Data Availability StatementAll datasets generated because of this study are included in the article/supplementary material. PD-ligand 1 on the surface of Tfh cells. SSP inhibited activation of BcL-6, phosphorylated signal transducer and activator of transcription (p-STAT)3, signal lymphocyte activation molecule (SLAM)-associated protein but improved Blimp-1 and STAT3 expression in colonic tissues. The results indicated that SSP regulated the differentiation and function of Tfh cells to treat IBD, which was potentially related with inhibiting the Bcl-6/Blimp-1 pathway. (Juss.) Benth., (Turcz.) Baill, L., Houtt., Mill., and Rosc. Which were prepared into pills according to the dose ratio (100, 200, 400, 200, 200, and 200 g, ratio: 1:2:4:2:2:2, respectively). SSP contained deoxyschizandrin (72.6 g/g), -schizandrin (131.5 g/g), schizandrin (258.0 g/g), schizandrol B (71.2 g/g), schisantherin A (25.1 g/g), psoralen (131.08 g/g), isopsoralen (1293.7 g/g), evodiamine (22.2 g/g), and rutaecarpine (24.0 g/g). SSP was analyzed by high-performance liquid chromatography coupled with electrospray tandem mass spectrometry (Zhang et al., 2018). DSS (molecular weight: 36,000C50,000 kDa; No. 160110) was obtained from MP Biomedicals (Santa Ana, CA, USA). Mesalazine was obtained from Jiamusi Luling Pharmaceutical Co., Ltd., Company (Jiamusi, China). Animals Male BALB/C mice weighing 18C22 g (Animal Certificate No. SCXK 2006-0008) were purchased from Hunan Slake Jing da Experimental Animal Co., Ltd. (Changsha, China). All animals were housed in specific-pathogen-free conditions, with standard laboratory diet, 12-h light/dark cycle and constant room temperature, and had free access to standard diet and tap water according to the guidelines of the Animal Center. This Protocol (License No.: JZ2018-105) was approved by the Institutional Animal Care and Use Committee (IACUC) of Jiangxi University of Traditional Chinese Medicine. The mice were acclimated for 3 days according to the IACUC Animal Welfare Guidelines before formal experiments were performed. Thirty-two mice were divided into two groups: eight mice were in the normal group and the remaining mice were treated by DSS to induce colitis. The twenty-four mice were observed to have bloody stools around the fourth or fifth day after DSS treatment, which hinted the colitis was successfully induced. Twenty-four colitis mice were randomly divided into three groups: DSS: colitis without treatment; DSS + SSP: colitis treated with SSP; and DSS + 5-ASA: colitis treated with 5-aminosalicylic acid (ASA). Eight colitis mice were in every group. DSS-Induced Colitis According to the previous study (Yoshihara et al., 2006), colitis was induced PSI-7976 in BALB/C mice with 3% (w/v) DSS (molecular weight: 36,000C50,000 kDa) dissolved in deionized water drunk ad libitum (days 1C7). Fresh 3% DSS solutions were made every PSI-7976 morning in deionized water. Control mice were given tap water. Therapeutic Protocols On day 8, the DSS + SSP group was administered 2.5 g kg?1 SSP dissolved in physiological saline by oral gavage for 7 Rabbit Polyclonal to NDUFS5 days; the DSS + 5-ASA group was administered 300 mg kg?1 mesalazine by PSI-7976 oral gavage for 7 days; and mice in DSS and normal groups were treated with the same volume of physiological saline by oral gavage for 7 days. On day 15, all animals were sacrificed under sodium pentobarbital (50 mg/kg ip) anesthesia. Macroscopic Evaluation The mice were weighed before anesthesia, and the colons were quickly removed. The colon length and weight were measured, and the colon weight index (CWI), CWI = colon weight/body weight 100 was calculated. Hematoxylin and Eosin (H&E) Staining and Microscopic Evaluation The colon was preserved in a 4% polyformaldehyde solution for 7 days, then dehydrated and embedded in paraffin, and the paraffin sections were serially sectioned at 4 m. The tissue sections were PSI-7976 dewaxed and rehydrated using an alcohol gradient and stained with H&E. The pathological features of the colon were observed and evaluated under a microscope. The histological grading of colitis was as described by Dieleman et al. (1998). Inflammation: none, 0 points; slight, 1 point; moderate, 2 points; and severe, 3 factors. Extent: non-e, 0 factors; mucosa, 1 stage; and submucosa and mucosa, 3 factors. Regeneration: no tissues repair, 4 factors; surface epithelium not PSI-7976 really intact, 3 factors; regeneration with crypt depletion,.