基于肠道菌群及其代谢物探讨黄芩苷-甘草苷对小鼠脑缺血再灌注损伤的保护作用及机制  

Exploring the protective effect and mechanism of Baicalin-Liquiritin on cerebral ischaemia-reperfusion injury in mice based on intestinal flora and its metabolites

作　　者：翁明倩 吴冰 刘彩玉 阮君山 王大海 Weng Mingqian;Wu Bing;Liu Caiyu;Ruan Junshan;Wang Dahai(School of Pharmacy,Fujian University of Traditional Chinese Medicine,Fujian,Fuzhou 350122,China;Cadre Speciality ClinicⅡ,Fuzhou University Affiliated Provincial Hospital,Fujian,Fuzhou 350001,China;Department of Pharmacy,Fuzhou University Affiliated Provincial Hospital,Fujian,Fuzhou 350001,China)

机构地区：[1]福建中医药大学药学院,福建福州350122 [2]福州大学附属省立医院干部特诊二科,福建福州350001 [3]福州大学附属省立医院药学部,福建福州350001

出　　处：《创伤与急诊电子杂志》2024年第4期244-260,共17页Journal of Trauma and Emergency(Electronic Version)

基　　金：福建省自然科学基金资助项目(2020J011100);福建省科技重大专项专题项目(2022NZ029017)。

摘　　要：目的通过肠道菌群及其代谢物,探究黄芩苷-甘草苷(Baicalin-Liquiritin,BA-LI)对脑梗死的保护作用和机制,为有效干预脑缺血再灌注损伤提供新的思路。方法将60只小鼠按照随机数字表法分成Sham组(实行假手术)、Model组(仅造模)、NI组(尼莫地平给药,10mg/kg)、L组(BA-LI低剂量给药,BA:127.5mg/kg,LI:6.75mg/kg)、H组(BA-LI高剂量给药,BA:255mg/kg,LI:13.5mg/kg),每组12只,利用线栓法构建大脑中动脉闭塞(middle cerebral artery oocclusion,MCAO)模型。通过2,3,5-氯化三苯基四氮唑(用TTC表示)染色分析、尼氏染色、酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)分析小鼠脑组织损伤情况及炎症因子水平;取粪便进行16S rRNA测序检测肠道菌群变化;短链脂肪酸(short-chain fatty acids,SCFAs)定量分析肠道菌群代谢物中SCFAs的表达。结果H组脑梗死体积比(16.04±2.13)%小于Model组(24.67±1.79)%,差异具有统计学意义(P<0.05);与Model组相比,H组抗炎因子转化生长因子-β、白介素-10表达显著升高,具有统计学意义(P均<0.05)。16S rRNA检测结果显示,BA-LI主要通过调节乳酸杆菌、拟线杆菌等有益菌群丰度,恢复肠道菌群生物多样性。与Model组相比,H组小鼠粪便中乙酸、丁酸、异丁酸的含量增加,差异具有统计学意义(P均<0.05)。结论高剂量的BA-LI对脑缺血再灌注损伤小鼠具有神经保护作用;能调节体内有益菌群,起到减轻脑缺血再灌注损伤的作用。Objective To explore the protective effects and underlying mechanisms of Baicalin-Liquiritin(BA-LI)against cerebral infarction through gut microbiota and its metabolites,and to offer novel insights for the effective intervention of cerebral ischemia-reperfusion injury.Method Sixty mice were randomly allocated into five groups according to the random number table method,namely the sham group undergoing sham operation,the model group subjected only to modeling,the NI group(receiving nimodipine at a dosage of 10 mg/kg),the L group(administered with low-dose BA-LI,wherein BA was 127.5 mg/kg and LI was 6.75 mg/kg),and the H group(administered with high-dose BA-LI,with BA at 255 mg/kg and LI at 13.5 mg/kg),with 12 mice in each cohort.The middle cerebral artery occlusion(MCAO)model was established by the suture-occlusion technique.Subsequently,2,3,5-Triphenyltetrazolium chloride(abbreviated as TTC)staining analysis,Nissl staining,as well as enzyme-linked immunosorbent assay(ELISA)were employed to meticulously evaluate the degree of brain tissue damage and the levels of inflammatory factors in mice.Moreover,fecal samples were collected for 16S rRNA sequencing to detect alterations in gut microbiota.Additionally,quantitative analysis of short-chain fatty acids(SCFAs)was carried out to ascertain the expression profiles of SCFAs within gut microbiota metabolites.Result The cerebral infarction volume ratio in the H group was significantly lower than that in the model group[(16.04±2.13)%v.s.(24.67±1.79)%,P<0.05].In comparison with the model group,the expressions of anti-inflammatory factors,such as transforming growth factor-βand interleukin-10 in the H group,were significantly elevated(all P values<0.05).The 16S rRNA sequencing results indicated that BA-LI predominantly modulated the abundances of beneficial microbial populations,including lactobacillus and bacteroides,thereby restoring the biodiversity of gut microbiota.Moreover,compared with the model group,the contents of acetic acid,butyric acid,and isobutyric acid in

关 键 词：黄芩苷 甘草苷 脑缺血再灌注损伤 肠道菌群 短链脂肪酸 

分 类 号：R28[医药卫生—中药学]