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1.天津中医药大学 现代中药发现与制剂技术教育部工程研究中心,天津 301617
2.天津中医药大学 方剂学教育部重点实验室,天津 301617
苗小雨,在读硕士,从事药物制剂与新药开发研究,E-mail:mmiaoxiaoyu@163.com
王亚静,硕士,研究员,从事药物制剂与新药开发研究,Tel:022-59596169,E-mail:yajing022@163.com; *
赵鑫,博士,副研究员,从事中药微生物组学研究,E-mail:x.zhao26@tjutcm.edu.cn
收稿日期:2022-07-27,
网络出版日期:2022-10-21,
纸质出版日期:2023-02-20
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苗小雨,王亚静,赵鑫等.乌梅散对菌群失调性腹泻小鼠肠道乳糖酶活性及菌群多样性的影响[J].中国实验方剂学杂志,2023,29(04):33-42.
MIAO Xiaoyu,WANG Yajing,ZHAO Xin,et al.Effect of Wumeisan on Gut Lactase Activity and Microflora Diversity of Mice with Dysbacteriosis Diarrhea[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(04):33-42.
苗小雨,王亚静,赵鑫等.乌梅散对菌群失调性腹泻小鼠肠道乳糖酶活性及菌群多样性的影响[J].中国实验方剂学杂志,2023,29(04):33-42. DOI: 10.13422/j.cnki.syfjx.20221906.
MIAO Xiaoyu,WANG Yajing,ZHAO Xin,et al.Effect of Wumeisan on Gut Lactase Activity and Microflora Diversity of Mice with Dysbacteriosis Diarrhea[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(04):33-42. DOI: 10.13422/j.cnki.syfjx.20221906.
目的
2
探究乌梅散对菌群失调性腹泻小鼠的乳糖酶活性和肠道菌群多样性的影响。
方法
2
将小鼠随机分为4组,分别为正常组、模型组及乌梅散低、高剂量组,每组8只。采用混合抗生素灌胃造模7 d,乌梅散低、高剂量组灌胃乌梅散(5.98、11.96 g·kg
-1
)连续给药7 d,正常组与模型组给予等体积无菌水。记录小鼠腹泻情况、体质量、摄食量的变化,末次给药后收集粪便采用比色法测定乳糖酶活性,并运用16S rRNA高通量测序技术检测并分析肠道菌群的变化。
结果
2
与正常组比较,模型组小鼠大便稀软,体质量减轻、摄食量减少,乳糖酶活性降低,肠道菌群多样性明显降低,菌群相对丰度门、属水平均发生明显变化。与模型组比较,乌梅散降低了小鼠的腹泻率并促使体质量及摄食量快速回升,升高了抗生素所降低的乳糖酶活性,提高菌群失调小鼠的群落丰富度、多样性,并促使物种组成向正常组靠近;乌梅散给药组回调了菌群失调性腹泻小鼠的3种差异菌门(拟杆菌门、变形菌门、疣微菌门)与9种差异菌属(臭气杆菌属、肠杆菌属、无害梭菌属等),其中
norank f Muribaculaceae
属、
norank f norank o Clostridia UCG-014
属、
Lachnospiraceae NK4A136 group
属、臭气杆菌属与体质量、乳糖酶活性呈显著正相关,大肠埃希菌-志贺菌属、肠杆菌属、肠球菌属、无害梭菌属与腹泻率呈显著正相关。功能预测发现,高剂量乌梅散能明显回调富集过高的新陈代谢、遗传信息处理、人类疾病等6个功能水平,对内分泌代谢病、免疫性疾病、寄生虫传染性疾病等有积极影响。
结论
2
乌梅散能缓解菌群失调性腹泻的症状,其机制可能是通过提高乳糖酶活性、调节肠道菌群结构而实现。
Objective
2
To explore the effects of Wumeisan on gut lactase activity and microflora diversity of mice with dysbacteriosis diarrhea.
Method
2
The mice were randomly divided into 4 groups, namely, the normal group, the model group, and the low-dose and high-dose Wumeisan groups, with 8 mice in each group. The mouse model was made by gavage of mixed antibiotics for 7 d, and the low-dose and high-dose Wumeisan groups (5.98, 11.96 g·kg
-1
) were given gavage for 7 d continuously. The normal group and the model group were given the same volume of sterile water. The changes in the body weight, food intake, and diarrhea of mice were recorded. Feces were collected after the last administration, and the lactase activity was detected by the colorimetric method. The gut microbiota changes were detected by the 16S rRNA high-throughput sequencing technology.
Result
2
Compared with those in the normal group, the mice in the model group had dilute and soft stools, reduced body mass, reduced food intake, reduced lactase activity, significantly reduced intestinal flora diversity, and significant changes in the relative abundance phylum and genus levels of flora. Compared with the model group, Wumeisan reduced the diarrhea rate of mice, promoted the rapid recovery of body weight and food intake, increased the lactase activity decreased by antibiotic, improved the community abundance and diversity of mice with dysbacteriosis, and made the species composition closer to that in the normal group. The abundance of three phyla (Bacteroidota, Proteobacteria, Verrucomicrobiota) and nine genera (
Odoribacter
,
Enterococcus
,
Clostridium innocuum group
,
etc.) of mice with diarrhea were regulated by Wumeisan. Among them,
norank f Muribaculaceae
,
norank f norank o Clostridia UCG-014
,
Lachnospiraceae NK4A136 group
, and
Odoribacter
showed significant positive correlation with the body weight and lactase activity, and
Escherichia-Shigella
,
Enterobacter
,
Enterococcus
, and
Clostridium innocuum group
showed significant positive correlation with the diarrhea rate. Function prediction showed that the high-dose Wumeisan significantly reseted 6 functional levels of metabolism, genetic information processing, and human diseases, and had positive effects on endocrine and metabolic diseases, immune diseases, infectious disease, and parasitic infectious diseases.
Conclusion
2
Wumeisan can relieve the symptoms of dysbacteriological diarrhea by increasing the lactase activity and regulating the gut microbiota composition.
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