摘要
目的观察补阳还五汤对大鼠脑缺血再灌注损伤模型肠道菌群的影响。方法使用OMEGA Soil DNA Kit提取样品核酸。细菌项目选取约468bp的16S rRNA V3V4区进行测序。PCR扩增采用16S rRNA V3-V4区特异性引物338F(5'barcode+ACTCCTACGGGAGGCAGCA-3')和806R(5'-GGACTACHVGGGTWTCTAAT-3')。利用Tru Seq Nano DNALT Library Prep Kit建库并质检,对合格文库进行2×250bp双端测序。结果1)模型组大鼠脑组织有明显的梗死,给药后梗死面积显著减少。2)不同样本的菌群结构发生明显变化,对照组菌群物种属水平中乳酸菌丰度最高,模型组菌群物种属水平中乳酸菌丰度明显降低,Akkermansiaj菌丰度相对偏高,给药组菌群物种属水平中仍以Baoteroides菌、Allobaculum菌的丰度为主,但是乳酸菌和杆菌丰度明显升高。3)不同样本(组)大鼠ASV/OTU序列部分重叠,但每组都有一些独特的ASV/OTU序列,如对照组有4648个,模型组有2126个,实验组有2308个。在给药组中,梭状芽孢杆菌是导致与其他两组差异显著的最大贡献因素,模型组中肠球菌科是导致与其他两组差异显著的最大贡献因素,对照组中厚壁菌门是导致与其他两组差异显著的最大贡献因素。4)对照组少部分面积与模型组重合,仅极少部分功能单元3组重合,且模型组、治疗组离对照组远。结论补阳还五汤可促进肠道乳酸因子和双歧杆菌的再生,并且使其益生菌功能更集中。
Objective:To investigate the effect of Buyang Huanwu Decoction on the intestinal flora in a rat model of cerebral ischemia-reperfusion injury.Methods:Genomic DNA was extracted from fecal samples using the OMEGA Soil DNA Kit.The V3~V4 hypervariable regions(about 468 bp)of bacterial 16S rRNA genes were amplified by PCR using specific primers 338F(5'-barcode-ACTCCTACCGGAGGCAGCA-3')and 806R(5'-GGACTACHVGGGTWTCTAAT-3').Libraries were constructed using the TruSeq Nano DNA LT Library Prep Kit,followed by quality control and paired-end sequencing(2×250 bp)for qualified libraries.Results:1)The model group showed obvious cerebral infarction,which was significantly reduced after administration of the decoction.2)Significant changes were observed in the structure of the gut microbiota among groups.At the genus level,Lactobacillus was most abundant in the control group,while its abundance was significantly decreased in the model group,accompanied by an increase in Akkermansia.In the treatment group,Bacteroides and Allobaculum remained dominant,but the abundances of Lactobacillus and Bifidobacterium were notably increased.3)Although some ASVs/OTUs overlapped among dfferent groups,each group also had unique sequences.For example,the control group had 4648 unique ASVs/OTUs,the model group had 2126,and the treatment group had 2308.In the treatment group,Clostridium was the main contributor to the significant differences compared with the other two groups.In the model group,Enterococcaceae contributed most to the differences,while in the control group,Firmicutes was the key differential factor.4)The control group partially overlapped with the model group in a few functional units,but only a small number of functional units overlapped among all three groups.Moreover,both the model and treatment groups were far from the control group in clustering analysis.Conclusion:Buyang Huanwu Decoction promotes the regeneration of intestinal Lactobacillus and Bifidobacterium,enhancing their probiotic functions and making them more concentrated.
作者
刘志勇
蒋建强
王更磊
于静华
Liu Zhiyong;Jiang Jianqiang;Wang Genglei;Yu Jinghua(Kaifeng Second Traditional Chinese Medicine Hospital,Henan,Kaifeng 475000,China)
出处
《中国中医急症》
2025年第7期1152-1157,1163,共7页
Journal of Emergency in Traditional Chinese Medicine
基金
开封市科技发展计划项目(2303121)。
关键词
缺血性中风
气虚血瘀证
补阳还五汤
肠道菌群
大鼠
Ischemic stroke
Syndrome of blood stasis due to qi deficiency
Buyang Huanwu Decoction
Intestinal flora
Rats
