Gut microbiota modulated by CiLi (Rosa roxburghii Tratt.) polyphenols improves colitis through microbe-lipid mediator-immunity axis

Sample collection
Five-week-old male C57BL/6J mice were purchased from Beijing Vital River Laboratory Animal Science and Technology Co. (Beijing, China) and kept in a standard facility for specific pathogen-free animals with a light and dark cycle of 12 h. Animal experiments were conducted in accordance with the protocols approved by the Animal Care and Ethics Committee of China Agricultural University (No. AW32602202-4-2).

Treatment of DSS-induced colitis mice with CL_PP
CL_PP (100 mg/kg) were orally administered for 4 weeks, and 2.5% DSS was added to the drinking water for 7 days to induce acute colitis during the last week. Body weight and DAI were measured daily during the DSS treatment. Blood was collected from the eyeball in EDTA anticoagulant tubes and centrifuged to obtain plasma.

Treatment of antibiotic-treated mice with CL_PP
The mice were given a mixture of antibiotic cocktail (100 mg/L neomycin, 50 mg/L streptomycin, 100 mg/L penicillin, 50 mg/L vancomycin and 100 mg/L metronidazole) in drinking water during the whole experiment. After 2 weeks, the mice were divided into two groups, and CL_PP or PBS was orally administered every day for 3 weeks. Colitis was induced by treatment with 2.5% DSS in the drinking water for 7 days.

Treatment of DSS-induced colitis mice with CL_PP_FMT
Mice were divided into 4 groups (the PBS, PBS_FMT, CL_PP, and CL_PP_FMT groups). The two groups (the PBS_FMT and CL_PP_FMT groups) were treated with the antibiotic cocktail mentioned above for 2 weeks. The feces of mice in the PBS and CL_PP groups were collected, and the homogenate was diluted, filtered through a 70 μm filter and subsequently subjected to FMT. The final concentration was 50 mg of feces/mL. After 2 weeks of fecal treatment, 2.5% DSS solution was added to the drinking water of the mice in the PBS_FMT group and CL_PP_FMT group to induce colitis.

样本收集
五周大的雄性C57BL/6J小鼠购自北京维通利华实验动物科学技术有限公司（北京，中国），并在具有12小时光暗周期的标准无特定病原体动物设施中饲养。动物实验遵循中国农业大学动物护理与伦理委员会批准的方案（批准号：AW32602202-4-2）。

柳氮磺吡啶诱导的结肠炎小鼠的CL_PP治疗
以CL_PP（100 mg/kg）进行口服给药4周，并在实验最后一周将2.5%的DSS添加至饮用水中，以诱导急性结肠炎。在DSS治疗期间，每日测量体重和DAI。使用含有EDTA抗凝剂的试管从眼球中采集血液，并进行离心以获得血浆。

抗生素治疗小鼠的CL_PP处理
在整个实验过程中，小鼠饮用含有抗生素混合物（新霉素100 mg/L，链霉素50 mg/L，青霉素100 mg/L，万古霉素50 mg/L和甲硝唑100 mg/L）的饮用水。经过2周的抗生素处理后，将小鼠分为两组，每组每天口服CL_PP或PBS，持续3周。通过在饮用水中添加2.5%的DSS，持续7天来诱导结肠炎。

DSS诱导的结肠炎小鼠的CL_PP_FMT治疗
将小鼠分为四组（PBS组、PBS_FMT组、CL_PP组和CL_PP_FMT组）。其中两组（PBS_FMT组和CL_PP_FMT组）接受了上述抗生素混合物的治疗，持续2周。收集PBS组和CL_PP组的粪便，进行均质化处理，稀释后通过70 μm的滤器过滤，随后进行粪便微生物移植（FMT）。最终粪便浓度为50 mg粪便/mL。在粪便治疗2周后，向PBS_FMT组和CL_PP_FMT组的饮用水中添加2.5%的DSS溶液，以诱导结肠炎。