Fig. 2

The effects of SAM and NaHS administration on neurological deficits and brain edema at 1 and 3 days after ICH. a The schematic diagram shows the four areas (black squares) for Fluoro-Jade C- and TUNEL-positive cell counting in the perihaematomal region. b Representative Fluoro-Jade C staining images and quantitative analyses of Fluoro-Jade C-positive neurons surrounding the haematoma in the sham, vehicle, SAM and NaHS groups at 1 day after ICH. c Representative images of TUNEL-stained (green) and DAPI-stained (blue) brain sections in the perihaematomal area in the sham, vehicle, SAM and NaHS groups at 1 day following operation. d, e Quantitative analyses of Fluoro-Jade C-positive neurons and TUNEL-positive cells surrounding the haematoma in the sham, vehicle, SAM and NaHS groups at 1 day after ICH. Fluoro-Jade C staining demonstrated significantly reduced perihaematomal neuronal cell injury as a result of SAM or NaHS administration at 1 day after ICH (p < 0.05 vs. vehicle). Compared with the sham group, the vehicle group exhibited an increased number of TUNEL-positive cells in the perihaematomal area (p < 0.01). Remarkably, both SAM treatment and NaHS treatment induced a significant reduction in the number TUNEL-positive cells in the perihaematomal area (p < 0.05 vs. vehicle). f Modified Neurological Severity Scores in the sham, vehicle, SAM and NaHS groups at 1 and 3 days after ICH. SAM and NaHS administration significantly revered the neurological deficits assessed at 1 and 3 days after ICH (p < 0.01 vs. sham) at both time points (p < 0.05 vs. vehicle). g, h Brain water content assessment in the sham, vehicle, SAM and NaHS groups at 1 and 3 days after operation. At 1 day after ICH, brain water content was significantly increased in the ipsilateral basal ganglia (p < 0.01) and ipsilateral cortex (p < 0.05) of the ICH-treated groups compared with that in the sham group. SAM or NaHS treatment significantly reduced brain water content in the ipsilateral basal ganglia (p < 0.05 vs. vehicle), but not in the contralateral basal ganglia and ipsilateral cortex (p > 0.05 vs. vehicle). Moreover, there were no significant differences in brain water content in the contralateral basal ganglia and ipsilateral cortex in the ICH-treated groups compared to the vehicle group (p > 0.05). At 3 days after ICH, brain water content was increased only in the ipsilateral basal ganglia after ICH (p < 0.01 vs. sham) and was significantly reduced by both SAM administration and NaHS administration (p < 0.05 vs. vehicle). N = 6 for mNSS and brain water content assessment; n = 5 for Fluoro-Jade C staining; n = 4 for TUNEL staining. Scale bar = 50 μm. Data are presented as the mean ± SEM. *p < 0.05 vs. sham; **p < 0.01 vs. sham; #p < 0.05 vs. vehicle. SAM S-adenosyl-l-methionine, mNSS Modified Neurological Severity Score, ICH intracerebral haemorrhage, TUNEL terminal deoxynucleotidyl transferase dUTP nick end-labelling, DAPI 4′,6-diamidino-2-phenylindole, Ipsi-CX ipsilateral cortex, Cont-CX contralateral cortex, Ipsi-BG ipsilateral basal ganglia, Cont-BG contralateral basal ganglia, Cerebel cerebellum