硫化氢对蛛网膜下腔出血大鼠海马区神经损伤的保护作用及机制研究

Effect of hydrogen sulfide on brain damage and its mechanism in rats with subarachnoid hemorrhage

目的 探讨硫化氢对蛛网膜下腔出血（SAH）大鼠海马区神经损伤的影响及其作用机制. 方法 30只清洁级SD大鼠按随机数字表法分成3组：对照组、SAH模型组、硫化氢组,每组各10只.后2组大鼠应用大脑中动脉穿刺法制作成SAH模型,硫化氢组于造模后腹腔注射100mg/kg硫氢化钠.各组大鼠于造模后24h进行神经功能评分,后取海马组织切片,采用荧光免疫组化染色和Western blotting等方法检测海马区神经细胞胶质纤维酸性蛋白（GFAP）、S-100B、Bcl-2、caspase-3、c-fos、基质金属蛋白酶-9（MMP-9）蛋白的表达变化. 结果 硫化氢组神经功能评分（2.1±0.8）较SAH模型组（3.1±0.7）相比明显降低,差异有统计学意义（P＜0.05）.荧光免疫组化染色结果显示：SAH模型组GFAP、S-100B蛋白表达较对照组明显增高,差异有统计学意义（P＜0.05）;而应用硫化氢干预后二者表达较SAH模型组明显下降,差异均有统计学意义（P＜0.05）.SAH模型组Bcl-2、caspase-3均有表达,而应用硫化氢干预后Bcl-2表达较SAH模型组明显增多、caspase-3表达明显下降,差异均有统计学意义（P＜0.05）.Western blotting检测结果显示：SAH模型组c-fos、MMP-9表达较对照组均明显增高,差异有统计学意义（P＜0.05）;而应用硫化氢干预后二者表达与SAH模型组相比明显下降,差异均有统计学意义（P＜0.05）. 结论 硫化氢对SAH后脑损伤有明显保护作用,其机制可能与改善胶质细胞的功能及相关凋亡机制有关.

Objective To explore the effect of hydrogen sulfide （H2S） on brain injury in rats with subarachnoid hemorrhage （SAH） and its mechanism.Methods Thirty SD rats were randomly divided into control group （n=10）,SAH model group （n=10） and NaHS treatment group （n=10）; middle cerebral artery puncture was performed in rats in the later two groups,and rats in the NaHS （100 mg/kg）treatment group were given NaHS 100 mg/kg via intraperitoneal injection.Twenty-four h after the puncture,the rats were sacrificed; hippocampus slices were prepared; the expression changes ofglial fibrillary acidic protein （GFAP）,S-100B,Bcl-2,C-fos,caspase-3 and matrix metalloproteinase-9 （MMP-9） were detected by fluorescent immtmocytochemistry and Western blotting.Results Neurological assessment showed that severe neurological symptoms in the NaHS treatment group （2.1±0.8 points） were significantly relieved as compared with those in SAH model group （3.1 ±0.7 points,P〈0.05）.GFAP and S-100B in SAH model group enjoyed higher expressions than those in the control group,while those in the NaHS treatment group were significantly lower than those in the SAH model group （P〈0.05）.The expressions of Bcl-2 and caspase-3 in SAH group were higher than those in the control group,but the expression of Bcl-2 was significantly increased and caspase-3 was statistically decreased in NaHS treatment group than that in the SAH model group （P〈0.05）.Western blotting showed that the expressions of c-fos and MMP-9 in SAH model group were increased as compared with those in the control group,but those in the NaHS treatment group were significantly decreased as compared with those in the SAH model group （P〈0.05）.Conclusion Hydrogen sulfide has a significant protective effect on brain injury after SAH and its mechanism may be related to the function of glial cells and apoptotic pathways.