基于16S rRNA测序探究葛根芩连汤对抗生素相关性腹泻肠道菌群结构的影响

16S rRNA Sequencing-based Investigation of the Effects of Gegen Qinlian Decoction on Intestinal Flora Structure in Antibiotic-associated Diarrhea

应用16S rRNA测序方法,观察葛根芩连汤对抗生素相关性腹泻SD大鼠肠道菌群结构的影响;将60只SD大鼠适应性喂养1周后,随机分为2组分别为空白组(C)(10只)和造模组(50只),其中空白组给予生理盐水灌胃,而造模组SD大鼠给予克林霉素250 mg/kg灌胃造模,两组均为1次/d,连续7 d。造模成功后,将其随机分为5组:丽珠肠乐组(P)、模型组(M)、葛根芩连汤高、中、低剂量组(GQD-H、GQD-M、GQD-L),每组10只。丽珠肠乐组(双歧杆菌活性菌胶囊,0.15 g/kg)、葛根芩连汤高剂量组(10.08 g/kg)、中剂量组(5.04 g/kg)低剂量组(2.52 g/kg)分别给予对应药物进行灌胃干预。空白组及模型组给予等体积生理盐水灌胃,1次/d。各组连续给药7 d后,在无菌操作下提取各组大鼠粪便DNA,进行16S rRNA测序并分析其结果。通过Alpha与Beta多样性分析发现,GQD具有增加抗生素相关性腹泻大鼠肠道菌群多样性的作用;GQD可在门水平增加厚壁菌门、拟杆菌门的丰度,在属水平增加乳酸杆菌属、拟杆菌属的丰度。通过LEfse分析,发现GQD能增加乳酸杆菌、拟杆菌丰度,降低变形菌丰度。提示GQD可能通过改善肠道微生态结构治疗抗生素相关性腹泻。

The effects of Gegen Qinlian decoction(GQD)on intestinal flora structure in SD rats with antibiotic-associated diarrhea(AAD)were investigated using 16S rRNA sequencing.First,60 SD rats were subjected to 1 week of adaptive feeding,after which they were randomly divided into two groups:the blank control(C)group of 10 rats and the model construction group of 50 rats.The model construction group was administered 250 mg/(kg·day)clindamycin by gavage for 7 consecutive days for model establishment,whereas the blank control group was administered saline by gavage once daily during the same period.After successful model construction,the model construction group was randomly divided into five groups of 10 rats each:the model(M),live bifidobacterium capsule(P),high-dose GQD(GQD-H),medium-dose GQD(GQD-M),and low-dose GQD(GQD-L)groups.The P group was administered 0.15 g/kg of bifidobacteria and the GQD-H,GQD-M,and GQD-L groups were respectively administered 10.08,5.04,and 2.52 g/kg GQD by gavage once daily for 7 consecutive days;the C and M groups were administered equivalent volumes of saline by gavage once daily during the same period.After treatment,fecal DNA was extracted from each group aseptically and subjected to 16S rRNA sequencing.Alpha and beta diversity analysis showed that GQD increased the diversity of intestinal flora in rats with AAD.GQD increased Firmicutes and Bacteroides abundance at the phylum level and Lactobacillus and Bacteroides abundance at the genus level.LEfSe analysis revealed that GQD increased Lactobacillus and Bacteroides abundance but reduced Proteobacteria abundance.These results suggest that GQD may effectively treat AAD through the improvement of intestinal microecological structure.