淫羊藿苷对失血性休克复苏模型小鼠认知功能及星形胶质细胞焦亡的影响  

Effects of icariin on cognitive function and astrocytic pyroptosis in hemorrhagic shock resuscitation model mice

作　　者：张立民 宋荣欣 单玉栋 贾世艳 吕金梦 王璐颖 井郁陌[1] Zhang Limin;Song Rongxin;Shan Yudong;Jia Shiyan;Lyu Jinmeng;Wang Luying;Jing Yumo(Department of Anesthesiology,Hebei Province Cangzhou Hospital of Integrated Traditional and Western Medicine,Cangzhou 061000,China;Graduate School,Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]河北省沧州中西医结合医院麻醉科,沧州061000 [2]河北医科大学研究生院,石家庄050011

出　　处：《中华行为医学与脑科学杂志》2023年第2期104-110,共7页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金(82171455);河北省自然科学青年科学基金(H2021110004)。

摘　　要：目的:探讨淫羊藿苷对失血性休克复苏模型小鼠认知功能及星形胶质细胞焦亡的影响。方法:48只SPF级健康雄性C57BL/6小鼠按照随机数字表法分为4组(每组n=12):假手术对照组(C组)、失血性休克复苏组(H组)、失血性休克复苏+淫羊藿苷组(HI组)和失血性休克复苏+淫羊藿苷+SSK1组[HIS组,其中SSK1为p38丝裂原活化的蛋白激酶(p38MAPK)磷酸化激动剂]。H、HI和HIS组小鼠通过股静脉放血回输法建立失血性休克复苏模型;HI和HIS组在复苏第2天行淫羊藿苷(10 mg/kg)灌胃,连续7 d;C组和H组仅向小鼠灌胃含二甲基亚砜的等量0.9%氯化钠溶液。HIS组小鼠在复苏第2天行SSK1(0.5 mg/kg)腹腔注射,连续7 d;C、H和HI组仅向腹腔注射含二甲亚砜的等量0.9%氯化钠溶液。于术后15 d,采用新物体识别实验和恐惧条件实验评价小鼠认知功能。通过免疫荧光染色法测定小鼠海马区神经元特异标记蛋白微管相关蛋白2(microtubule-associated protein 2,MAP2)及焦亡神经胶质细胞特异蛋白并计算海马CA1区神经元含量及星形胶质细胞焦亡率;通过Western blot法检测海马区焦亡相关炎性因子白介素(interleukin,IL)-1β、IL-18、磷酸化p38MAPK和总p38MAPK的表达。采用SPSS 21.0软件进行统计分析,多组间比较采用单因素方差分析,进一步两两比较用SNK-q检验。结果:新物体识别实验结果显示,4组小鼠的新物体识别指数差异有统计学意义(F=50.75,P<0.05)。H组、HI及HIS组小鼠新物体识别指数[(22.7±6.9),(40.1±7.0),(22.5±7.5)]显著低于C组(58.5±11.2),HI组小鼠新物体识别指数高于H组,HIS组小鼠新物体识别指数低于HI组(均P<0.05)。恐惧条件实验显示,4组小鼠僵住时间百分率差异有统计学意义(F=60.54,P<0.05)。H组、HI及HIS组小鼠僵住时间百分率[(21.8±5.0)%,(38.4±7.4)%,(21.3±4.2)%]显著低于C组[(49.1±7.0)%],HI组小鼠僵住时间百分率高于H组,HIS组小鼠僵住时间百分率低于HI组(均P<0.05)�Objective To investigate the icariin on cognitive function and astrocytic pyroptosis in hemorrhagic shock resuscitation model mice.Methods Forty-eight SPF grade C57BL/6 mice(male)were randomly divided into four groups(n=12 in each group):Sham operation control group(Group C),hemorrhagic shock and resuscitation group(Group H),hemorrhagic shock and resuscitation plus icariin group(Group HI)and hemorrhagic shock resuscitation plus icariin and SSK1 group(Group HIS,SSK1 was a phosphorylation agonist of mitogen-activated protein kinase p38(p38MAPK)).The mice in Group H,HI and HIS were subjected to hemorrhagic shock and resuscitation model by bleeding and retransfusion via left femoral vein;the mice in Group HI and HIS were administered with icariin(10 mg/kg)intragastrically for 7 days;the mice in Group C and H were administered with the same amount of normal saline containing dimethyl sulfoxide(DMSO).The mice in Group HIS were administered with SSK1(0.5 mg/kg)intraperitoneally,but the mice in Group C,H and HI were only administered with the same amount of normal saline containing DMSO.At 15 days after resuscitation,novel objective recognition test and fear conditioning test were used to assess cognitive dysfunction of mice.Microtubule-associated protein 2(MAP2),a specific marker protein of neurons reflecting astrocytic pyroptosis in the hippocampus of mice,were detected by immunofluorescence assay so as to assess neuronal injury and astrocytic pyroptosis.The levels of IL-1β,IL-18,the ratio of phosphorylated p38MAPK to total p38MAPK in the hippocampus were evaluated by Western blot.SPSS 21.0 software was used for data analysis,multiple samples among groups were compared by one-way ANOVA,and SNK-q test was used for further pairwise comparison.Results The results of new object recognition test showed that the difference of new object recognition index among the four groups was statistically significant(F=50.75,P<0.05).The new object recognition indexes in H group(22.7±6.9),HI group(40.1±7.0)and HIS group(22.5±7.5)were

关 键 词：失血性休克 淫羊藿苷 焦亡 p38丝裂原激活的蛋白激酶 认知功能障碍 小鼠 

分 类 号：R285.5[医药卫生—中药学]