Journal: Pharmacological research

Article Title: Transfer of miR-877-3p via extracellular vesicles derived from dental pulp stem cells attenuates neuronal apoptosis and facilitates early neurological functional recovery after cerebral ischemia-reperfusion injury through the Bclaf1/p53 signaling pathway.

doi: 10.1016/j.phrs.2024.107266

Figure Lengend Snippet: Fig. 3. DPSC-EVs reduce I/RI-induced neuronal apoptosis. A Representative micrographs of HE and Nissl staining in periinfarction cortex (n=3). Scale bars in A represent 100 μm. B-C Quantification of the percentage of Tunel-positive cells (apoptotic cells labeled with Tunel emitted green fluorescence) in different groups after I/RI (n=4). Scale bars in C represent 100 μm. D-E Representative micrographs and quantitative analysis of FJC-positive cells in periinfarction cortex at 24 h after I/RI (n=4). Scale bars in E represent 50 μm. F-I Representative WB bands and densitometric quantification of Bax, Bcl-2, caspase-3, and cleaved-caspase-3 in the ipsi lateral hemisphere at 24 h after I/RI (n=6). All data was represented as mean ± SD. **P<0.01, ***P<0.001 and ****P<0.0001 sham group vs. tMCAO + saline group, respectively; #P<0.05 and ##P<0.01 tMCAO + saline group vs. tMCAO + DPSC-EVs group; two-tailed independent Student’s t-test.

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) assay Apoptotic cells in cortex were identified using a Tunel assay with an apoptosis detection kit (Proteintech, China), following the manufacturer’s instructions to detect nuclear fragmentation.

Techniques: Staining, TUNEL Assay, Labeling, Fluorescence, Saline, Two Tailed Test

Journal: Pharmacological research

Article Title: Transfer of miR-877-3p via extracellular vesicles derived from dental pulp stem cells attenuates neuronal apoptosis and facilitates early neurological functional recovery after cerebral ischemia-reperfusion injury through the Bclaf1/p53 signaling pathway.

doi: 10.1016/j.phrs.2024.107266

Figure Lengend Snippet: Fig. 4. DPSC-EVs inhibit apoptosis and protect neurons from OGD/R injury. A Immunofluorescence staining for DPSC-EVs uptake by SH-SY5Y cells. Scale bars in A represent 20 μm. B-E Representative WB bands and densitometric quantification of Bax, Bcl-2, caspase-3, and cleaved-caspase-3 in periinfarction cortex at 24 h after OGD/R (n=3). F-G Representative micrographs and quantitative analysis of Tunel-positive cells after OGD/R (n=3). All data was represented as mean ± SD. **P<0.01, ***P<0.001 and ****P<0.0001 normoxia group vs. OGD/R + PBS group, respectively; #P<0.05 and ##P<0.01 OGD/R + PBS group vs. OGD/R + DPSC- EVs group; two-tailed independent Student’s t-test. Scale bars in G represent 100 μm.

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) assay Apoptotic cells in cortex were identified using a Tunel assay with an apoptosis detection kit (Proteintech, China), following the manufacturer’s instructions to detect nuclear fragmentation.

Techniques: Immunofluorescence, Staining, TUNEL Assay, Two Tailed Test

Journal: Pharmacological research

Article Title: Transfer of miR-877-3p via extracellular vesicles derived from dental pulp stem cells attenuates neuronal apoptosis and facilitates early neurological functional recovery after cerebral ischemia-reperfusion injury through the Bclaf1/p53 signaling pathway.

doi: 10.1016/j.phrs.2024.107266

Figure Lengend Snippet: Fig. 6. miR-877-3p inhibitor reversed the effects of DPSC-EVs on Bclaf1 protein downregulation and apoptosis inhibition. A-B Representative WB bands and densitometric quantification of Bclaf1 of each animal group (n=6). All data was represented as mean ± SD. ***P<0.001 sham group vs. tMCAO + saline group; ###P<0.001 tMCAO + saline group vs. tMCAO + DPSC-EVs group; two-tailed independent Student’s t-test. C The transfection efficiency of mimic- and inhibitor-miR- 877–3p in DPSCs was quantified by qRT-PCR (n=3). D-J Representative WB bands and densitometric quantification of Bclaf1, MDM2, P53 and apoptosis-related proteins in SH-SY5Y cells after OGD/R (n=3). Data was expressed as mean ± SD. *P<0.05, **P<0.01, ***P<0.001 normoxia group vs. OGD/R + in-miR- 877–3p-EVs group, respectively; ##P<0.01, ###P<0.001 and ####P<0.0001 OGD/R + PBS group vs. OGD/R +mi-miR-877–3p group; Oneway ANOVA, Tukey’s post hoc test.

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) assay Apoptotic cells in cortex were identified using a Tunel assay with an apoptosis detection kit (Proteintech, China), following the manufacturer’s instructions to detect nuclear fragmentation.

Techniques: Inhibition, Saline, Two Tailed Test, Transfection, Quantitative RT-PCR

Journal: Pharmacological research

Article Title: Transfer of miR-877-3p via extracellular vesicles derived from dental pulp stem cells attenuates neuronal apoptosis and facilitates early neurological functional recovery after cerebral ischemia-reperfusion injury through the Bclaf1/p53 signaling pathway.

doi: 10.1016/j.phrs.2024.107266

Figure Lengend Snippet: Fig. 7. Bclaf1 overexpression reversed the effects of DPSC-EVs on apoptosis inhibition. A Transfection efficiency of the LV was detected by fluorescence microscopy, Scale bars in A represent 100 μm (left), 50 μm (upper right) and 20 μm (lower right). B-H Representative WB bands and densitometric quantification of Bclaf1, MDM2, P53 and apoptosis-related proteins in SH-SY5Y cells after OGD/R (n=3). Data was expressed as mean ± SD. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 OGD/R + vector group vs. OGD/R + vector + DPSC-EVs group, respectively; #P<0.05 OGD/R + LV-Bclaf1 vs. OGD/R + LV-Bclaf1 + DPSC-EVs group; Oneway ANOVA, Tukey’s post hoc test. I-J Representative micrographs and quantitative analysis of Tunel-positive cells after OGD/R (n=3). All data was represented as mean ± SD. **P<0.01 OGD/R + vector group vs. OGD/R + vector + DPSC-EVs group, Scale bars in I represent 100 μm.

Article Snippet: Terminal deoxynucleotidyl transferase dUTP nick end labeling (Tunel) assay Apoptotic cells in cortex were identified using a Tunel assay with an apoptosis detection kit (Proteintech, China), following the manufacturer’s instructions to detect nuclear fragmentation.

Techniques: Over Expression, Inhibition, Transfection, Fluorescence, Microscopy, Plasmid Preparation, TUNEL Assay