基于Th17/Treg免疫平衡研究黄芩醇提物治疗溃疡性结肠炎的作用机制
Exploration of mechanism of ethanol extract of Scutellaria baicalensis Georgi in treating ulcerative colitis based on Th17/Treg immune balance
郭春雨 1董红敬 2郭兰萍 3李佳 4潘少斌 4李庆军 4王晓2
作者信息
- 1. 山东中医药大学药学院,济南 250300;齐鲁工业大学(山东省科学院)山东省分析测试中心山东省大型精密分析仪器应用技术重点实验室,济南 250014
- 2. 齐鲁工业大学(山东省科学院)山东省分析测试中心山东省大型精密分析仪器应用技术重点实验室,济南 250014
- 3. 中国中医科学院中药资源中心道地药材国家重点实验室培育基地,北京 100700
- 4. 山东中医药大学药学院,济南 250300
- 折叠
摘要
目的:探究黄芩醇提物调节免疫平衡治疗溃疡性结肠炎(ulcerative colitis,UC)的作用及机制.方法:将24 只C57BL/6 小鼠随机分为正常组、模型组、美沙拉嗪组(500 mg·kg-1)、黄芩醇提物(400 mg·kg-1)组,每组6 只.采用2.5%葡聚糖硫酸钠(dextran sulfate sodium,DSS)的饮用水建立UC小鼠模型.每天观察各组小鼠体重及一般情况,d6 计算疾病活动指数(DAI);测量结肠长度;采用流式细胞术检测肠系膜淋巴结和脾脏中辅助性T细胞17(T helper 17,Th17)和调节性T细胞(regulatory T cells,Treg)比例;苏木精-伊红染色观察结肠组织损伤程度;酶联免疫吸附法检测结肠白细胞介素-6(interleukin,IL-6),IL-10 和IL-1β的含量;蛋白免疫印迹法检测结肠Toll样受体4(Toll-like receptor 4,TLR4)、核因子κB p65(nuclear factor-kappa B,NF-κB p65)、p-NF-κB p65 及核因子κB抑制蛋白-α(I kappaB-alpha,IκB-α)的表达水平;气相色谱-质谱联用检测短链脂肪酸的含量;高内涵成像技术观察RAW264.7 细胞核转位情况.结果:黄芩醇提物可改善DSS诱导的小鼠结肠炎症状,恢复肠系膜淋巴结和脾脏中Th17/Treg免疫平衡,降低结肠组织中IL-1β和IL-6 水平、提高IL-10 表达水平,降低TLR4,NF-κB p65 和p-NF-κB p65 表达水平,提高IκB-α表达水平,提高肠道短链脂肪酸含量,抑制脂多糖诱导引起的核转位.结论:黄芩醇提物可以改善UC小鼠的症状,其机制可能是通过改善肠道稳态,抑制TLR4/NF-κB通路以恢复Th17/Treg免疫平衡.
Abstract
Objective:To explore the effect and mechanism of alcohol extract of Scutellaria baicalensis Georgi on immune balance in treating ulcerative colitis(UC).Methods:24 C57BL/6 mice were randomly divided into control group,model group,mesalazine group,and Scutellaria baicalensis Georgi alcohol extract group(400 mg·kg-1,n =6 for each group).A mouse model of UC was established by drinking water containing 2.5%dextran sulfate sodium(DSS).The body weight and general condition of the mice in each group were observed every day,and the disease activity index(DAI)was calculated on the 6th day;the length of the colon was measured;the proportion of T helper 17(Th17)/regulatory T cells(Treg cells)in the mesenteric lymph nodes and spleen was detected by flow cytometry;hematoxylin-eosin staining was used to observe the degree of colon tissue damage;the levels of interleukin-6(IL-6),IL-10 and IL-1β in colon were detected by enzyme-linked immunosorbent assay;the expressions of Toll-like receptor 4(TLR4),nuclear factor-kappa B(NF-κB p65),p-NF-κB p65,and I kappaB-alpha(IκB-α)in colon were detected by Western blot;the main short chain fatty acids were detected by gas chromatography-mass spectrometry;high-content imaging was used to observe the nuclear translocation of RAW264.7 cells.Results:The alcohol extract of Scutellaria baicalensis Georgi improved the symptoms of colitis induced by DSS in mice,restored the Th17/Treg immune balance in mesenteric lymph nodes and spleen,reduced the levels of IL-1β and IL-6 in colon tissue,and increased the expression level of IL-10.It decreased the expressions of TLR4,NF-κB p65 and p-NF-κB p65,increased the expression of IκB-α,increased the content of intestinal short-chain fatty acids,and inhibited the nuclear translocation induced by LPS.Conclusion:The alcohol extract of Scutellaria baicalensis Georgi can improve the symptoms of UC mice,and the mechanism may be to restore Th17/Treg immune balance by improving intestinal homeostasis and inhibiting TLR4/NF-κB pathway.
关键词
黄芩/醇提物/溃疡性结肠炎/Th17/Treg免疫平衡/TLR4/NF-κB信号通路/短链脂肪酸Key words
Scutellaria baicalensis Georgi/ethanol extract/ulcerative colitis/Th17/Treg immune balance/TLR4/NF-κB signal pathway/short-chain fatty acids引用本文复制引用
基金项目
山东省重点研发计划资助项目(2021CXGC010508)
中央本级重大增减支项目(2060302)
齐鲁工业大学(山东省科学院)重大创新专项资助项目(2022JBZ02-04)
山东省医药卫生科技发展计划资助项目(2019WS554)
出版年
2024
NETL
NSTL
万方数据