Protective Effect Of Hydrogen Sulfide On Brain Damage For Experimental Subarachnoid Hemorrhage

Objectives:Hydrogen sulfide (H2S) has a protective effect on the brain injury, but theprotective effect on subarachnoid hemorrhage (SAH) brain damage has not beenreported. The aim of the present study was to investigate the new role of H2S treatmentfor SAH by detecting the expression of GFAP、MMP-9、c-fos、caspase-3、Bcl-2.Methods:1. Estabished a subarachnoid hemorrhage animal model according to Ayer RE andto be improved.2. Experimental groups: Rats were divided into3experimental groups:（1）shamgroup, Intraperitoneal injection of0.5mL saline；（2）SAH model group, as modelprogram and intraperitoneal injection of0.5mL saline；（3）H2S treatment group, aftera model program, intraperitoneal injection of0.5mL/0μmol/kg,6.25μmol/kg,12.5μmol/kg,25μmol/kg NaHS respectively.3. The experimental rats of the SAH model were injected NaHS50mg/Kg bodyweight in intraperitoneal and after surviving for24hours to take the whole brain. Theexpression changes of GFAP, Bcl-2, c-fos, caspase-3, MMP-9were detected withfluorescent immunocytochemistry and Western blot method.4. Statistical analysis: SPSS13.0software package was used for statistical analysiseach set of data, one-way ANOVA was used to compare between groups,0.05wasconsidered statistically significant, the results of Western blot were analysised byImagej and GraphPad Prism5software, the value represented by the light density. Results:1. Protective effects of H2S on early brain injury SAH: neurological score wasobserved after estabished a subarachnoid hemorrhage animal model, the results showH2S paly a significant role in improving brain function, the mortality rate from50%to10%.2. Effects of hydrogen sulfide on the expression of GFAP: the expression ofGFAP was detected by immunofluorescence and Western-blot on hippocampal cortexand frontal and parietal lobe, The results show sham group astrocyte GFAP expressionwas low, SAH model group was significantly increased (P <0.05), Compared with themodel group, the expression was significantly reduced hydrogen sulfide group (P<0.01).3. Effects of hydrogen sulfide on the expression of c-fos and MMP-9:Western-blot detection show expression of MMP-9and c-fos protein was significantlyincreased SAH model group compared with the sham group (P <0.01), and decreasedexpression of hydrogen sulfide group (P <0.05).4. Effects of hydrogen sulfide on the expression of Bcl-2and caspase-3:immunofluorescence and Western-blot detection show hippocampal cortex of Bcl-2positive cells were mainly distributed in the C2and C3zones, Bcl-2positive cells wastested in the sham and SAH group, but Bcl-2positive cells was significantly increasedin the hydrogen sulfide group (P<0.05); immunofluorescence and Western-blotdetection show SAH model group increased expression of caspase-3positive cellsincreased significantly compared with sham group, while the hydrogen sulfide groupcaspase-3positive cells was significantly reduced (P <0.001).Conclusion:1、 The expression of GFAP was increased on hippocampal cortex and frontaland parietal lobe after SAH. GFAP expression was significantly reduced after H2Streatment，H2S protective effect on brain injury may be associated with improvement inglial cell function. 2、 The expression of MMP-9and c-fos was increased on hippocampal cortexand frontal and parietal lobe after SAH, both factors was significantly reduced by H2Sintervention, this suggests that H2S has a protective effect to neurons.3、 the expression of apoptotic factors caspase-3and Bcl-2have significantlyimproved after H2S treatment, it showed protective effects of H2S on brain injuryrelated to the mechanism of apoptosis.