Thioguanine effects on mice gut microbiota. Thioguanine and microbiota

Male and or female mice were intra-gastrically gavaged daily with either vehicle control or thiopurine drugs (TG or MP). Caecal content (CC), caecal mucosa (CM) and faeces (F) of WT, Hprt-/- and Winnie mice were collected at day 14 post-TG administration at doses of 0, 0.5 mg/kg or 2.5 mg/kg (Winnie only). DNA was extracted from faeces at day 0 and day 14; CM and CC at day 14. DNA extracts were used as the template for PCR-based amplification of the bacterial V1-V3 region of the 16S rRNA gene using a barcoded pyrotagging approach, and sequenced with a Roche 454 FLX+ with titanium chemistry.

将雌雄小鼠每日经胃灌胃给予溶媒对照或硫嘌呤类药物（TG或MP）。于TG给药后第14天，采集野生型（WT）、Hprt基因敲除小鼠（Hprt-/-）及Winnie小鼠的盲肠内容物（CC）、盲肠黏膜（CM）与粪便（F）；其中仅Winnie小鼠的给药剂量设为0、0.5 mg/kg或2.5 mg/kg，其余基因型小鼠给药剂量为0。分别于第0天和第14天采集粪便样本提取DNA，于第14天采集盲肠黏膜与盲肠内容物样本提取DNA。以提取的DNA为模板，采用带条形码的焦磷酸标记策略，通过聚合酶链式反应（PCR）扩增细菌16S rRNA基因的V1-V3可变区，随后使用搭载钛试剂化学的罗氏454 FLX+测序平台进行测序。