摘要
目的从环磷酸鸟苷-腺苷合成酶(cGAS)/膜蛋白干扰素基因刺激因子(STING)通路方面,探讨奥拉西坦(ORC)对脑出血大鼠认知功能的影响。方法将SPF级SD大鼠随机分为假手术组、模型组、ORC组(50 mg/kg)、RU.521组(cGAS抑制剂,50 mg/kg)、DMXAA组(STING激活剂,25 mg/kg)、ORC+DMXAA组,每组15只。用Longa法测大鼠神经功能变化;干湿比重法测脑含水量;电镜测血肿周围病理变化;铁染色法测血肿周围组织铁沉积,以评估血肿程度;TUNEL法测血肿周围组织神经元凋亡状况;免疫组化法测血肿周围组织cGAS阳性表达。Western blotting法测STING、TNF-α、IL-12、半胱氨酸天冬氨酸蛋白水解酶3(caspase3)、B淋巴细胞瘤-2基因(Bcl-2)、转铁蛋白(Tf)、转铁蛋白受体(Tf R)、水通道蛋白4(APQ4)蛋白表达。结果与假手术组相比,模型组大鼠死亡、偏瘫及侧旋转等神经功能缺损行为评分明显升高,血肿周围组织神经元水肿及凋亡明显增加,脑水肿、铁沉积严重,cGAS/STING通路蛋白及其介导的炎症反应和促凋亡途径激活(均P<0.05)。ORC组及RU.521组大鼠神经功能缺损评分明显降低、血肿周围组织神经元损伤、凋亡、脑水肿及铁沉积均明显缓解,cGAS/STING通路蛋白及其介导的炎症反应和促凋亡相关蛋白表达明显降低(均P<0.05)。DMXAA可逆转ORC的上述作用(P<0.05)。结论ORC可能通过抑制cGAS/STING通路活化,缓解脑出血大鼠血肿周围神经元炎症损伤及凋亡,减轻神经功能缺损。
Objective From the aspect of cyclic guanosine phosphate-adenosine synthetase(cGAS)/stimulator of interferon gene(STING)pathway,to explore the effect of Oxiracetam(ORC)on the cognitive function of rats with cerebral hemorrhage.Methods SPF SD rats were randomly divided into sham operation group,model group,ORC group(50 mg/kg),RU.521 group(cGAS inhibitor,50 mg/kg),DMXAA group(STING activator,25 mg/kg),ORC+DMXAA group,with 15 rats in each group.Longa method was used to measure changes in rat nerve function;dry and wet specific gravity method was used to measure brain water content;electron microscopy was used to measure the pathological changes around the hematoma;the iron staining method was used to measure the iron deposits in the tissues around the hematoma to assess the degree of the hematoma;TUNEL method was used to measure the neuronal apoptosis in the tissue around the hematoma;immunohistochemical method was used to detect the positive expression of cGAS in tissues around the hematoma.Western blotting was used to detect the expression of STING,TNF-α,IL-12,cysteine aspartate proteolytic enzyme 3(caspase3),B-cell lymphocyte-2(Bcl-2),transferrin(Tf),transferrin receptor(Tf R),aquaporin 4(APQ4)proteins.Results Compared with the sham operation group,the rats in the model group had higher death and neurological dysfunction such as hemiplegia and lateral rotation,increased neuronal edema and apoptosis in the tissue around the hematoma,severe cerebral edema and iron deposition,and activated cGAS/STING pathway protein and their-mediated inflammatory response and pro-apoptotic pathways(all P<0.05).In ORC group and RU.521 group,the scores of neurological dysfunction were significantly reduced,and neuronal damage,apoptosis,cerebral edema and iron deposition in the tissues around the hematoma were significantly alleviated,the expression of cGAS/STING pathway protein and their-mediated inflammatory response and pro-apoptosis related proteins significantly decreased(all P<0.05).DMXAA could reverse the above effects of ORC(P<0.05).Conclusion ORC may inhibit the activation of c GAS/STING pathway to alleviate the inflammatory damage and apoptosis of neurons around hematoma in rats with cerebral hemorrhage and reduce neurological dysfunction.
作者
张涛
谢红兵
高安举
杨清成
ZHANG Tao;XIE Hong-bing;GAO An-ju(Department of Neurosurgery,Anyang People’s Hospital,Anyang 455000,China)
出处
《临床神经病学杂志》
CAS
2022年第3期201-207,共7页
Journal of Clinical Neurology
基金
2019年度河南省医学科技攻关计划联合共建项目(LHGJ20191266)。