香叶木素对细菌性脑膜炎模型大鼠海马细胞铁死亡的影响及SIRT1-Nrf2信号通路机制  

Effect of diosmetin on ferroptosis of hippocampal cells in rats with bacterial meningitis and the mechanism of SIRT1-Nrf2 signaling pathway

作　　者：张路 王紫婷 刘熙鹏[2] 张秀峰[2] Zhang Lu;Wang Ziting;Liu Xipeng;Zhang Xiufeng(Graduate School,Hebei North University,Zhangjiakou 075000,China;Department of Neurosurgery,the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院研究生院,张家口075000 [2]河北北方学院附属第一医院神经外科,张家口075000

出　　处：《中华行为医学与脑科学杂志》2023年第7期584-591,共8页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：河北省医学科学研究课题计划项目(20190872)。

摘　　要：目的:探究香叶木素(diosmetin,Dio)对细菌性脑膜炎(bacterial meningitis,BM)大鼠神经元铁死亡的影响及作用机制。方法:选用SPF级6~7周龄雄性SD大鼠,采用大鼠小脑延髓池注射B族溶血性链球菌法建立BM模型,将建模成功的60只BM模型大鼠按照随机数字表法分为模型组、Dio低、中、高剂量组和抑制剂组,每组12只。另取12只体质量相匹配的大鼠作为对照组。Dio低、中、高剂量组和抑制剂组大鼠分别按照50 mg/kg、100 mg/kg、200 mg/kg、200 mg/kg剂量的Dio灌胃,对照组则灌胃等体积0.9%氯化钠溶液,灌胃当天抑制剂组大鼠腹腔注射沉默信息调节因子1(silent mating type information regulation homolog 1,SIRT1)通路抑制剂EX527(10 mg/kg),其余各组大鼠则注射等体积0.9%氯化钠溶液。以上干预1次/d,连续28 d。采用Loeffler神经学评分评估大鼠神经功能损伤情况,ELISA法检测检测大鼠脑脊液中白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),血细胞分析仪检测脑脊液中白细胞数量,采用微量酶标法检测谷胱甘肽(glutathione,GSH)、硫代巴比妥酸显色法检测丙二醛(malondialdehyde,MDA)、比色法测定活性氧(reactive oxygen species,ROS)、亚铁嗪法检测Fe 2+水平,苏木精-伊红染色、普鲁士蓝染色和TUNEL染色分别观察海马组织病理损伤、铁积累及海马区细胞凋亡情况,Western blot检测转铁蛋白(transferrin,Tf)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)、半胱氨酸蛋白酶3(caspase-3)及SIRT1/Nrf2/HO-1/Gpx4信号通路蛋白表达。使用Graphpad Prism 9.0进行数据分析,多组间比较采用单因素方差分析,进一步两两比较采用SNK-q检验。结果:(1)6组大鼠神经功能评分差异有统计学意义(F=125.451,P<0.001)。模型组大鼠神经功能评分低于对照组,Dio低、中、高剂量组大鼠神经功能评分均高于模型组(均P<0.0ObjectiveTo explore the effect and mechanism of diosmetin(Dio)on neuronal ferroptosis in rats with bacterial meningitis(BM).MethodsMale SD rats aged 6-7 weeks of SPF grade were selected for the experiment.The BM model was established by injecting group B hemolytic streptococcus into the cisterna magna of cerebellum.Sixty BM model rats were successfully modeled and divided into model group,low-dose Dio group,medium-dose Dio group,high-dose Dio group and inhibitor group according to the random number table method,with 12 rats in each group.Another 12 weight-matched rats were taken as the control group.The rats in the low-dose Dio group,medium-dose Dio group,high-dose Dio group and the inhibitor group were intragastrically administered with Dio at 50 mg/kg,100 mg/kg,200 mg/kg and 200 mg/kg,respectively.The rats in the control group were intragastrically administered with an equal volume of 0.9%sodium chloride solution.On the day of intragastric administration,the rats in the inhibitor group were intraperitoneally injected with SIRT1 pathway inhibitor EX527(10 mg/kg),and the rats in the other groups were injected with an equal volume of 0.9%sodium chloride solution.The above interventions were performed once a day for 28 consecutive days.Loeffler neurological score was used to evaluate the neurological impairment in rats.Interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in cerebrospinal fluid of rats were detected by ELISA.The number of white blood cells in cerebrospinal fluid was detected by a blood cell analyzer.Glutathione(GSH)was detected by micro-enzyme labeling method,malondialdehyde(MDA)was detected by thiobarbituric acid colorimetric method,reactive oxygen species(ROS)was detected by colorimetry,and Fe2+level was detected by ferrozine method.Hematoxylin-eosin staining,Prussian blue staining and TUNEL staining were used to observe the pathological damage,iron accumulation and apoptosis in the hippocampus,respectively.Western blot was applied to measure the expression of transferrin(Tf),proliferating cell

关 键 词：香叶木素 沉默信息调节因子1 核因子E2相关因子2 血红素加氧酶1 谷胱甘肽过氧化物酶4 细菌性脑膜炎大鼠 铁死亡 

分 类 号：R742.9[医药卫生—神经病学与精神病学]