Effects of brain peptides on 5-hydroxytryptamine system and intestinal flora in PCPA induced sleep disorder mice

doi:10.3969/j.issn.1000-1565.2024.05.005

Abstract

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- 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.

Key words: brain polypeptide, 5-hydroxytryptamine, intestinal flora, oxidative stress, somnipathy

CLC Number:

R917

HUANG Xinran, WANG Linna, DAI Jiaqi, SONG Nannan, ZHOU Chengyan. Effects of brain peptides on 5-hydroxytryptamine system and intestinal flora in PCPA induced sleep disorder mice[J]. Journal of Hebei University(Natural Science Edition), 2024, 44(5): 483-494.

References

[1] 郁阿翠,钟云,阮晓东.脑肠肽在睡眠障碍中的作用研究进展[J].新乡医学院学报, 2018, 35(3): 248-251. DOI: 10.7683/xxyxyxb.2018.03.022.
[2] KRAUS S S, RABIN L A. Sleep America: managing the crisis of adult chronic insomnia and associated conditions[J]. J Affect Disord, 2012, 138(3): 192-212. DOI: 10.1016/j.jad.2011.05.014.
[3] ISAIAH S, LOOTS D T, SOLOMONS R, et al. Overview of brain-to-gut axis exposed to chronic CNS bacterial infection(s)and a predictive urinary metabolic profile of a brain infected by Mycobacterium tuberculosis[J]. Front Neurosci, 2020, 14: 296. DOI: 10.3389/fnins.2020.00296.
[4] REYNOLDS A C, PATERSON J L, FERGUSON S A, et al. The shift work and health research agenda: considering changes in gut microbiota as a pathway linking shift work, sleep loss and circadian misalignment, and metabolic disease[J]. Sleep Med Rev, 2017, 34: 3-9. DOI: 10.1016/j.smrv.2016.06.009.
[5] 陈天赐,武少兰,杨国辉,等.无柄灵芝醇提物对小鼠睡眠及肠道菌群的影响[J].生物技术通报, 2022, 38(8): 225-232. DOI: 10.13560/j.cnki.biotech.bull.1985.2021-1470.
[6] LIU Y T, CHENG F Y, TAKEDA S, et al. Effects of porcine brain hydrolysate on impairment of cognitive learning ability in amyloid β(1-40)-infused rats[J]. Anim Sci J, 2019, 90(2): 271-279. DOI: 10.1111/asj.13054.
[7] 龙盼,胡晓霞,胡琦兰,等.天麻素对氯苯丙氨酸(PCPA)致失眠小鼠的镇静催眠作用研究[J].中药药理与临床, 2021, 37(5): 33-38. DOI: 10.13412/j.cnki.zyyl.2021.05.007.
[8] 谢懂君,吴旭玲,董楝,等.甘露特钠对慢性癫痫大鼠认知功能障碍的影响[J].中国神经精神疾病杂志, 2023, 49(6): 351-356. DOI: 10.3969/j.issn.1002-0152.2023.06.006.
[9] 姚玉静,张书敏,任艳艳,等.酸枣仁提取物、龙眼肉提取物、γ-氨基丁酸和酪蛋白水解物复配制剂改善睡眠功能[J].食品工业科技, 2023, 44(7): 406-410. DOI: 10.13386/j.issn1002-0306.2022100103.
[10] 刘玉凤,黄枚,南丽红,等.古方半夏秫米汤对正常小鼠最大给药量及其对失眠模型大鼠作用的实验研究[J].中药药理与临床, 2023, 39(8): 12-18. DOI: 10.13412/j.cnki.zyyl.20230524.007.
[11] 张振宇,米传靓,李思迪,等.基于16S rRNA测序分析脂多糖结合蛋白基因敲除对小鼠肠道菌群的影响[J].中国医药生物技术, 2023, 18(2): 110-115. DOI: 10.3969/j.issn.1673-713X.2023.02.003.
[12] 侯金良,张媛媛,张浩,等.沉香片剂小鼠口鼻吸入给药抗焦虑和催眠功效实验研究[J].山东中医药大学学报, 2021, 45(1): 113-119. DOI: 10.16294/j.cnki.1007-659x.2021.01.018.
[13] 高荣静,黄红,姜宁,等.大豆异黄酮对脂多糖诱导小鼠抑郁样行为的改善作用研究[J].大豆科学, 2021, 40(4): 539-545. DOI: 10.11861/j.issn.1000-9841.2021.04.0539.
[14] 王瑞英,苏丹,李惠珍,等.生、酒五味子对失眠小鼠神经-内分泌-免疫网络的影响及机制[J].中国药房, 2023, 34(5): 525-530. DOI: 10.6039/j.issn.1001-0408.2023.05.03.
[15] 范恩宇,魏振华,范秋领,等.黑柄炭角菌(乌灵菌)粉改善睡眠作用以及对小鼠大脑神经递质及其受体的影响[J].菌物学报, 2022, 41(9): 1498-1505. DOI: 10.13346/j.mycosystema.210491.
[16] 徐飞飞,田雅娟,李钦青,等.酸枣仁-茯苓-党参水提物对小鼠睡眠的改善作用及机制研究[J].食品工业科技, 2021, 42(11): 300-308. DOI: 10.13386/j.issn1002-0306.2020070005.
[17] 李莉,刘茹,何晶,等.温胆汤方剂对睡眠剥夺小鼠氧化应激及炎症反应的影响[J].中国免疫学杂志, 2022, 38(3): 308-312. DOI: 10.3969/j.issn.1000-484X.2022.03.009.
[18] 李欣益,刘春妍,黄建,等.益生菌V9对高脂饮食诱导的NAFLD大鼠肝功能、氧化应激及脂代谢的影响及作用机制[J].中国免疫学杂志, 2019, 35(23): 2822-2826. DOI: 10.3969/j.issn.1000-484X.2019.23.002.
[19] 郭鹏.左旋丁基苯酞对血管性痴呆大鼠神经功能、线粒体氧化应激及海马组织胶质细胞源性神经营养因子表达的影响[J].中国老年学杂志, 2019, 39(6): 1444-1447. DOI: 10.3969/j.issn.1005-9202.2019.06.052.
[20] 刁华琼,魏丹,丁海月,等.黄连阿胶汤对睡眠剥夺大鼠5-羟色胺系统和肠道菌群的影响[J].中国实验方剂学杂志, 2023, 29(21): 49-58. DOI: 10.13422/j.cnki.syfjx.20231043.
[21] 万叶敏.基于肠-脑轴PI3K/AKT/mTOR自噬通路探讨加味通便汤改善STC的临床疗效及作用机制[D].南京:南京中医药大学, 2022. DOI:10.27253/d.cnki.gnjzu.2022.000029.
[22] BEAR T L K, DALZIEL J E, COAD J, et al. The role of the gut microbiota in dietary interventions for depression and anxiety[J]. Adv Nutr, 2020, 11(4): 890-907. DOI: 10.1093/advances/nmaa016.
[23] 陈抒鹏,唐娜娜,王思梦,等.肝郁脾虚方对抑郁失眠模型大鼠结肠组织GABA能信号系统及肠道菌群的影响[J].中医杂志, 2023, 64(12): 1273-1281. DOI: 10.13288/j.11-2166/r.2023.12.014.
[24] 屈婉,刘硕,朱华伟,等.酸枣仁水提物对焦虑大鼠症状的改善及其对肠道菌群的影响[J].现代食品科技, 2019, 35(11): 30-36. DOI: 10.13982/j.mfst.1673-9078.2019.11.005.
[25] 杨烨,徐兴军,李浩雨,等.罗汉果山柰苷对慢性睡眠剥夺小鼠抗氧化能力的影响[J].动物营养学报, 2023, 35(12): 8083-8096. DOI: 10.12418/CJAN2023.732.
[26] 陈雪梅,胡博,唐晓姝,等.紫苏籽油对小鼠肠道微生物及抗氧化能力的影响[J].食品与发酵工业,2024,50(16):271-277.DOI: 10.13995/j.cnki.11-1802/ts.037422.
[27] 任璐婷,夏满盈,罗瑞琳,等.苹果发酵物对溃疡性结肠炎小鼠肠道菌群的调节作用[J].食品与发酵工业,2024,50(15):33-40.DOI: 10.13995/j.cnki.11-1802/ts.039431. (