不同剂量槲皮素干预对菌群失衡小鼠肠道微生态改善作用  

QUERCETIN INTERVENTION IMPROVES GUT MICRO-ENVIRONMENT IN MICE TREATED WITH ANTIBIOTICS A DOSE-RESPONSE STUDY

作　　者：石塔拉 吕鹏[1] 胡志勇 练武[1] 宓伟[1] SHI Ta-la;LU Peng;HU Zhi-yong;LIAN Wu;MI Wei(Shool of Public Health,Binzhou Medical University,Yantai 264003,China)

机构地区：[1]滨州医学院公共卫生学院,烟台264003

出　　处：《营养学报》2024年第4期356-364,共9页Acta Nutrimenta Sinica

基　　金：国家自然科学基金项目(No.81602842);山东省医药卫生科技发展计划(No.202012020832)。

摘　　要：目的比较不同剂量槲皮素对小鼠肠道微环境的影响,探讨槲皮素与肠道菌群相互作用的有效浓度。方法随机将60只雄性昆明小鼠分成对照组(control,C),模型组(model,M,0 mg/kg槲皮素)、低剂量槲皮素组(low quercetin,LQ,200 mg/kg),中剂量槲皮素组(moderate quercetin,MQ,400 mg/kg)、高剂量槲皮素组(high quercetin,HQ,600 mg/kg)。C组生理盐水灌胃,M和LQ,MQ,HQ组小鼠用抗生素混合液灌胃7d,通过16S扩增子测序结果中菌群多样性比较来确定是否建立肠道菌群失衡模型。建模成功的小鼠用含不同剂量的槲皮素饲料饲养10d并收集粪便、组织、血液样品。用16S rDNA高通量测序分析小鼠肠道微生物结构、气相色谱仪测定粪便短链脂肪酸含量、HE染色分析小肠组织变化、ELISA分析肠组织通透性指标。结果与C组比较,7 d的抗生素灌胃有效清除了肠道微生物(除变形菌门、厚壁菌门和拟杆菌门),成功建立了肠道菌群失衡小鼠模型。槲皮素干预后与M组比较,LQ、MQ、HQ可降低厚壁菌门/拟杆菌门的比例,分别降低了32.8%、66.3%、62.9%。在属水平,三种剂量槲皮素对Escherichia-Shigella和Klebsiella等条件致病菌属的抑制率均达到99%以上。LQ组中增加的Lactococcus和Desulfovibrio菌属与肠道菌群代谢产物丁酸含量增加有较强的正相关性(相关系数分别是0.74和0.78,P<0.001),LQ组粪便丁酸含量[(0.27±0.12)mmol/kg]与M组[(0.04±0.02)mmol/kg]比较显著增加(P<0.05),但MQ和HQ组丁酸含量无显著差异。此外,HE染色结果显示,与M组比较,随槲皮素剂量的增加肠组织黏膜和绒毛的恢复越明显,血清中二胺氧化酶含量随槲皮素剂量的增加分别降低29.25%,37.90%,41.49%,作用有剂量依赖性。结论低剂量槲皮素有利于产丁酸菌属等肠道有益菌属的繁殖,中、高剂量槲皮素则通过竞争性抑制条件致病菌,修复肠组织结构,稳定肠微生态环境。Objective To investigate the effects of different doses of quercetin on gut microbiota in mice treated with antibiotics,and explore effective dose of quercetin.Methods Sixty male Kunming mice were randomly divided into control group(C),model group(M,0 mg/kg quercetin),low dose quercetin group(LQ,200 mg/kg),moderate dose quercetin group(MQ,400 mg/kg),and high dose quercetin group(HQ,600 mg/kg).The gut microbial disorder was induced by gavage of an antibiotic cocktail.The C and M groups were fed the AIN-93G diet,and LQ,MQ,HQ groups were fed the AIN-93G diets with different doses of quercetin for 10 days.At the end of experiment,the fecal,small intestinal tissue,blood samples were collected.16SrDNA high-throughput sequencing,gas chromatography,hematoxylin-eosin staining and ELISA methods were used to analyze gut microbial structure,short chain fatty acids,intestinal tissue and gut permeability,respectively.Results Compared with the C group at phylum level,7 days antibiotic cocktail treatment decreased gut microbial diversity,except Proteobacteria,Firmicutes,and Bacteroidetes,which indicate gut microbial disrupted model were successfully established.Compared with the M group,LQ,MQ,HQ groups decreased Firmicutes/Bacteroides ratio by 32.8%、66.3%、62.9%,respectively.At the genus level,the inhibitory rates of different doses of quercetin on Escherichia Shigella and Klebsiella were all up to 99%.The increased bacteria genus Lactococcus and Desulfovibrio in the LQ group were positively related with butyric acid content(correlation coefficients were 0.74 and 0.78,respectively,P<0.001).The fecal butyric acid content in the LQ group((0.27±0.12)mmol/kg)was significantly higher than in the M group((0.04±0.02)mmol/kg,P<0.05).In addition,the recovery of intestinal mucosa and villi became more pronounced with the increase of quercetin dosage.The content of diamine oxidase in serum decreased by 29.25%,37.90%,and 41.49%,respectively.Conclusion Lower quercetin dose was beneficial for butyric acid producing bacteria growth,while

关 键 词：槲皮素 肠道菌群 短链脂肪酸 生物利用 

分 类 号：R151.1[医药卫生—营养与食品卫生学]