Journal: Neural Regeneration Research

Article Title: Hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect neurons from cardiac arrest–induced pyroptosis

doi: 10.4103/NRR.NRR-D-23-01922

Figure Lengend Snippet: PFKL mediates the neuroprotective and antioxidant effects of HP-BMSCs in CPR rats. (A) Neurological deficit scores (NDS) in four group (CPR + PBS, CPR + HP-BMSCs, CPR + si NC HP-BMSCs, CPR + si-PFKL HP-BMSCs, n = 8). The black line represents the NDS levels in the sham group. (B) Serum S100B levels after treatment as indicated ( n = 8). (C) Nissl staining of rat brains after CPR in different group (sham, CPR + PBS, CPR + HP-BMSCs, CPR + si NC HP-BMSCs, CPR + si-PFKL HP-BMSCs, n = 8). After 24 hours of resuscitation, the number of Nissl bodies in the rat cortex significantly decreased, and transplantation of HP-BMSCs could alleviate the reduction of Nissl bodies. However, this effect of HP-BMSCs was reversed after knocking down PFKL. Scale bars: 100 μm (upper) or 50 μm (lower). (D) The AOD of cortical neurons by Nissl staining. (E, F) Representative immunohistochemical staining of 4-HNE (E) and 8-OHdG (F) in cortex following ROSC. After transplantation of HP-BMSCs, the expression of 4-HNE and 8-OHdG in the cortex of CA/CPR rats significantly decreased, whereas this effect was attenuated after knocking down PFKL. The selected areas (black frame) were displayed at a higher magnification level. Scale bars: 70 μm (upper) or 50 μm (lower). (G, H) Percentage of 4-HNE-positive cells (G) and 8-OHdG-positive cells (H) in immunohistochemical images ( n = 8). (I–K) Histograms show the expression of SOD (I), GSH-Px (J) and MDA (K). n = 6 rats per group. (L) Representative images show iNOS (green, Alexa Fluor 488) immunostaining with NeuN (red, CY3) and DAPI (blue) in the cotex ( n = 8). The white boxed areas are shown magnified. HP-BMSCs significantly reduced the expression of iNOS in the neurons of CA-induced rats, while knocking down the expression of PFKL in HP-BMSCs did not lead to significant changes in the expression of iNOS. Scale bars: 50 μm (lower) or 1000 μm (upper). (M) Percentage of iNOS-positive neurons in immunofluorescence images. (N) ROS generation in cortex post-resuscitation was assayed by ROS assay kit. Compared with the si-NC HP-BMSCs group, the reduction effect of si-PFKL HP-BMSCs on post-resuscitation cortical ROS is significantly weakened. The fluorescence was detected by the fluorescence microplate reader ( n = 3). Scale bars: 100 μm. (O) The fluorescent intensity of ROS shown in N. Results expressed as median and interquartile range (A and D) or means ± SD (G–K, M and O). * P < 0.05, vs . sham group; # P < 0.05, vs . CPR + PBS group; & P < 0.05, vs . CPR + si-NC-BMSCs group (Kruskal–Wallis test followed by Dunn’s multiple comparisons test in A and D; one-way analysis of variance followed by Tukey’s multiple comparisons test in G–K, M and O). 4-HNE: 4-Hydroxynonenal; 8-OHdG: 8-hydroxy-2′-deoxyguanosine; AOD: average optical density; CPR: cardiopulmonary resuscitation; HP-BMSCs: hypoxia preconditioned bone marrow mesenchymal stem cells; iNOS: inducible nitric oxide synthase; MDA: malondialdehyde; PBS: phosphate-buffered saline; PFKL: liver isoform of phosphofructokinase; ROI: region of interest; ROS: reactive oxygen species; ROSC: return of spontaneous circulation; SOD: superoxide dismutase.

Article Snippet: For the second experiment, the cells were divided into five groups that were left untreated or treated with siRNAs, left untreated or treated with 10 μM mitochondria-targeted antioxidant (Mito-TEMPO, TargetMol, Shanghai.

Techniques: Staining, Transplantation Assay, Immunohistochemical staining, Expressing, Immunostaining, Immunofluorescence, ROS Assay, Fluorescence, Analysis, Saline, Immunohistochemistry, Knockdown

Journal: PLOS ONE

Article Title: Clinical outcomes and anti-inflammatory mechanisms predict maximum heart rate improvement after physical activity training in individuals with psychiatric disorders and comorbid obesity

doi: 10.1371/journal.pone.0313759

Figure Lengend Snippet: MHR: maximum heart rate, HRSD: Hamilton rating scale for depression, PANSS-P: positive and negative syndrome scale (positive subscale), PANSS-G: positive and negative syndrome scale (general subscale), IL-10: interleukin 10, TNF-α: tumor necrosis factor alpha, ROS: reactive oxygen species; ΔΨm: mitochondrial membrane potential, VCAM: vascular cellular adhesion molecule, LEPMN: leukocyte-endothelium adhesion polymorphonuclear, RVPMN: rolling velocity polymorphonuclear cells, PSEL: P-selectin; MPO: myeloperoxidase.

Article Snippet: In total, 100 000 cells were plated in each well of 96-well plates and incubated for 30 min at 37°C with the corresponding fluorochromes, as follows: dichlorofluorescein diacetate to measure reactive oxygen species (ROS) production (485 nm excitation, 535 nm emission), MitoSOX to measure mitochondrial ROS (mROS) (510 nm excitation, 580 nm emission), tetramethylrodamin methyl ester to assess mitochondrial membrane potential (ΔΨm) (552 nm excitation, 574 nm emission), superoxide dismutase (SOD) (total SOD/T-SOD Activity Assay Kit–Colorimetric—from NOVUS), and 5-chloromethylfluorescein diacetate to measure intracellular glutathione (GSH) (492 nm excitation, 517 nm emission).

Techniques: Membrane