Journal: Chinese Medicine
Article Title: Thonningianin A derived from Penthorum chinense Pursh alleviates cerebral ischemia/reperfusion-mediated apoptosis and pyroptosis through the activation of PINK1/Parkin-dependent mitophagy
doi: 10.1186/s13020-025-01247-2
Figure Lengend Snippet: TA improved mitochondrial oxidative damage. A The death rate of HT22 cells was measured by Hoechst/PI staining in HT22 cells after 24 h of OGD/R induction and TA treatment. Magnification: 10 × , scale bar: 200 μm. B Representative images showing ROS release (DHE staining) and viable cells (Hoechst staining) in HT22 cells following OGD/R with or without TA treatment, × 10, scale bar: 200 μm. Bar chart indicates the rate of PI/Hoechst. *** p < 0.001 versus OGD/R alone group, n = 3. C Representative JC-1 staining images depict MMP in HT22 cells following OGD/R with or without TA treatment. Red fluorescence indicates JC-1 aggregates, while green fluorescence indicates the monomeric form. Magnification: 10 × , scale bar: 200 μm. D HT22 cells underwent OGD/R and TA treatment, followed by incubation with Hoechst reagent and Mito-Tracker ™ Red CMXRos to stain nuclei and mitochondria. Images were acquired using a confocal laser scanning microscope. Magnification: 64 ×, scale bar: 5 μm. E The bar chart indicates the DHE/Hoechst rate of HT22 cells. * p <0.05 and *** p < 0.001 versus OGD/R alone group, n = 3. F After 24 h of OGD/R induction and TA treatment, HT22 cell viability was measured by MTT assay, the result is as shown in the bar chart. * p < 0.05, ** p < 0.01 and *** p < 0.001 , and versus OGD/R group, n = 6. G The bar chart indicates the DHE/Hoechst rate of HT22 cells. *** p < 0.001 versus OGD/R alone group, n = 3. H The bar chart illustrates the ratio of JC-1 aggregates to monomers in HT22 cells. *** p < 0.001 versus OGD/R alone group, n = 3. I The bar chart indicates mitochondrial length of HT22 cells in different group. *** p < 0.001 versus OGD/R alone group, n = 3. J Detection of nuclear Nrf2, HO-1, NQO1, and GCLC antioxidant protein expression by Western blotting. K HT22 cells subjected to OGD/R with or without TA, or TA + ML385 (an Nrf2 inhibitor), Western blotting was used to detect Nrf2 protein expression levels. L – O The bar chart indicates the ratios of nuclear Nrf2/LaminB, HO-1/β-actin, NQO1/GAPDH, GCLC/GAPDH. * p < 0.05, ** p < 0.01 and *** p < 0.001 versus OGD/R alone group. P The bar chart demonstrates that TA treatment enhanced OGD/R-induced nuclear Nrf2 expression, an effect reversed by the Nrf2 inhibitor ML385. *** p < 0.001 versus OGD/R group or OGD/R + TA group
Article Snippet: Antibodies against GSDMD (20770-1-Ap), Caspase-3 (25158-1-Ap), Caspase-9 (10380-1-Ap), NQO1 (11451-1-Ap), GCLC (12601-1-Ap), LC3 (14600-1-Ap), PINK1 (23274-1-Ap), Parkin (14060-1-Ap), Antibody Caspase-1 (22,915-1-ap), GAPDH (6004-1-Ig), β-actin (66009-1-Ig), CoraLite ® Plus 488 goat anti-rabbit IgG (RGAR002), CoraLite ® Plus 594-Goat Anti-Mouse IgG (RGAR002), were purchased from Proteintech Group, Inc (Wuhan, China).
Techniques: Staining, Fluorescence, Incubation, Laser-Scanning Microscopy, MTT Assay, Expressing, Western Blot
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