100, ADI-OSA-110, Enzo Life Sciences, Inc., Farmingdale, NY, USA), rabbit anti-Iba-1 (1:200, ab178846, Abcam Plc., Cambridge, UK), and rabbit anti-myeloperoxidase (MPO, 1:100, ab9535, Abcam).

Techniques: Sequencing, Knock-Out

Journal: The Journal of Biological Chemistry

Article Title: Biliverdin Reductase A Attenuates Hepatic Steatosis by Inhibition of Glycogen Synthase Kinase (GSK) 3β Phosphorylation of Serine 73 of Peroxisome Proliferator-activated Receptor (PPAR) α *

doi: 10.1074/jbc.M116.731703

Figure Lengend Snippet: Liver-specific knock-out of BVRA in mice increases blood glucose. A, real time PCR of BVRA in the liver of LBVRA KO and flox mice. B, hepatic immunofluorescence staining for BVRA. Scale bars, 75 μm. C, Western blot of BVRA from different tissues. B, brain; H, heart; K, kidney; L, liver; S, spleen. D, hepatic BVR activity. E–I, measurement of body weight (E), lean and fat mass (F), as well as plasma bilirubin (G), glucose (H), and insulin (I) in LBVRA KO and control flox mice. *, p < 0.05 (versus floxed mice; ± S.E.; n = 6–8/group).

Article Snippet: BVRA was detected using a rabbit anti-BVRA antibody (ADI-OSA-450, 1:100; Enzo Life Sciences, Farmingdale, NY).

Techniques: Knock-Out, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Western Blot, Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Biliverdin Reductase A Attenuates Hepatic Steatosis by Inhibition of Glycogen Synthase Kinase (GSK) 3β Phosphorylation of Serine 73 of Peroxisome Proliferator-activated Receptor (PPAR) α *

doi: 10.1074/jbc.M116.731703

Figure Lengend Snippet: BVRA regulates hepatic insulin signaling. A and B, Western blot and densitometry of the insulin receptor precursor and insulin receptor-β (IRβ) in the liver (A) or AKT1/2 and phosphorylated AKT in the liver (B). C, GTT. D, ITT in LBVRA KO and control flox mice. **, p < 0.01 (versus flox mice; ± S.E.; n = 4); ***, p < 0.001 (versus flox mice; ± S.E.; n = 4).

Article Snippet: BVRA was detected using a rabbit anti-BVRA antibody (ADI-OSA-450, 1:100; Enzo Life Sciences, Farmingdale, NY).

Techniques: Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Biliverdin Reductase A Attenuates Hepatic Steatosis by Inhibition of Glycogen Synthase Kinase (GSK) 3β Phosphorylation of Serine 73 of Peroxisome Proliferator-activated Receptor (PPAR) α *

doi: 10.1074/jbc.M116.731703

Figure Lengend Snippet: The loss of hepatic BVRA causes lipid accumulation. A, hepatic fat measured by Echo-MRI. B, biochemical measurement of hepatic triglyceride levels. C, Oil Red O staining of liver sections and densitometry. Scale bars, 50 μm. D–F, representative Western blot of hepatic expression of AMPK and phosphorylated AMPK (D), FAS (E), and phosphorylated ACC (F). *, p < 0.05; **, p < 0.01 (versus flox mice; ± S.E.; n = 4).

Article Snippet: BVRA was detected using a rabbit anti-BVRA antibody (ADI-OSA-450, 1:100; Enzo Life Sciences, Farmingdale, NY).

Techniques: Staining, Western Blot, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Biliverdin Reductase A Attenuates Hepatic Steatosis by Inhibition of Glycogen Synthase Kinase (GSK) 3β Phosphorylation of Serine 73 of Peroxisome Proliferator-activated Receptor (PPAR) α *

doi: 10.1074/jbc.M116.731703

Figure Lengend Snippet: Diagram of hepatic BVRA signaling to reduce fatty liver and increase glycogen storage. BVRA reduces biliverdin to bilirubin, which binds to and activates the nuclear receptor PPARα for the burning of fat by β-oxidation and storage of glycogen by increasing GYS2. Additionally, BVRA binds directly to AKT to increase phosphorylation, which inhibits GSK3β activity to increase glycogen and decrease lipid storage in the liver. The BVRA-AKT interaction also increases hepatic insulin sensitivity and glucose uptake.

Article Snippet: BVRA was detected using a rabbit anti-BVRA antibody (ADI-OSA-450, 1:100; Enzo Life Sciences, Farmingdale, NY).

Techniques: Activity Assay