目的 研究胆南星Arisaema Cum Bile多糖组分拮抗高热惊厥模型大鼠神经炎症作用的可能机制。方法 采用水提醇沉法制备胆南星多糖，并通过色谱、光谱等方法对其进行结构表征分析；通过ip脂多糖联合热水浴建立高热惊厥模型，随机分为对照组、模型组、丙戊酸钠（200 mg/kg）组和胆南星高、低剂量（200、100 mg/kg）组，每组10只。记录每组大鼠惊厥潜伏期，惊厥持续时间和惊厥级别，并于末次造模2 h后测量大鼠肛温；苏木素-伊红（hematoxylin eosin，HE）染色评估海马组织神经元病理变化；ELISA法检测血清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、IL-6及海马组织中γ-氨基丁酸（γ-aminobutyricacid，GABA）、谷氨酸（glutamic acid，Glu）、NOD样受体热蛋白结构域3（NOD like receptor family pyrin domain containing 3，NLRP3）和高迁移率族蛋白B1（high mobility group box 1 protein，HMGB1）水平；采用超高效液相色谱-四级杆飞行时间质谱（ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry，UPLC-Q-TOF-MS/MS）对大鼠的血清进行代谢组学分析。结果 分离制备得到的胆南星多糖中总糖、蛋白质和糖醛酸质量分数分别为79.4%、2.96%和2.35%；气相色谱-质谱联用技术（gas chromatography-mass spectrometry，GC-MS）测定胆南星多糖主要由阿拉伯糖、木糖、甘露糖、葡萄糖和半乳糖组成，其物质的量比分别为0.030∶0.260∶0.115∶0.741∶0.088；傅里叶变换红外光谱仪（Fouriertransform infrared spectrometer，FT-IR）光谱表明多糖含有吡喃环；与模型组比较，胆南星多糖高剂量组大鼠肛温降低（P＜0.01）且胆南星多糖高、低剂量组均可显著延长惊厥潜伏期（P＜0.01），缩短惊厥持续时间（P＜0.01）；对海马神经元细胞具有保护作用，升高海马组织GABA含量，同时降低血清中TNF-α、IL-1β和IL-6与海马组织中Glu、NLRP3及HMGB1含量（P＜0.05、0.01）；代谢组学结果表明，胆南星多糖可显著回调16种生物标志物的表达，共涉及花生四烯酸代谢、氨基酸代谢及嘧啶代谢等6条代谢通路径。结论 胆南星多糖组分对高热惊厥大鼠表现出显著的拮抗作用，其作用机制可能与解热、保护海马神经元、抑制脑内炎症发生及调节代谢紊乱有关。

Objective To investigate the possible mechanisms by which polysaccharides of Dannanxing (Arisaema Cum Bile) inhibit neuroinflammation in febrile seizure rat model. Methods The polysaccharides of Arisaema Cum Bile were prepared by a combination of water extraction and alcohol precipitation, and its primary structure was characterized by chromatographic and spectral methods. Febrile seizure model was established by intraperitoneal injection of lipopolysaccharide combined with hot water bath. The rats were randomly divided into five groups, including control group, model group, sodium valproate group (200 mg/kg), Arisaema cum bile polysaccharides high- and low-dose (200, 100 mg/kg) groups, with ten rats in each group. The convulsive latency, duration and grade of convulsion were recorded in each group, and the rectal temperature of rats was monitored 2 h after the last modeling. Hematoxylin eosin (HE) staining was used to evaluate the pathological changes of hippocampal neurons. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6 in serum and γ-aminobutyricacid (GABA), glutamic acid (Glu), NOD like receptor family pyrin domain containing 3 (NLRP3) and high mobility group box 1 protein (HMGB1) in hippocampus tissues were detected by ELISA. The metabolomics analysis of rat serum based on ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) was performed. Results The total sugar, protein and glucuronic acid contents of the Arisaema Cum Bile polysaccharides obtained from the isolation preparation were 79.4%, 2.96% and 2.35%, respectively. The results of monosaccharide composition based on gas chromatography-mass spectrometry (GC-MS) exhibited that the polysaccharides of Arisaema Cum Bile were mainly composed of arabinose, xylose, mannose, glucose and galactose with molar mass ratios of 0.030∶0.260∶0.115∶0.741∶0.088, respectively. Fouriertransform infrared spectrometer (FT-IR) spectroscopy indicated that the polysaccharides contained pyran ring. Compared with the model group, the rectal temperature of rats in the high-dose group of Arisaema Cum Bile polysaccharide was decreased (P < 0.01), and both high and low dose groups of Arisaema Cum Bile polysaccharide significantly prolonged the convulsion latency (P < 0.01) and shortened the duration of convulsions (P < 0.01). The high-dose Arisaema cum bile polysaccharides supplementation showed a protective effect on hippocampal neuronal cells, elevated GABA content in hippocampal tissues, and simultaneously reduced the content of TNF-α, IL-1β, IL-6 in serum, Glu, NLRP3 and HMGB1 in hippocampal tissues (P < 0.05, 0.01). The metabolomics results showed that Arisaema Cum Bile polysaccharides could significantly reverse the expression of 16 biomarkers, which involved six metabolic pathways including arachidonic acid metabolism, amino acid metabolism and pyrimidine metabolism. Conclusion The polysaccharide fraction of Arisaema Cum Bile showed significant antagonistic effects on febrile seizure rats, and its underlying mechanism may be closely related to lowering body temperature, protecting hippocampal neurons, inhibiting neuroinflammation in brain and regulating metabolic disorders.

R285.5

2022年全国名老中医药专家传承工作室建设项目（国中医药人教函[2022]75号）;第七批全国老中医药专家学术经验继承工作项目（国中医药人教函[2022]76号）;黑龙江省“头雁”团队支持项目（黑龙江省头雁行动领导小组文件[2019]5号）