摘要
目的:该研究旨在整合非靶向代谢组学与靶向短链脂肪酸(SCFAs)代谢组学探究白鲜皮对大鼠肠道损伤的影响及其机制。方法:将大鼠随机分为空白组、白鲜皮高剂量组(8.1 g·kg^(-1))、中剂量组(2.7 g·kg^(-1))、低剂量组(0.9 g·kg^(-1)),除空白组外,白鲜皮各给药组灌胃不同剂量白鲜皮提取物,连续8周;苏木素-伊红(HE)染色观察回肠组织病理变化,酶联免疫吸附测定法(ELISA)检测大鼠回肠组织中细胞因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)水平,采用实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠回肠组织中紧密连接蛋白闭锁小带蛋白-1(ZO-1)、闭合蛋白(Occludin)和密封蛋白-1(Claudin-1)mRNA表达水平,初步探究白鲜皮对肠道损伤的作用。选取毒性表型最为显著的剂量采用非靶向代谢组学联合靶向短链脂肪酸代谢组学技术进一步揭示白鲜皮对大鼠回肠组织代谢轮廓的影响。结果:与空白组比较,白鲜皮各剂量组均可诱发不同程度的回肠病理损伤,升高回肠组织中TNF-α、IL-6、IL-1β水平(P<0.01),降低回肠组织中ZO-1(P<0.05,P<0.01)、Occludin(P<0.01)和Claudin-1(P<0.05)mRNA表达水平,其中以高剂量组毒性表型最为显著;整合非靶向代谢组学与靶向短链脂肪酸代谢组学技术进一步探究白鲜皮高剂量组对回肠组织的损伤机制:非靶向代谢结果显示,空白组与白鲜皮高剂量组共同鉴定出21个差异代谢物,其中与空白组比较,白鲜皮干预后可显著升高回肠内容物中14个代谢物水平(P<0.05,P<0.01),显著降低回肠内容物中7个代谢物水平(P<0.05,P<0.01),以上代谢物共同作用于甘油磷脂、初级胆汁酸生物合成等10条相关代谢途径;靶向SCFAs代谢组学定量数据结果显示,白鲜皮干预后可扰乱大鼠回肠内容物样本中丙酸、丁酸、乙酸、己酸、异丁酸、异戊酸、戊酸及异己酸水平,与空白组比较,白鲜皮干预后大鼠回肠内容物中异丁酸、异戊酸、戊酸水平显著升高(P<0.01),丙酸、丁酸、乙酸水平明显降低(P<0.05,P<0.01)。结论:白鲜皮可诱发肠道损伤,具体机制为通过调控甘油磷脂代谢、初级胆汁酸生物合成代谢及SCFAs代谢途径进而诱发肠道损伤。
Objective:This study aims to investigate the effect of Dictamni Cortex on intestinal barrier damage in rats and its mechanism by untargeted metabolomics and targeted metabolomics for short-chain fatty acids(SCFAs).Methods:Rats were randomly divided into a control group,a high-dose group of Dictamni Cortex(8.1 g·kg^(-1)),a medium-dose group(2.7 g·kg^(-1)),and a low-dose group(0.9 g·kg^(-1)).Except for the control group,the other groups were administered different doses of Dictamni Cortex by gavage for eight consecutive weeks.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in the ileal tissue.Enzyme-linked immunosorbent assay(ELISA)was employed to detect the level of cytokines,including tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β),in the ileal tissue of rats.Quantitative real-time fluorescence polymerase chain reaction(Real-time PCR)technology was used to detect the expression level of tight junction proteins,including zonula occludens-1(ZO-1),Occludin,and Claudin-1 mRNAs,in the ileal tissue of rats to preliminarily explore the effects of Dictamni Cortex on intestinal damage.The dose with the most significant toxic phenotype was selected to further reveal the effects of Dictamni Cortex on the metabolic profile of ileal tissue in rats by non-targeted metabolomics combined with targeted metabolomics for SCFAs.Results:Compared with the control group,all doses of Dictamni Cortex induced varying degrees of pathological damage in the ileum,increased TNF-α(P<0.01),IL-6(P<0.01),and IL-1β(P<0.01)levels in the ileal tissue,and decreased the expression level of ZO-1(P<0.05,P<0.01),Occludin(P<0.01),and Claudin-1(P<0.05)in the ileal tissue,with the high-dose group showing the most significant toxic phenotypes.The damage mechanisms of the high-dose group of Dictamni Cortex on the ileal tissue were further explored by integrating non-targeted metabolomics and targeted metabolomics for SCFAs.The non-targeted metabolomics results showed that 21 differential metabolites were identified in the control group and the high-dose group.Compared with that in the control group,after Dictamni Cortex intervention,the level of 14 metabolites was significantly increased(P<0.05,P<0.01),and the level of seven metabolites was significantly decreased(P<0.05,P<0.01)in the ileal contents.These metabolites collectively acted on 10 related metabolic pathways,including glycerophospholipids and primary bile acid biosynthesis.The quantitative data of targeted metabolomics for SCFAs showed that Dictamni Cortex intervention disrupted the level of propionic acid,butyric acid,acetic acid,caproic acid,isobutyric acid,isovaleric acid,valeric acid,and isocaproic acid in the ileal contents of rats.Compared with those in the control group,the level of isobutyric acid,isovaleric acid,and valeric acid were significantly increased,while the level of propionic acid,butyric acid,and acetic acid were significantly decreased in the ileal contents of rats after Dictamni Cortex intervention(P<0.05,P<0.01).Conclusion:Dictamni Cortex can induce intestinal damage by regulating glycerophospholipid metabolism,primary bile acid biosynthesis,and metabolic pathways for SCFAs.
作者
徐晓敏
于栋华
王宇
陈平平
沃佳美雪
贾素霞
胡文凯
卢芳
刘树民
XU Xiaomin;YU Donghua;WANG Yu;CHEN Pingping;WO Jiameixue;JIA Suxia;HU Wenkai;LU Fang;LIU Shumin(College of Traditional Chinese Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《中国实验方剂学杂志》
北大核心
2025年第20期40-47,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家重点研发计划“中医药现代化”重点专项(2022YFC3502100)。
关键词
白鲜皮
肠屏障功能
非靶向代谢组学
靶向短链脂肪酸代谢组学
Dictamni Cortex
intestinal barrier function
untargeted metabolomics
targeted metabolomics for short-chain fatty acid
