桂枝茯苓丸对子宫腺肌病小鼠肠道菌群和代谢物的调节作用研究  

Study on Modulation of Gut Microbiota and Metabolites by Guizhi Fuling Pill(桂枝茯苓丸)in Mice with Adenomyosis

作　　者：刘士丹 陈佩佩 付先芸[2] 魏绍斌[1,3] 王坤[2] 陈敏敏 LIU Shidan;CHEN Peipei;FU Xianyun;WEI Shaobin;WANG Kun;CHEN Minmin(TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province,Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,Sichuan,China;Third-Grade Pharmacological Laboratory on Chinese Medicine of National Administration of Traditional Chinese Medicine,College of Medicine and Health Sciences,China Three Gorges University,Yichang 443002,Hubei,China;Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,Sichuan,China)

机构地区：[1]成都中医药大学附属医院代谢性疾病中医药调控四川省重点实验室,四川成都610072 [2]三峡大学国家中医药管理局中药药理三级实验室,三峡大学健康医学院,湖北宜昌443002 [3]成都中医药大学附属医院,四川成都610072

出　　处：《中华中医药学刊》2025年第4期155-161,I0031-I0036,共13页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81973897);成都中医药大学附属医院代谢性疾病中医药调控四川省重点实验室开放基金项目(SZKF202206)。

摘　　要：目的联合肠道菌群组学和代谢组学技术探索桂枝茯苓丸(Guizhi Fuling Pill,GFW)干预子宫腺肌病(adenomyosis,AM)的潜在机制。方法7周龄雌性ICR小鼠随机分为对照组、模型组和GFW组,采用同种异体垂体移植法造模,3组分别灌胃给药8周后处死。评价小鼠子宫组织病理学变化并检测离体子宫收缩频率和波幅。采用16S rRNA基因测序技术对比GFW干预前后肠道菌群Alpha及Beta多样性变化,筛选差异肠道菌群。采用液相色谱-质谱(liquid chromatography-mass spectrometry,LC-MS)技术探索GFW干预后肠道代谢产物的变化。对差异菌群及差异代谢物进行相关性分析,寻找GFW调控AM的可能机制,并进行实验验证。结果HE染色结果证实成功形成AM模型。与模型组相比,给予GFW治疗后体外子宫收缩波幅明显降低(P<0.05),收缩频率呈下降趋势。肠道菌群Alpha多样性显示3组样本间的Observed species指数、Ace指数差异均无统计学意义;Beta多样性结果表明,对照组和模型组样本肠道菌群分布有显著差异,GFW治疗后能一定程度回调AM小鼠的肠道菌群组成。进一步研究表明,AM小鼠肠道中拟杆菌门、拟杆菌属丰度显著下调(P<0.01,P<0.01),厚壁菌门、柔壁菌门相对丰度显著升高(P<0.05,P<0.05)。经GFW干预后,柔壁菌门、支原体属以及肠杆菌属丰度显著降低(P<0.05,P<0.05,P<0.01),而副拟杆菌属相对丰度显著升高(P<0.05)。代谢组学筛选结果证实AM模型小鼠具有22种与氧化应激、免疫及子宫平滑肌收缩等相关的肠道差异代谢物,如前列腺素D_(3)、谷氨酸、色氨酸、羟基二十碳四烯酸、粪胆素等。相关性分析显示,部分肠道菌群与代谢物之间存在显著相关性。实验验证结果证实,AM小鼠子宫组织中存在活性氧(reactive oxygen species,ROS)及肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)、白介素-17(interleukin 17,IL-17)表达水平的上调,经GFW治疗后ROS及TNF-α的表达水平�Objective To explore the potential mechanism of Guizhi Fuling Pill(桂枝茯苓丸,GFW) in the treatment of adenomyosis(AM) by combining gut microbiota and metabolomics technology. Methods Seven-week-old ICR mice were randomized assigned to the control, model and GFW groups. The AM model was developed by the allogeneic pituitary transplantation method. The three groups were sacrificed after 8 weeks of intragastric administration. Histopathological changes of the mice uterus were evaluated, and the frequency and amplitude of uterine contraction in vitro were detected. 16S rRNA gene sequencing was utilized to compare the changes in the Alpha and Beta diversity of the intestinal flora before and after GFW intervention, and to screen the differential gut microbiota. The changes of gut metabolites after GFW treatment were explored using liquid chromatography-mass spectrometry(LC-MS). Correlation analysis was performed on the differential microbiota and metabolites to find out the possible mechanism of GFW regulating AM,followed by experimental validation. Results HE staining confirmed that the AM model was successfully formed. Compared with those of the model group, the amplitude of uterine contraction in vitro was significantly decreased after GFW treatment(P<0.05),and the contraction frequency showed a downward trend. The Alpha diversity of gut microbiota showed that there was no significant difference in Observed species and Ace index among the three groups. The distribution of gut microbiota varied significantly between the control and model groups, according to the Beta diversity results. GFW treatment was able to modulate the gut flora composition of AM mice to varying degrees. Further analysis indicated that the abundance of Bacteroidetes and Bacteroides in the intestine of AM mice was significantly down-regulated(P<0.01,P<0.01),whereas the relative abundance of Firmicutes and Tenericutes was significantly up-regulated(P<0.05,P<0.05). After GFW intervention, the abundance of Tenericutes,Mycoplasma and Enterorha

关 键 词：子宫腺肌病 桂枝茯苓丸 肠道菌群组学 代谢组学 

分 类 号：R289.5[医药卫生—方剂学]