摘要
目的探讨丹参酮ⅡA对细菌性脑膜炎(BM)引发海马损伤的保护作用。方法健康、清洁级的SD雄性大鼠40只,采用环磷酰胺(45 mg/kg)和地塞米松(90 mg/kg)腹腔联合注射构建免疫受损动物模型,将受损动物分为空白对照组、鲍曼不动杆菌感染组、生理盐水干预组(菌+生理盐水干预)和丹参酮ⅡA干预组(菌+丹参酮ⅡA干预)4组,构建细菌性脑膜炎的模型;建模第2天开始连续3 d采用水迷宫检测大鼠学习与记忆能力变化,持续感染7 d后处死动物;采用尼氏染色检测海马区神经元细胞的改变,ELISA检测组织内白细胞介素6(IL-6)、白细胞介素10(IL-10)、肿瘤坏死因子β(TNF-β)的浓度,Western blot及RT-PCR检测高迁移率族蛋白1(HMGB1)、基质金属蛋白酶9(MMP-9)、磷酸化的核因子кB(NF-кB/p-p50/p-p65)的蛋白与基因含量变化,评估丹参酮ⅡA对细菌性脑膜炎引发的海马损伤的保护作用;体外以200μmol/L的吡咯烷二硫代氨基甲酸盐(PDTC)抑制神经细胞内NF-кB信号通路的激活,验证体内实验的推论。结果细菌性脑膜炎建模成功后,空白对照和丹参酮ⅡA干预两组大鼠在水迷宫检测的定航与空间探索时限均显著低于细菌性脑膜炎与生理盐水干预两组(P<0.05);尼氏染色显示,丹参酮ⅡA干预后的细菌性脑膜炎组海马区椎体层神经元细胞分布更趋均匀、整齐,尼氏小体数量增多;ELISA检测发现,空白对照和丹参酮ⅡA干预两组血清中IL-6、IL-10、TNF-β浓度显著低于细菌性脑膜炎与生理盐水干预两组(P<0.05);Western blot及RT-PCR检测HMGB1、MMP-9、NF-кB(p-p50,p-p65)蛋白与基因均高表达于细菌性脑膜炎与生理盐水干预两组(P<0.05);体外实验证实在抑制NF-кB通路后丹参酮ⅡA对细菌性脑膜炎感染引发损伤的保护作用并不明显。结论丹参酮ⅡA对细菌性脑膜炎大鼠海马损伤有保护作用,其机制可能与抑制NF-кB通路有关。
Objective To explore the effect and mechanism of tanshinoneⅡA on hippocampal injury in rats with bacterial meningitis(BM).Methods Forty healthy-clean grade SD male rats were intraperitoneally injected with cyclophosphamide(45 mg/kg)and dexamethasone(90 mg/kg)to establish an immune compromised rat model.These rats were divided into four groups:blank control group,BM group(Acinetobacter baumannii infection),BM-NS group(Acinetobacter baumannii infection+normal saline intervention)and BM-TS group(Acinetobacter baumannii infection+tanshinoneⅡA intervention).On the 2 nd day of modeling,rats were examined for changes in their learning and memory abilities using a Morris water maze for 3 consecutive days.After 7 consecutive days of infection,the rats were euthanized.Nissl staining was used to detect the alterations of hippocampal neurons.ELISA was applied to detect the concentrations of interleukin-6(IL-6),IL-10,and tumor necrosis factor-β(TNF-β).Western blot and RT-PCR were used to detect the protein and mRNA expression levels of high mobility group protein 1(HMGB1),matrix metalloproteinase-9(MMP-9)and phosphorylated nuclear factor-кB subunits(NF-кB/p-p50/p-p65).The protective effect of tanshinoneⅡA on BM-induced hippocampal damage was evaluated.To verify the observations in vivo,neural cells were treated with 200μmol/L pyrrolidine dithiocarbamate(PDTC)to inhibit NF-кB signaling pathway.Results After BM model was successful,the navigation and spatial exploration time limits of rats were significantly lower in blank control and BM-TS groups than those in BM and BM-NS groups(P<0.05).Nissl staining showed that the distribution of neuronal cells in the vertebral layer of the hippocampus in BM-TS group was more uniform and orderly,and the number of Nissl bodies was increased after tanshinone IIA intervention.ELISA revealed that the concentrations of serum IL-6,IL-10,and TNF-βwere significantly lower in blank control and BM-TS groups than those in BM and BM-NS groups(P<0.05).Western blot and RT-PCR showed that th
作者
李玉美
余资江
罗时鹏
杨丹
肖朝伦
孙宝飞
黄涛
李琳
刘来兵
LI Yumei;YU Zijiang;LUO Shipeng;YANG Dan;XIAO Chaolun;SUN Baofei;HUANG Tao;LI Ling;LIU Laibing(Department of Anatomy,School of Basic Medical Sciences,Guizhou Medical University,Guiyang 550025,Guizhou,China;Department of Neurosurgery,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)
出处
《贵州医科大学学报》
CAS
2023年第11期1343-1349,1356,共8页
Journal of Guizhou Medical University
基金
贵州省科技厅项目(黔科合基础-ZK〔2023〕-一般313)
贵州省中医药管理局项目(QZYY-2020-054)
贵州省卫生健康委科学技术基金项目(gzwkj2021-522,gzwkj2023-155)
贵州医科大学高层次人才启动基金项目(校博合J字〔2021〕15)。
关键词
细菌性脑膜炎
鲍曼不动杆菌
丹参酮ⅡA
海马损伤
NF-кB通路
PDTC
bacterial meningitis
Acinetobacter baumannii
tanshinoneⅡA
hippocampal damage
NF-кB pathway
pyrrolidine dithiocarbamate
