摘要
目的:基于16S rRNA技术研究黄连酒蒸前后对正常和2型糖尿病大鼠肠道菌群的影响,从肠道菌群的角度探讨黄连酒蒸炮制'减毒增效'的原理。方法:采用高脂饮食结合链脲佐菌素复制2型糖尿病大鼠模型。正常和模型大鼠分别按0. 8 g·kg-1(按生药量计)每天灌胃生黄连生品或酒蒸黄连水煎液,阳性药组按0. 25 g·kg-1每天灌胃二甲双胍溶液,连续给药30 d。每周测1次血糖,于给药第27天进行口服糖耐量的测定。于第30天取粪便后麻醉大鼠,取血进行酶联免疫吸附试验(ELISA)检测,解剖取结肠进行苏木精-伊红(HE)染色。基于16S rRNA基因测序技术对各个组大鼠肠道菌群的alpha多样性,beta多样性,菌群丰度及构成,及其与抗炎抗氧化的相关性等进行综合评价。结果:正常大鼠给予黄连生品或酒蒸黄连后,出现了一定的炎性反应,氧化-抗氧化系统失衡,结肠病理性损伤以及肠道菌群改变,表现出了一定的毒副作用,但酒蒸黄连的毒副作用低于生黄连。黄连酒蒸前后均能降低2型糖尿病大鼠的空腹血糖以及提高糖耐量,虽然酒蒸后作用有增强的趋势,但酒蒸前后无显著差异。2型糖尿病大鼠出现轻度的炎症反应和氧化-抗氧化系统失衡,表现为白细胞介素(IL)-6,核转录因子(NF)-κB和丙二醛(MDA)含量显著升高(P <0. 05,P <0. 01),谷胱甘肽过氧化物酶(GSH-Px),还原型谷胱甘肽(GSH),超氧化物歧化酶(SOD),过氧化氢酶(CAT)活力明显降低(P <0. 01),黄连酒蒸前后均能抑制炎症反应以及氧化-抗氧化系统失衡,且酒蒸黄连作用优于黄连生品;2型糖尿病大鼠出现明显的结肠病理性损伤,而黄连酒蒸前后均能减轻这种改变,且酒蒸黄连作用优于生黄连。2型糖尿病大鼠出现明显的肠道菌群紊乱,包括下调拟杆菌门(Bacteroidetes),厚壁菌门(Firmicutes)和Kiritimatiellaeota,上调变形菌门(Proteobacteria),放线菌门(Actinobacteria),酸杆菌门(Acidobacteria),�
Objective:To investigate the effect of Coptidis Rhizoma on intestinal microflora of normal and type 2 diabetic rats before and after being steamed with rice wine based on 16S rRNA sequencing technology.Method:The type 2 diabetic rat model was established by high-fat diet and streptozotocin.Normal and model rats were given the decoction of Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine every day(0.8 g·kg-1,according to raw drug quantity),the positive drug group was given metformin solution every day(0.25 g·kg-1),continuous administration for 30 d.Blood glucose was measured once a week and oral glucose tolerance was measured on the 27 thday of administration.On the 30 thday,feces were taken and the rats were anesthetized.Blood was taken for enzyme linked immunosorbent assay(ELISA).Colon was dissected for hematoxylin-eosin(HE)staining.Based on 16 S rRNA gene sequencing technology,the alpha diversity,beta diversity,abundance and composition,and the correlation with anti-inflammatory and antioxidant activities of intestinal microflora were comprehensively analyzed and evaluated.Result:After giving normal rats Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine,there were some inflammatory reactions,imbalance of oxidation-antioxidant system,pathological damage of colon and changes of intestinal microflora,which showed some toxic and side effects,but the toxic and side effects of Coptidis Rhizoma steamed with rice wine were lower than those of Coptidis Rhizoma.Coptidis Rhizoma could reduce fasting blood glucose and improve glucose tolerance in type 2 diabetic rats before and after steaming.Although the effect of Coptidis Rhizoma steamed with rice wine had an increasing trend,there was no significant difference between before and after processing.Type 2 diabetic rats appeared mild inflammation and imbalance of oxidation-antioxidant system,characterized by significant increase of contents of interleukin(IL)-6,nuclear transcription factor(NF)-κB and malondialdehyde(MDA)(P<0.05,P<0.01),significant de
作者
王瑾
冉倩
王琳
胡昌江
黄勤挽
张海
WANG Jin;RAN Qian;WANG Lin;HU Chang-jiang;HUANG Qin-wan;ZHANG Hai(Key Laboratory of Technology of Chinese Medicine Processing,National Administration of Traditional Chinese Medicine(TCM),School of Pharmacy,Chengdu University of TCM,Chengdu 611137,China;Ya'an Xunkang Pharmaceutical Co.Ltd.,Ya'an 625600,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第22期92-102,共11页
Chinese Journal of Experimental Traditional Medical Formulae
基金
四川省中医药管理局中医药基础研究项目(2018JC011)
成都市科技局科技惠民技术研发项目(2015-HM01-00401-SF)
关键词
黄连
生品
酒蒸品
16S
rRNA技术
2型糖尿病
肠道微生物
抗氧化活性
Coptidis Rhizoma
raw products
steamed products
16S rRNA technique
type 2 diabetes mellitus
intestinal microflora
antioxidant activity
