黄芪囊泡样纳米颗粒通过调节肠道菌群降低db/db糖尿病小鼠血糖的作用机制分析  

Mechanism of Astragali Radix Vesicle-like Nanoparticles for Reducing Blood Glucose in db/db Diabetic Mice by Regulating Gut Microbiota

作　　者：高文静 侯敏 陈潇潇 王攀 任钧国[1] 刘建勋[1] GAO Wen-jing;HOU Min;CHEN Xiao-xiao;WANG Pan;REN Jun-guo;LIU Jian-xun(Beijing Key Laboratory of Pharmacology of Traditional Chinese Medicine,Institute of Basic Medical Sciences,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,中药药理北京市重点实验室,北京100091 [2]北京中医药大学,北京100029

出　　处：《中国实验方剂学杂志》2021年第14期111-118,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家“重大新药创制”科技重大专项(2017ZX09301018);中国中医科学院基本科研业务费自主选题(ZZ11-027,zz110324)。

摘　　要：目的:研究黄芪水煎液中囊泡样纳米颗粒(VLNs)的提取方法及特性,探讨黄芪VLNs通过调节db/db小鼠肠道菌群来降低血糖的作用机制。方法:采用超速离心法和尺寸排阻色谱法分别富集黄芪水煎液中的VLNs,通过透射电子显微镜、纳米颗粒跟踪分析仪分析黄芪水煎液中VLNs的形态、粒径、浓度。选用db/db糖尿病小鼠,按血糖值随机分为模型组,黄芪VLNs高、中、低剂量(21.1,10.6,5.3 g·kg^(-1))组和二甲双胍组(0.25 g·kg^(-1)),每组7只,另设7只C57BL/6小鼠作为正常组,灌胃给药3周(每日1次),每周观察小鼠空腹血糖变化;苏木素-伊红(HE)染色观察糖尿病小鼠肝脏和胰腺病理形态情况;采集小鼠粪便进行肠道微生物16S rRNA多样性检测。结果:2种方法所得纳米颗粒粒径均在200 nm左右,超速离心法提取的黄芪VLNs具有典型的茶托状,其浓度为3.0×10^(11)个/mL;尺寸排阻色谱法所得黄芪VLNs形态相对较差,其浓度为2.2×10^(11)个/mL。给药3周后,与模型组比较,黄芪VLNs高、中、低剂量组均能显著降低糖尿病小鼠的空腹血糖(P<0.05,P<0.01);可改善肠道菌群失调,显著降低厚壁菌门/拟杆菌门比例,提高有益菌相对丰度,降低有害菌相对丰度。结论:黄芪VLNs可能通过调节肠道菌群中厚壁菌门/拟杆菌门的比例来降低db/db糖尿病小鼠的血糖。Objective:To study the extraction method and characteristics of vesicle-like nanoparticles(VLNs)in Astragali Radix decoction,and to explore the mechanism of the VLNs in reducing blood glucose by regulating the gut microbiota of db/db diabetic mice.Method:Ultracentrifugation and size exclusion chromatography were used to enrich VLNs from Astragali Radix decoction,and the morphology,particle size and concentration of the VLNs were analyzed by transmission electron microscope and nanoparticle tracking analyzer.The db/db diabetic mice were randomly divided into model group,Astragali Radix VLNs high-,medium-and low-dose(21.1,10.6,5.3 g·kg^(-1))groups and metformin group(0.25 g·kg^(-1))according to their blood glucose levels.There were 7 mice in each group,and another 7 C57BL/6 mice were set as the normal group.The mice were given intragastrically for 3 weeks(once a day),and the changes of fasting blood glucose were observed every week.Hematoxylin-eosin(HE)staining was used to observe the pathological morphology of liver and pancreas of diabetic mice.The feces of mice were collected for 16 S rRNA diversity detection of intestinal microbes.Result:The size of the nanoparticles obtained by the two methods was about 200 nm.Astragali Radix VLNs extracted by ultracentrifugation had a typical saucer-like shape with the concentration of 3.0×10^(11)particles·mL-1.The morphology of Astragali Radix VLNs obtained by size exclusion chromatography was relatively poor with the concentration of 2.2×10^(11)particles·mL-1.After 3 weeks of administration,compared with the model group,Astragali Radix VLNs high-,medium-and low-dose groups could significantly reduce the fasting blood glucose of diabetic mice(P<0.05,P<0.01).The VLNs could improve the gut microbiota dysbiosis,significantly decrease the ratio of Firmicutes/Bacteroidota,and increase the relative abundance of beneficial bacteria and reduce the relative abundance of harmful bacteria.Conclusion:Astragali Radix VLNs may reduce the blood glucose of db/db diabetic mice by adjus

关 键 词：黄芪 囊泡样纳米颗粒 超速离心法 尺寸排阻色谱法 空腹血糖 肠道菌群 粒径分布 

分 类 号：R587[医药卫生—内分泌] R94[医药卫生—内科学]