Dextran sulfate sodium (dss)

Six-to-eight-week-old female C57BL/6 (wild-type, WT) mice were obtained from the Hunan Silaike Jingda Laboratory Animal Co., Ltd., (Changsha, China), and miR-10b knockout (KO) mice (generated on C57BL/6 background) were purchased from Nanjing Biomedical Research Institute of Nanjing University (Nanjing, China). All mice were fed autoclaved food and water and maintained in a specific-pathogen-free facility at Hunan Normal University (Changsha, China). To induce colitis, WT and miR-10b KO mice were given drinking water containing 4% DSS (MW 36,000–50,000; Meilunbio, Dalian, China) for 7 consecutive days. Mice in control groups were given normal drinking water only. Survival rates, body weight loss, rectal bleeding, and stool consistency were monitored daily. The disease activity index (DAI) scores were the sum of body weight loss, fecal hardness, and bleeding scores as previously described (Supplementary Table S1) [16 (link)]. After 7 days of DSS treatment, mice were euthanized by CO₂ inhalation. Serum and colon samples were harvested and stored in a refrigerator at −80 °C for subsequent experiments.

Age- and sex-matched control and experimental mice were administered 3% DSS (Meilun Biotechnology Co., Ltd., China) in drinking water for 7 days. Body weight was monitored every 24 h. On day 7, mice were anesthetized and sacrificed and the length of the entire colon was measured. Colitis was assessed using hematoxylin and eosin staining, and severity scores were calculated. Histological evaluation of DSS-induced colitis. The evaluation of the histological changes in the colon was performed according to the published criteria (35 ). The scoring system was put in Supplementary Table 2.

DSS (purity >98%; molecular weight 198.17), purchased from Meilunbio (Dalian, China), was dissolved in sterile distilled water as a 40-mM stock solution. NIH3T3 cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in Dulbecco’s modified eagle medium (DMEM, Gbico, USA) containing 10% fetal bovine serum (FBS) in a 37°C, 5% CO₂ incubator. Then, NIH3T3 cells were treated in different ways as followed: part of NIH3T3 cells were treated with DSS at different concentrations (0, 25, 50, 100, 200, and 400 μM) for 24 h; part of NIH3T3 cells were pretreated with various concentrations of DSS (0, 25, 50, 100, 200, and 400 μM) for 2 h, followed by stimulation with or without TGF-β1 (5 ng/mL) for 24 h and another part of NIH3T3 cells were pretreated with DSS (50 μM) for 2 h, followed by the stimulation with or without TGF-β1 (5 ng/mL) for 24 h.

PCA (>98%) was purchased from MACLIN (Shanghai, China). DSS was purchased from Meilunbio (Dalian, China). Occludin (GB111401) antibody was purchased from Servicebio, Inc. (Wuhan, China). GPX4 (AF301385) antibody was purchased from AiFang biological, Inc. (Changsha, China). HRP-conjugated secondary antibody (GAR007-100) was purchased from MultiSciences, Inc. (Wuhan, China). RIPA Lysis Buffer (CW2333) was purchased from CWBIO (Taizhou, China). Tumor necrosis factor-alpha (TNF-α) (70-EK282/3-96) and interleukin-6 (IL-6) (70-EK206/3-96) enzyme-linked immunosorbent assay (ELISA) kits were purchased from MultiSciences Inc. (Hangzhou, China). An interleukin-12 (IL-12) (E-MSEL-M0004) ELISA kit was purchased from Elabscience, Inc. (Wuhan, China). MDA (A003-1) and GSH (A006-2-1) kits were purchased from Nanjing Jiancheng Bioengineering Institute, Inc. (Nanjing, China). An iron assay kit was purchased from Solarbio Life Sciences Institute, Inc. (Beijing, China). An ROS Assay kit (S0033) was purchased from Beyotime Biotechnology Institute, Inc. (Shanghai, China). A BCA detection kit was purchased from NCM Biotechnology, Inc. (NCM Biotech, Suzhou, China), and a CCK-8 kit (C0037) was purchased from Beyotime Biotechnology Institute, Inc. (Shanghai, China).

DSS was purchased from Meilun Biotechnology Co., Ltd. (Dalian, China). Mesalazine was purchased from Losan Pharma GmbH (Freiburg im Breisgau, Germany). Artemisinin was purchased from TCI Development Co., Ltd. (Shanghai, China). ELISA kits for IL-1β, IL-13, IL-33, and ST2 were purchased from Jiangsu Enzyme Immune Industrial Co., Ltd. (Jiangsu, China). ELISA kit for IL-17 was purchased from Shanghai Senxiong Technology Industry Co., Ltd. (Shanghai, China). ELISA kit for IL-23 was purchased from Shanghai XiTang Biological Technology Co., Ltd., (Shanghai, China).