摘要
目的探讨首乌丸延缓脑衰老的可能作用机制。方法将SPF级SD雄性大鼠50只任意分为正常组、模型组、维生素E组、首乌丸组及首乌丸2倍剂量组。除正常组外,其余4组均用D-半乳糖(120 mg/kg)复制衰老模型,同时首乌丸组和首乌丸2倍剂量组分别予首乌丸1.08、2.16 g/kg灌胃,维生素E组予维生素E 0.018 g/kg灌胃。6周造模结束后,各组任意选取6只小鼠进行16S rRNA测序分析肠道菌群多样性和物种丰度的变化,检测海马β-半乳糖苷酶活力,苏木素-伊红(HE)染色观察空肠组织病理形态学变化,用酶联免疫吸附(ELISA)法检测各组大鼠血清及海马的氧化三甲胺(TMAO)含量。Spearman相关性分析基于属水平的差异菌群与血清TMAO的相关性。结果与正常组相比,模型组海马β-半乳糖苷酶活性上升(P<0.01);与模型组相比,首乌丸组和首乌丸2倍剂量组海马β-半乳糖苷酶活性下降(P<0.01)。与模型组相比,首乌丸组和首乌丸2倍剂量组回肠纹状缘比较整齐,黏膜上皮细胞脱落减少。与正常组相比,模型组Chao1指数、Faith指数、Observed_otus指数下降(P<0.05,P<0.01);与模型组相比,首乌丸组Chao1指数、Faith指数、Observed_otus指数、Shannon指数上升(P<0.05,P<0.01),首乌丸2倍剂量组Chao1指数、Shannon指数上升(P<0.05)。肠道菌群Beta多样性分析,各组样本能够区分。门水平肠道菌群,与正常组相比,模型组拟杆菌门、放线菌门相对丰度下降(P<0.01),变形菌门、蓝藻菌门相对丰度上升(P<0.01);与模型组相比,首乌丸组和首乌丸2倍剂量组放线菌门丰度升高(P<0.01),蓝藻菌门丰度降低(P<0.01),厚壁菌门与拟杆菌门相对丰度的比率均降低。属水平,与正常组相比,模型组普雷沃菌属、乳酸杆菌属、颤螺菌属、拟杆菌属、副雷沃菌属相对丰度下降(P<0.01);与模型组相比,经首乌丸干预后,普雷沃菌属、乳酸杆菌属、颤螺菌属、拟杆菌属、副雷沃菌属、布劳特菌属、狄氏副拟杆菌属相对丰度均上升(P<0.01)。与正常组相比,模型组血清及海马TMAO含量升高(P<0.01);与模型组相比,首乌丸组和首乌丸2倍剂量组血清及海马TMAO含量下降(P<0.01)。Spearman相关性分析提示,普雷沃菌属、乳酸杆菌属、副雷沃菌属、布劳特菌属相对丰度与血清TMAO含量呈负相关。结论首乌丸能够降低D-半乳糖衰老大鼠海马β-半乳糖苷酶活性,改善空肠病理形态,其作用机制可能是通过调节肠道菌群,减少TMAO的生成,进而延缓脑衰老。
Objective To explore the possible mechanism of Shouwu Pill in delaying brain aging.Methods Fifty male SPF SD rats were randomly divided into the normal group,model group,vitamin E group,Shouwu Pill group,and double dose Shouwu Pill group.Except the normal group,the other four groups were used to generate aging models with D-galactose(120 mg/kg).Simultaneously,Shouwu Pill was used to intervene the Shouwu Pill group and double dose Shouwu Pill group(1.08,2.16 g/kg),while the vitamin E group(0.018 g/kg)was given vitamin E by gavage.After 6 weeks of modeling,six mice per group were randomly selected for 16S rRNA sequencing to analyze the changes in intestinal flora diversity and species richness,and to detect the hippocampusβ-galactosidase activity.Hematoxylin and eosin(HE)staining were used to observe the pathological changes of jejunum tissue,and the content of trimethylamine oxide(TMAO)in serum and hippocampus of rats in each group was detected by enzyme-linked immunosorbent assay(ELISA).Results Compared to the normal group,theβ-galactosidase activity increased in the hippocampus of the model group(P<0.01).Compared to the model group,theβ-galactosidase activity decreased in the hippocampus of the Shouwu Pill group and double dose Shouwu Pill group(P<0.01).Compared to the model group,the striated border of the ileum in the Shouwu Pill group and double dose Shouwu Pills group was neat,and the shedding of mucosal epithelial cells was reduced.Compared to the normal group,the model group showed a decreased Chao1 index,Faith index,and Observed_otus(P<0.05,P<0.01).Compared to the model group,the Chao1 index,Faith index,Observed_otus,and Shannon index(P<0.05,P<0.01)increased in the Shouwu Pill group,and the Chao1 index and Shannon index in the double dose group increased(P<0.05).Beta diversity analysis of intestinal flora demonstrated that each group of samples could be significantly distinguished.At the phyla level,compared to the normal group,the abundance of Bacteroides and Actinomycetes in the model group decreased(P<0.01),while the abundance of Proteus and Cyanobacteria in the model group increased(P<0.01).Compared to the model group,the ratio of relative abundance of Firmicutes and Bacteroides in the Shouwu Pill group and double dose Shouwu Pill group decreased,the abundance of Actinomycetes increased(P<0.01),and the abundance of Cyanobacteria decreased(P<0.01).At the genus level,compared to the normal group,the relative abundance of Prevotella,Lactobacillus,Vibrio,Bacteroides,and Para Prevotella in the model group decreased(P<0.01).Compared to the model group,after the intervention of Shouwu Pill,the relative abundance of Prevotella,Lactobacillus,Vibrio,Bacteroides,Para Prevotella,Brautzia,and Para Prevotella increased(P<0.01).Compared to the normal group,the content of TMAO in the serum and hippocampus of the model group was significantly increased(P<0.01).Compared to the model group,the content of TMAO in the serum and hippocampus in the Shouwu Pill group and double dose Shouwu Pill group were significantly decreased(P<0.01).Spearman correlation analysis showed that Prevotella,Lactobacillus,Parapre votella,and Brautzia were negatively correlated with serum TMAO.Conclusion Shouwu Pill can reduce theβ-galactosidase activity in the hippocampus of aging rats treated with D-galactose.Moreover,theβ-galactosidase activity can improve the pathological morphology of the jejunum,which may delay brain aging by regulating intestinal flora and reducing the production of TMAO.
作者
张超
加秀凤
黄万凌
周勇
汤琪
陈刚
ZHANG Chao;JIA Xiufeng;HUANG Wanling;ZHOU Yong;TANG Qi;CHEN Gang(College of Basic Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;Department of Chinese Medicine,Hubei College of Chinese Medicine,Jingzhou 434020,China;Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430017,China;Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061,China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2023年第1期65-76,共12页
Journal of Beijing University of Traditional Chinese Medicine
基金
湖北省第二届医学领军人才工程培养对象暨湖北名医工作室负责人项目(No.鄂卫通[2019]47号)
湖北省卫生计生委中西医结合重点科研项目(No.鄂卫生计生通[2017]20号)
湖北省卫生健康委员会中医药科研项目青年人才项目(No.ZY2021Q031)
湖北省卫健委面上重点项目(No.WJ2019H422)。
关键词
脑衰老
首乌丸
病理形态
肠道菌群
氧化三甲胺
大鼠
brain aging
Shouwu Pill
pathological morphology
intestinal flora
trimethylamine oxide
rats
作者简介
张超,女,在读博士生;通信作者:陈刚,男,博士,教授,博士生导师,主要研究方向:中医药延缓衰老研究,E-mail:250149875@qq.com。
