Research Paper Volume 13, Issue 1 pp 262—278

Combined transplantation of neural stem cells and bone marrow mesenchymal stem cells promotes neuronal cell survival to alleviate brain damage after cardiac arrest via microRNA-133b incorporated in extracellular vesicles

miR-133b in BMSCs-derived EVs mediates the AKT-GSK-3β-WNT pathway by targeting JAK1. (A) Prediction binding site of miR-133b in JAK1 3'-UTR. (B) Detection of luciferase activity using dual-luciferase reporter gene assay; * p vs. the NC group. (C) The expression of JAK1 in neuronal cells normalized to GAPDH in response to miR-133b mimic determined by Western blot analysis; * p vs. the mimic NC group (neuronal cells treated with mimic NC). (D) The expression of JAK1 in neuronal cells normalized to GAPDH in response to treatment of BMSCs-derived EVs determined by Western blot analysis. (E) Protein levels of JAK1, p-AKT/AKT, p-GSK-3β/GSK-3β, and WNT-3 in the neuronal cells normalized to GAPDH in response to treatment of BMSCs-derived EVs determined by Western blot analysis. * p vs. the PBS group (neuronal cells treated with PBS). (F) Protein levels of JAK1, p-AKT/AKT, p-GSK-3β/GSK-3β, and WNT-3 in the neuronal cells normalized to GAPDH in response to treatment of EVs derived from the BMSCs treated with miR-133b inhibitor determined by Western blot analysis. * p vs. the EVs-inhibitor NC group (neuronal cells in co-culture system with EVs derived from the BMSCs treated with inhibitor NC). Data obtained from three independent cell experiments were expressed as mean ± standard deviation. Data between two groups were analyzed by the unpaired t test.

Figure 5. miR-133b in BMSCs-derived EVs mediates the AKT-GSK-3β-WNT pathway by targeting JAK1. (A) Prediction binding site of miR-133b in JAK1 3'-UTR. (B) Detection of luciferase activity using dual-luciferase reporter gene assay; * p < 0.05 vs. the NC group. (C) The expression of JAK1 in neuronal cells normalized to GAPDH in response to miR-133b mimic determined by Western blot analysis; * p < 0.05 vs. the mimic NC group (neuronal cells treated with mimic NC). (D) The expression of JAK1 in neuronal cells normalized to GAPDH in response to treatment of BMSCs-derived EVs determined by Western blot analysis. (E) Protein levels of JAK1, p-AKT/AKT, p-GSK-3β/GSK-3β, and WNT-3 in the neuronal cells normalized to GAPDH in response to treatment of BMSCs-derived EVs determined by Western blot analysis. * p < 0.05, vs. the PBS group (neuronal cells treated with PBS). (F) Protein levels of JAK1, p-AKT/AKT, p-GSK-3β/GSK-3β, and WNT-3 in the neuronal cells normalized to GAPDH in response to treatment of EVs derived from the BMSCs treated with miR-133b inhibitor determined by Western blot analysis. * p < 0.05, vs. the EVs-inhibitor NC group (neuronal cells in co-culture system with EVs derived from the BMSCs treated with inhibitor NC). Data obtained from three independent cell experiments were expressed as mean ± standard deviation. Data between two groups were analyzed by the unpaired t test.