Fig. 7

Pioglitazone regulates Drp1 phosphorylation, protein oxidation, DNA fragmentation, and neuronal apoptosis in a PPARγ-dependant pathway after TGI. The chemical compounds microinjected into bilateral CA1 subfields as following with DMSO, pioglitazone (20 nmol) 30 min before TGI, or GW9663 (500 ng) 30 min before pioglitazone and 60 min before TGI. Total proteins were isolated from the hippocampal CA1 subfield of sham-operated controls or treated animals 24 h after 10 min of TGI for detection of p-Drp1 (Ser616) in (a) and protein oxidation in (b) and activated caspase-3 expression in (c). DNA was isolated from collected hippocampal CA1 subfield of sham-operated controls, DMSO + I/R, pioglitazone + I/R and GW9662 + pioglitazone 48 h after TGI for detection of DNA fragmentation by PCR assay (d) and hippocampal CA1 tissues were collected 48 h after TGI for detection of DNA fragmentation by sandwich ELISA in (e). Hippocampal slices were subjected to TUNEL staining to determine the extents of apoptosis in (f) which showed sham control in (a), ischemia/reperfusion with vehicle control in (b), pioglitazone with ischemia/reperfusion in (c) and GW9662 + pioglitazone and ischemia/reperfusion in (d). Values are mean ± SEM from representative blots and quantitative analyses from 5–6 animals in each experimental group (a, b and c); values in (e) are fold changes with reference to sham-control; mean ± SEM of 5-6 animals in each experimental group. *P < 0.05 vs. sham-control group, #P < 0.05 vs. DMSO + I/R and + P < 0.05 versus Piog + I/R group in the Scheff′e multiple-range test. I/R: ischemia/reperfusion. Piog: pioglitazone