牛蒡子苷元对七氟烷麻醉致老龄大鼠认知功能障碍的影响及自噬介导的细胞焦亡在其中的作用  

Effect of arctigenin on cognitive dysfunction induced by sevoflurane anesthesia in aged rats and the role of autophagy-mediated pyroptosis

作　　者：王鹏程 王震 张国庆 代彦文 侯铁柱[2] 李治松 Wang Pengcheng;Wang Zhen;Zhang Guoqing;Dai Yanwen;Hou Tiezhu;Li Zhisong(Department of Anesthesiology,Zhumadian Central Hospital,Zhumadian 463000,China;Department of Anesthesiology and Perioperative Medicine,Henan Provincial People′s Hospital,Zhengzhou 450003,China;Department of Anesthesiology,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]驻马店市中心医院麻醉科,驻马店463000 [2]河南省人民医院麻醉与围术期医学科,郑州450003 [3]郑州大学第二附属医院麻醉科,郑州450014

出　　处：《中华麻醉学杂志》2025年第2期163-169,共7页Chinese Journal of Anesthesiology

基　　金：河南省高等学校重点科研项目(22B320116);河南省医学科技攻关计划项目(SBGJ202103058)。

摘　　要：目的评价牛蒡子苷元(ARG)对七氟烷麻醉致老龄大鼠认知功能障碍的影响及自噬介导的细胞焦亡在其中的作用。方法 SPF级雄性SD大鼠50只, 18~20月龄, 体质量550~600 g, 采用随机数字表法分为5组(n=10):对照组(C组)、七氟烷麻醉组(Sev组)、七氟烷麻醉+ARG组(Sev+ARG组)、七氟烷麻醉+自噬诱导剂雷帕霉素组(Sev+RAPA组)、七氟烷麻醉+ARG+自噬抑制剂3-甲基腺嘌呤组(Sev+ARG+3-MA组)。除C组外, 其余各组大鼠吸入6%七氟烷3 h构建认知功能障碍模型。于麻醉前30 min时, Sev+ARG组腹腔注射ARG 20 mg/kg;Sev+RAPA组腹腔注射雷帕霉素7.5 mg/kg;Sev+ARG+3-MA组腹腔注射ARG 20 mg/kg(采用二甲基亚砜进行溶解), 然后立即腹腔注射3-甲基腺嘌呤1.5 mg/kg;C组和Sev组腹腔注射等容量二甲基亚砜。给药结束后48 h时, 行水迷宫实验检测大鼠认知功能。水迷宫实验结束后, 深麻醉下取海马组织, HE染色后观察病理学结果, 并进行海马损伤评分;碘化丙啶染色后检测细胞焦亡情况;采用Western blot法检测神经元自噬相关蛋白[微管相关蛋白轻链3(LC3)、Beclin-1和p62]和焦亡相关蛋白[NOD样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白酶-1前体(pro-caspase-1)、半胱氨酸天冬氨酸蛋白酶-1剪切体(cleaved-caspase-1)、消皮素D(GSDMD)、消皮素D N端片段(GSDMD-N)、白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)]表达。结果与C组相比, Sev组逃避潜伏期延长, 平台象限停留时间缩短, 穿越原平台位置次数减少, 海马损伤评分和细胞焦亡率升高, LC3Ⅱ/LC3Ⅰ比值降低, Beclin-1表达下调, p62、NLRP3、ASC、IL-1β和IL-18表达上调, cleaved-caspase-1/pro-caspase-1比值和GSDMD-N/GSDMD比值升高(P<0.05)。与Sev组相比, Sev+ARG组和Sev+RAPA组逃避潜伏期缩短, 平台象限停留时间延长, 穿越原平台位置次数增加, 海马损伤评分和细胞焦亡率降低, LC3Ⅱ/LC3Ⅰ比值升高, Beclin-1表Objective To evaluate the effect of arctigenin(ARG)on cognitive dysfunction induced by sevoflurane anesthesia in aged rats and the role of autophagy-mediated pyroptosis.MethodsFifty SPF male Sprague-Dawley rats,aged 18-20 months,weighing 550-600 g,were divided into 5 groups(n=10 each)using a random number table method:control group(C group),sevoflurane anesthesia group(Sev group),sevoflurane anesthesia+ARG group(Sev+ARG group),sevoflurane anesthesia+autophagy inducer rapamycin group(Sev+RAPA group),and sevoflurane anesthesia+ARG+autophagy inhibitor 3-methyladenine(3-MA)group(Sev+ARG+3-MA group).Except for group C,the rats in the other groups inhaled 6%sevoflurane for 3 h to establish the cognitive impairment model.At 30 min before anesthesia,ARG 20 mg/kg was intraperitoneally injected in Sev+ARG group,rapamycin 7.5 mg/kg was intraperitoneally injected in Sev+RAPA group,and ARG 20 mg/kg(dissolved in dimethyl sulfoxide)was intraperitoneally injected,followed by immediate intraperitoneal injection of 3-MA 1.5 mg/kg in Sev+ARG+3-MA group.The equal volume of dimethyl sulfoxide was intraperitoneally injected in C group and Sev group.The Morris water maze test was conducted to assess the cognitive function at 48 h after the end of administration.After completion of the Morris water maze test,the hippocampal tissue was taken under deep anesthesia for observation of the pathological changes(after HE staining)which were scored and for determination of neuronal pyroptosis(after propidium iodide staining)and expression of neuronal autophagy-related proteins(microtubule-associated protein 1 light chain 3[LC3],Beclin-1,p62),pyroptosis-related proteins(NOD-like receptor protein 3[NLRP3],apoptosis-associated speck-like protein containing a CARD[ASC],pro-cysteine aspartate-specific protease 1[pro-caspase-1],cleaved-caspase-1,gasdermin D[GSDMD],and N-terminal fragment of gasdermin D[GSDMD-N],interleukin-1β[IL-1β]and IL-18).ResultsCompared with C group,the escape latency was significantly prolonged,the time of staying at the orig

关 键 词：牛蒡子苷 七氟醚 认知功能障碍 老年人 自噬 细胞焦亡 

分 类 号：R614[医药卫生—麻醉学]