青蒿琥酯通过抑制NLRP3炎性体激活和炎性细胞因子分泌减轻新生大鼠缺氧缺血性脑损伤  

Artesunate alleviates hypoxic⁃ischemic brain damage in neonatal rats by inhibiting NLRP3 inflammasome activation and inflammatory cytokine secretion

作　　者：曹银利[1] 孙亚洲 崔清洋 何晓敬 李珍珍 CAO Yinli;SUN Yazhou;CUI Qingyang;HE Xiaojing;LI Zhenzhen(Department of Pediatrics,The First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453000,China)

机构地区：[1]新乡医学院第一附属医院儿科,河南新乡453000

出　　处：《细胞与分子免疫学杂志》2023年第5期410-422,共13页Chinese Journal of Cellular and Molecular Immunology

基　　金：河南省医学科技攻关计划(联合共建)项目(LHGJ20190444)。

摘　　要：目的研究青蒿琥酯对新生大鼠缺血缺氧性脑损伤(HIBD)的保护作用及其作用机制。方法7日龄新生SD大鼠随机分为假手术组、模型组、5 mg/kg青蒿琥酯组、10 mg/kg青蒿琥酯组、20 mg/kg青蒿琥酯组、6 mg/kg地塞米松组,每组18只。除假手术组外,其余各组建立HIBD模型,假手术组只需分离左颈侧总动脉,不行结扎和氮氧混合气体通气处理。在手术后,各组立刻腹腔注射对应药物,假手术组和模型组大鼠注射等体积的生理盐水,之后每日一次,共给药5次。末次给药1 h后,对各组大鼠进行神经功能缺损评分,大鼠处死后检测脑含水量,观察大鼠海马CA1区脑组织病理学改变,原位末端转移酶标记技术(TUNEL)检测神经细胞凋亡情况,ELISA分别检测各组大鼠脑组织和外周血中白细胞介素1β(IL⁃1β)、IL⁃6、肿瘤坏死因子α(TNF⁃α)的水平,Western blot法检测各组大鼠海马CA1区脑组织含pyrin结构域核苷酸结合寡聚结构域样受体家族蛋白3(NLRP3)、含胱天蛋白酶募集结构域凋亡相关斑点样蛋白(ASC)、胱天蛋白酶1(caspase⁃1)蛋白表达。结果与模型组比较,(10、20)mg/kg青蒿琥酯组和地塞米松组大鼠神经功能缺损评分下降,脑组织病理损伤减轻,脑含水量明显减少,海马神经元细胞凋亡数明显减少,脑组织和外周血中IL⁃1β、IL⁃6、TNF⁃α水平明显降低,NLRP3、ASC、caspase⁃1水平明显降低。结论青蒿琥酯能够改善HIBD新生大鼠的神经功能、缓解脑部损伤、减轻脑水肿,对HIBD起保护作用,这可能与抑制NLRP3炎性体激活,减少炎性细胞因子的分泌有关。Objective To investigate the protective effect of artesunate on hypoxic⁃ischemic brain damage(HIBD)and its mechanism in neonatal rats.Methods 7⁃day⁃old neonatal SD rats were randomly divided into sham operation group,model group,artesunate 5 mg/kg group,artesunate 10 mg/kg group,artesunate 20 mg/kg group and dexamethasone 6 mg/kg group,with 18 rats in each group.HIBD models were established in groups except for the sham operation group.The sham operation group only needed to separate the left common carotid artery without ligation and nitrogen⁃oxygen mixed gas ventilation.Each group was injected with drug intraperitoneally right after surgery and the rats in the sham operation group and the model group were injected with an equal volume of normal saline(once a day for a total of 5 times).One hour after the last injection,the rats in each group were scored for neurological defects.After the rats were sacrificed,the brain water content was measured and the pathological changes of the brain tissues of rats were observed.Terminal⁃deoxynucleotidyl transferase mediated nick end labeling(TUNEL)was used to detect the neuronal cell apoptosis,and ELISA was applied to detect the levels of IL⁃1β,IL⁃6 and TNF⁃αin brain tissues and peripheral blood of each group of rats.Western blot analysis was adopted to detect the protein expression levels of NLR family pyrin domain containing 3(NLRP3),apoptosis⁃associated speck⁃like protein containing CARD(ASC)and caspase⁃1 in the rats brain tissues of each group.Results Compared with the model group,the neurological deficit score was decreased;the pathological damage of brain tissues was relieved;the brain water content was significantly reduced;the apoptosis number of hippocampal neurons was decreased significantly;the levels of IL⁃1β,IL⁃6 and TNF⁃αin brain tissues and peripheral blood were significantly reduced;the protein expression levels of NLRP3,ASC and caspase⁃1 were significantly lowered in the middle⁃dose and high⁃dose artesunate groups and t

关 键 词：青蒿琥酯 缺氧缺血性脑损伤(HIBD) 含pyrin结构域核苷酸结合寡聚结构域样受体家族蛋白3(NLRP3) 炎性体 神经细胞 凋亡 

分 类 号：R285.5[医药卫生—中药学] R364.4[医药卫生—中医学] R743.31