Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Morphological characterization of bone marrow-derived mesenchymal stem cells. A: The morphological appearance of bone marrow-derived mesenchymal stem cells (BMSCs) at passage 4 of cell culture (magnification 40 ×); B-F: Expression of green fluorescence protein 72 h after BMSCs transfection by Ad-uPA at different levels of multiplicity of infection (MOI) (MOI = 20, 40, 60, 80, and 100; magnification 40 ×).

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Derivative Assay, Cell Culture, Expressing, Fluorescence, Transfection, Infection

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Western blot detection of urokinase plasminogen activator protein expression in bone marrow-derived mesenchymal stem cells after Ad- urokinase plasminogen activator transfection. Lane 1, uninfected bone marrow-derived mesenchymal stem cells (BMSCs); Lane 2, BMSCs infected with recombinant adenovirus containing human urokinase plasminogen activator (uPA) cDNA; Lane 3, BMSCs infected with control adenovirus. GAPDH: Glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Western Blot, Expressing, Derivative Assay, Transfection, Infection, Recombinant, Control

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Comparison of functional liver levels and liver fibrosis parameters in serum

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Comparison, Functional Assay, Control

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Histopathological change of liver tissue in different groups. A: HE and Masson staining was used to detect structural changes in liver tissues; B: Quantitative analyzes of liver fibrosis were performed using Masson stained sections. Five random views from each sample in each group were analyzed. BMSCs: Bone marrow-derived mesenchymal stem cells; uPA: Urokinase plasminogen activator.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Staining, Derivative Assay

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Effect of urokinase plasminogen activator gene modified-bone marrow-derived mesenchymal stem cells transplantation on the mRNA expression of molecules of the Wnt signaling pathway. The bar graph shows mean relative mRNA expression levels of β-catenin, Wnt4 and Wnt5a in liver tissues. Each sample was repeated three times from each cluster. Data are normalized to GAPDH mRNA expression levels. GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; BMSCs: Bone marrow-derived mesenchymal stem cells; uPA: Urokinase plasminogen activator.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Modification, Derivative Assay, Transplantation Assay, Expressing

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Effect of urokinase plasminogen activator gene modified-bone marrow-derived mesenchymal stem cells transplantation on the protein expression of molecules of the Wnt signaling pathway. A: Representative Western blot analysis of uPA, β-catenin, Wnt4, and Wnt5a protein expression levels in four groups; B: The bar graph represents mean relative protein expression levels of urokinase plasminogen activator, β-catenin, Wnt4, and Wnt5a. Data are normalized to GAPDH protein expression levels. GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; BMSCs: Bone marrow-derived mesenchymal stem cells; uPA: Urokinase plasminogen activator.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Modification, Derivative Assay, Transplantation Assay, Expressing, Western Blot

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Animal experimental flow chart. Exo miR-23b : exosomes derived from BMSCs transfected with miR-23b; Exo miR-NC : exosomes derived from BMSCs transfected with miR-NC; GSH: reduced glutathione; GSSG: oxidized glutathione disulfide; HE: hematoxylin-eosin; ICH: intracerebral hemorrhage; MDA: malondialdehyde; PBS: phosphate-buffered saline; ROS: reactive oxygen species; RT-qPCR: reverse transcription quantitative polymerase chain reaction; SOD: superoxide dismutase; TUNEL: terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: Derivative Assay, Transfection, Saline, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction, TUNEL Assay, End Labeling

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Identification of exosomes derived from BMSCs overexpressing miR-23b. (A) MiR-23b expression in BMSCs detected by RT-qPCR after lentivirus transfection. (B) Transmission electron microscope scanning images of exosomes. The exosomes derived from the BMSCs showed a round cup-shaped and complete structure. Scale bars: 100 nm. (C) Western blot bands detecting exosome markers CD63, TSG101, and CD81. (D) MiR-23b expression levels in different exosome groups detected by RT-qPCR. Data are shown as the mean ± SD ( n = 3). * P < 0.05 (Student’s t -test). BMSCs: Bone marrow mesenchymal stem cells; Exo miR-23b : exosomes derived from BMSCs transfected with miR-23b; Exo miR-NC : exosomes derived from BMSCs transfected with miR-NC; RT-qPCR: reverse transcription quantitative polymerase chain reaction.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Transfection, Transmission Assay, Microscopy, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Exosomal miR-23b inhibits oxidative stress in microglia BV2 cells in vitro . (A) miR-23b levels in microglia BV2 cells after administration of different exosome groups under hemin stimulation. (B) Representative images of ROS staining. The hemin group showed more ROS staining than that in the PBS group, and antioxidant NAC administration further reduced the ROS staining compared with that in the hemin group. ROS levels were decreased in both exomiR-23b and exomiR-NC groups compared with those in the hemin group, and the exomiR-23b group showed less ROS staining than that in the exomiR-NC group. Scale bars: 50 μm. (C) Quantification of relative fluorescence intensity of ROS. (D) MDA levels were measured by economical kits. (E) SOD levels were measured by economical kits. (F) The GSH/ GSSG ration was calculated on the basis the GSH and GSSG levels which evaluated by economical kits. Data are shown as the mean ± SD ( n = 6). $ P < 0.05, $$ P < 0.01, vs . PBS group; * P < 0.05, ** P < 0.01, vs . hemin model group; # P < 0.05, vs . exomiR-NC group (one-way analysis of variance followed by Newman-Keuls post hoc analysis). PBS: Control; PBS + NAC: treated with 1 mM N-acetylcysteine; Hemin: treated with 60 μM hemin for 24 hours; Hemin + NAC: treated with 1 mM N-acetylcysteine 2 hours after hemin stimulation; exomiR-NC group: treated with miR-NC transfected BMSCs-exosomes after hemin stimulation; exomiR-23b group: treated with miR-23b transfected BMSCs-exosomes after hemin stimulation. BMSC: Bone marrow mesenchymal stem cell; GSH: reduced glutathione; GSSG: oxidized glutathione disulfide; ICH: intracerebral hemorrhage; MDA: malondialdehyde; NAC: N-acetylcysteine; PBS: phosphate-buffered saline; ROS: reactive oxygen species; SOD: superoxide dismutase.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: In Vitro, Staining, Fluorescence, Control, Transfection, Saline

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: Exosomal miR-23b alleviates NLRP3 inflammasome-mediated pyroptosis in microglia BV2 cells and protects neuronal cells in vitro . (A, B) Expression and data analysis of pyroptosis-related proteins in different groups of microglia BV2 cells. (C, D) IL-1β (C) and IL-18 (D) levels in microglia BV2 cells were evaluated by ELISA. (E) Cell viabilities of hippocampal neuronal HT22 cells by CCK8 tests. (F) Cell death rates of HT22 cells by LDH assays. Data are shown as the mean ± SD ( n = 6). $$ P < 0.01, vs . PBS group; * P < 0.05, ** P < 0.01, vs . hemin group; # P < 0.05, ## P < 0.01, vs . exo miR-NC group (one-way analysis of variance followed by Newman-Keuls post hoc analysis). PBS group: Control; Hemin group: treated with 60 μM hemin for 24 hours; exo miR-NC group: treated with miR-NC transfected BMSCs-exosomes after hemin stimulation; exo miR-23b group: treated with miR-23b transfected BMSC-exosomes after hemin stimulation. BMSC: Bone marrow mesenchymal stem cell; CCK8: cell counting kit-8; ELISA: enzyme linked immunosorbent assay; GSDMD-N: N-terminal fragment of gasdermin D; IL-18: interleukin-18; IL-1β: interleukin-1β; LDH: lactate dehydrogenase; NLRP3: NOD-like receptor family pyrin domain containing 3; PBS: phosphate-buffered saline.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: In Vitro, Expressing, Enzyme-linked Immunosorbent Assay, Control, Transfection, Cell Counting, Saline

Journal: Neural Regeneration Research

Article Title: Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage

doi: 10.4103/1673-5374.346551

Figure Lengend Snippet: BMSC-exosomal miR-23b attenuates oxidative stress by regulating the PTEN/Nrf2 pathway in microglia BV2 cells in vitro . (A) Western blot bands for PTEN, Nrf2 (cytoplasmic and nuclear), and HO-1 proteins in microglia BV2 cells from different groups. (B) Quantitative analysis of PTEN, Nrf2 (cytoplasmic and nuclear), and HO-1. (C) MDA levels were measured by economical kits. (D) SOD levels were measured by economical kits. (E) GSH/GSSG ratio was calculated on the basis of the GSH and GSSG levels which evaluated by economical kits. Data are shown as the mean ± SD ( n = 6). * P < 0.05, ** P < 0.01, vs . hemin group; # P < 0.05, vs . exo miR-23b group (one-way analysis of variance followed by Newman-Keuls post hoc analysis). Hemin: Treated with 60 μM hemin for 24 hours; exo miR-23b group: treated with miR-23b transfected BMSCs-exosomes after hemin stimulation; exo miR-23b + PTEN: treated with pcDNA3.1-PTEN plasmids and miR-23b transfected BMSCs-exosomes after hemin stimulation. BMSC: Bone marrow mesenchymal stem cell; GSH: reduced glutathione; GSSG: oxidized glutathione disulfide; HO-1: heme oxygenase-1; MDA: malondialdehyde; Nrf2: nuclear factor erythroid-2-related factor 2; PTEN: phosphatase and tensin homolog deleted on chromosome 10; SOD: superoxide dismutase.

Article Snippet: Wistar rat BMSCs (Cat# 170908I32) that had been isolated and cultured were obtained from Cyagen Biosciences Inc. (Santa Clara, CA, USA), and microglia BV2 cells (Cat# 1101MOU-PUMC000063) and hippocampal neuronal HT22 (Cat# BNCC338235) were obtained from Cell Resource Center of the Peking Union Medical College and the Bena Culture Collection (Xinyang, China).

Techniques: In Vitro, Western Blot, Transfection