Journal: Neural Regeneration Research

Article Title: Maintaining moderate levels of hypochlorous acid promotes neural stem cell proliferation and differentiation in the recovery phase of stroke

doi: 10.4103/1673-5374.392889

Figure Lengend Snippet: Clearance of accumulated hypochlorous acid protects from acute ischemic stroke. (A, B) Representative images and quantitative analysis of cerebral infarct size detected by TTC staining in a rat MCAO model following intraperitoneal administration of 50 mg/kg Tau and 30 mg/kg 4-ABAH ( n = 5). Tau and 4-ABAH significantly reduced the infarct area in MCAO rats. White region indicates the infarct area. (C, D) Representative images and statistical analysis of BBB permeability determined by EB staining ( n = 3). Tau and 4-ABAH significantly reduced EB extravasation in MCAO rats. (E) Effect of different concentrations of hypochlorous acid on cleaved caspase-3 expression in HT22 cells under O/R conditions. Hypochlorous acid was administrated upon reoxygenation for 24 hours ( n = 3). (F) Effect of Tau on hypochlorous acid (200 μM)-induced caspase-3 in HT22 cells under O/R conditions ( n = 3). (G, H) Effect of Tau on hypochlorous acid-induced apoptosis in HT22 cells under O/R conditions. Apoptosis was observed by Hoechst 33258 staining ( n = 3). Hypochlorous acid (200 μM) increased apoptosis rate, while Tau (200 μM) reversed the pro-apoptotic effect of hypochlorous acid. Tau markedly inhibited apoptosis in O/R-challenged HT22 cells. Scale bar: 100 μm. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (B, D, E: one-way analysis of variance followed by Dunnett’s post hoc test; F, H: one-way analysis of variance followed by Tukey’s post hoc test). 4-ABAH: 4-Aminobenzohydrazide; EB: Evans blue; MCAO: middle cerebral artery occlusion; O/R: oxygen-glucose deprivation/reoxygenation; Tau: taurine; TTC: 2,3,5-triphenyl-2H-tetrazolium chloride.

Article Snippet: The mouse NSC line C17.2 (kindly provided by Professor Jiangang Shen, RRID: CVCL_4511), microglia cell line BV2 (Cat# 305156, CLS Cell Lines Service GmbH, Eppelheim, Germany, RRID: CVCL_0182), and hippocampal neuronal cell line HT22 (Cat# 305158, CLS Cell Lines Service GmbH, RRID: CVCL_0321) were cultured in complete high-glucose DMEM containing 10% fetal bovine serum and 1% penicillin/streptomycin.

Techniques: Staining, Permeability, Expressing

Journal: Neural Regeneration Research

Article Title: Maintaining moderate levels of hypochlorous acid promotes neural stem cell proliferation and differentiation in the recovery phase of stroke

doi: 10.4103/1673-5374.392889

Figure Lengend Snippet: Moderate hypochlorous acid is beneficial for maintaining NSC proliferation in the recovery phase of stroke. (A) Timeline of the experimental design. Rats were subjected to 2 hours focal ischemia, followed by reperfusion for 10 days. On the 5 th day, 5 μM/10 μL hypochlorous acid and 5 μM/10 μL HKOCl-3 were injected into the lateral ventricle. (B–G) Immunostaining and statistical analysis of Nestin + (Alexa Fluor 488, green) and Sox2 + (Cy3, red) NSCs in ischemic SVZ. Hypochlorous acid decreased Nestin + and Sox2 + intensity on the 7 th day, but increased their intensity on the 10 th day. Data were normalized to the MCAO group. (H) Effect of different concentrations of hypochlorous acid on C17.2 cell viability measured by CCK8 assay. (I, J) BrdU staining (Cy3, red) examined the effect of hypochlorous acid on C17.2 cell proliferation under O/R conditions. The ratio of BrdU + to DAPI + cells was increased by 5 μM hypochlorous acid in O/R-challenged HT22 cells, but inhibited by 200 μM hypochlorous acid. Data are expressed as mean ± SD ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (D, G: one-way analysis of variance followed by Tukey’s post hoc test, H, J: one-way analysis of variance followed by Dunnett’s post hoc test). BrdU: 5-Bromo-2′-deoxyuridine; CCK8: cell counting kit 8; DAPI: 4,6-diamidino-2-phenylindole; NSCs: neural stem cells; O/R: oxygen-glucose deprivation.

Article Snippet: The mouse NSC line C17.2 (kindly provided by Professor Jiangang Shen, RRID: CVCL_4511), microglia cell line BV2 (Cat# 305156, CLS Cell Lines Service GmbH, Eppelheim, Germany, RRID: CVCL_0182), and hippocampal neuronal cell line HT22 (Cat# 305158, CLS Cell Lines Service GmbH, RRID: CVCL_0321) were cultured in complete high-glucose DMEM containing 10% fetal bovine serum and 1% penicillin/streptomycin.

Techniques: Injection, Immunostaining, CCK-8 Assay, BrdU Staining, Cell Counting

Journal: Neural Regeneration Research

Article Title: Inhibiting SHP2 reduces glycolysis, promotes microglial M1 polarization, and alleviates secondary inflammation following spinal cord injury in a mouse model

doi: 10.4103/NRR.NRR-D-23-01925

Figure Lengend Snippet: SHP2 inhibition reduces neuronal apoptosis in the context of SCI. (A, B) NeuN (red-CoraLite594) and SHP2 (green-CoraLite488) immunoreactivities in mice treated with NSC87877 14 days after SCI, as detected by immunofluorescence. Quantitative analysis showing that a decrease in SHP2 fluorescence intensity and an increase in the fluorescence intensity of NeuN, a neuronal marker, in SCI + NSC87877 group mice near the injury site ( n = 5 slices from three mice per group). Scale bars: 50 μm. (C) Nissl staining of mouse spinal cord transverse and longitudinal sections 14 days after SCI showing that, compared with the SCI group, the SCI + NSC87877 group exhibited more Nissl bodies with a more uniform distribution ( n = 5 slices from three mice per group). Scale bars: 100 μm (left), 600 μm (right). (D, E) Western blot analysis of Caspase3, C-caspase3, Bax, and Bcl-2 protein expression in PC12 cells co-cultured with BV2 cells treated with LPS or LPS + NSC87877 for 24 hours. The experiment was repeated three times. (F, G) C-caspase3 (red-CoraLite594) expression in PC12 cells was detected by immunofluorescence staining. Quantitative analysis showed decreased immunofluorescence intensity of the apoptosis marker C-caspase3 in neurons co-cultured with BV2 cells from the LPS + NSC87877 group. The experiment was repeated three times. Scale bars: 50 μm. Data are expressed as the mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance with Bonferroni’s post hoc test). Bax: Bcl-2-associated X protein; Bcl-2: B-cell lymphoma 2; Caspase3: cysteine-aspartic acid protease 3; C-caspase3: cleaved caspase-3; DAPI: 4′,6-diamidino-2′-phenylindole; IF: immunofluorescence staining; LPS: lipopolysaccharide; NeuN: neuronal nuclei; NSC87877: SHP2 inhibitor; SCI: spinal cord injury; SHP2: Src homology 2 domain-containing protein tyrosine phosphatase 2.

Article Snippet: The PC12 neuronal cell line (Cat# CL-0481, RRID:CVCL_0481), and the BV2 mouse microglia cell line (Cat# CL0493, RRID: CVCL_0182) were purchased from Procell Life Science and Technology Co. (Wuhan, China).

Techniques: Inhibition, Immunofluorescence, Fluorescence, Marker, Staining, Western Blot, Expressing, Cell Culture

Journal: Neural Regeneration Research

Article Title: Nanomaterials-mediated lysosomal regulation: a robust protein-clearance approach for the treatment of Alzheimer’s disease

doi: 10.4103/NRR.NRR-D-23-01736

Figure Lengend Snippet: Application of nanomaterials in AD therapy

Article Snippet: Liposomal NPs , 25–10000 , Spherical structure, phospholipid bilayer , Nontoxic, nonimmunogenic, easy to target , Aβ clearance, acetylcholine lipase inhibition , Intravenous injection/Sprague-Dawley rats, bEnd.3 cells, primary rat glia, primary rat neuronal cells , WO 2014 076709A1 , Conjugated to cell-penetrating peptides such as TAT, pVec, and QL , Agrawal et al., 2017; Dos Santos Rodrigues et al., 2019.

Techniques: Inhibition, Injection, Polymer, Antioxidant Activity Assay