This work studied the impact of brain peptide(BP)on sleep and intestinal microbiota in mice, and analyzed the relationship between intestinal microbiota and sleep through changes in neurotransmitter content and oxidative stress levels in mice. The insomnia model was established by peritoneal injection of p-chlorophenylalanine(PCPA), which was administered via continuous gavage for 7 days. The sedative and hypnotic effects of brain peptides on insomniac mice were analyzed by using a pentobarbital sodium sleep coordination test, tail suspension test(TST),light and dark box test(LDB), along with measurements of- DOI:10.3969/j.issn.1000-1565.2024.05.005脑多肽对PCPA诱导的睡眠障碍小鼠5-羟色胺系统和肠道菌群的影响黄欣然,王琳娜,代佳琪,宋楠楠,周程艳(河北大学 药学院,河北省药物质量分析控制重点实验室,河北 保定 071002)摘 要:为探究脑多肽对昆明小鼠睡眠和肠道菌群的影响,通过小鼠神经递质含量和氧化应激水平的变化,分析肠道菌群与睡眠之间的联系.腹腔注射对氯苯丙氨酸(PCPA)建立睡眠障碍模型,连续灌胃给药7 d,通过戊巴比妥钠睡眠协同实验、悬尾和明暗箱实验以及小鼠神经递质和氧化应激水平的变化,分析脑多肽对睡眠障碍小鼠镇静催眠功能的影响;苏木精-伊红染色法(H&E)和糖原染色法(AB-PAS)观察小鼠海马及结肠组织病理变化;收集粪便,进行16S rRNA基因测序,分析小鼠肠道菌群丰度的变化.结果表明:脑多肽高、低剂量组小鼠睡眠时间显著增加,睡眠潜伏期显著缩短,MDA含量显著降低,5-HT、GABA、MT含量、GSH-Px和SOD酶活均显著升高(P<0.05);小鼠海马CA1区神经元细胞排列整齐,胞核固缩深染现象改善,结肠组织中炎症细胞浸润减少;脑多肽高剂量组拟杆菌门丰度显著升高,乳杆菌属、双歧杆菌属丰度显著升高(P<0.05),拟杆菌属丰度与GSH-Px的变化趋势呈显著正相关,乳杆菌属丰度与5-HT、MT、SOD的变化趋势呈显著正相关(P<0.05).脑多肽通过调节肠道菌群正向调节血清内神经递质和脑组织氧化应激水平,有效治疗PCPA诱导的小鼠睡眠障碍症状,随着剂量增加治疗效果明显提高.关键词:脑多肽;5-羟色胺;肠道菌群;氧化应激;睡眠障碍中图分类号:R917 文献标志码:A 文章编号:1000-1565(2024)05-0483-12Effects of brain peptides on 5-hydroxytryptamine system and intestinal flora in PCPA induced sleep disorder miceHUANG Xinran, WANG Linna, DAI Jiaqi, SONG Nannan, ZHOU Chengyan(Key Laboratory of Pharmaceutical Quality Control of Hebei Province, College of Pharmacy Sciences, Hebei University, Baoding 071002, China)Abstract: This work studied the impact of brain peptide(BP)on sleep and intestinal microbiota in mice, and analyzed the relationship between intestinal microbiota and sleep through changes in neurotransmitter content and oxidative stress levels in mice. The insomnia model was established by peritoneal injection of p-chlorophenylalanine(PCPA), which was administered via continuous gavage for 7 days. The sedative and hypnotic effects of brain peptides on insomniac mice were analyzed by using a pentobarbital sodium sleep coordination test, tail suspension test(TST),light and dark box test(LDB), along with measurements of- 收稿日期:2024-04-04;修回日期:2024-05-18 基金项目:河北省自然科学基金资助项目(H2020201291);贵州医科大学省部共建药用植物功效与利用国家重点实验室资助项目(FAMP202006K) 第一作者:黄欣然(2003—),女,河北大学在读硕士研究生,主要从事睡眠疾病菌群相关研究以及药物分析方向研究.E-mail:huang18132083386@163.com 通信作者:周程艳(1976—),女,河北大学副教授,主要从事代谢组学、菌群、脂质组学以及相关药理作用机制研究.E-mail:xuefanone@163.com 第5期黄欣然等:脑多肽对PCPA诱导的睡眠障碍小鼠5-羟色胺系统和肠道菌群的影响河北大学学报(自然科学版) 第44卷neurotransmitter levels and oxidative stress markers. Hematoxylin-eosin staining(H&E)and glycogen staining(AB-PAS)were used to observe histopathological changes in the hippocampus and colon of mice. The stool was collected and 16S rRNA gene was sequenced to analyze the changes of intestinal flora abundance in mice. Compared with the model group, the sleep latency of mice in the high-dose and low-dose brain polypeptide groups was significantly shortened, and the sleep time was significantly increased. MDA content decreased significantly, 5-HT, GABA and MT content, the enzyme activity of GSH-Px and SOD increased significantly in high-dose brain polypeptide group(P<0.05). The neuronal cells in CA1 region of hippocampus of mice were arranged neatly, the phenomenon of nuclear shrinkage and deep staining was improved, and the infiltration of inflammatory cells in colon tissue was reduced. 16S rRNA results showed that compared with the model group, the abundance of Bacteroidetes was significantly increased in the high-dose BP group at the phylum level, and the abundance of Lactobacillus and Bifidobacterium at the genus level was significantly increased(P<0.05). Pearson correlation analysis showed that the abundances of Bacteroides was positively correlated with the trends of GSH-Px, and the abundances of Lactobacillus was positively correlated with the trend of 5-HT, MT and SOD(P<0.05). Brain polypeptide can effectively treat PCPA induced insomnia in mice by regulating intestinal flora and positively regulating serum neurotransmitter and brain tissue oxidative stress level, and the therapeutic effect is better with the increase of dose.
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