ABSTRACT
This study aims to investigate the regulatory effect of Sishen Pills(SSP) and its split prescriptions Ershen Pills(EP) and Wuweizi Powder(WP) on T follicular helper(Tfh) cell subset in the dextran sodium sulfate(DSS)-induced colitis mice and the mechanism. A total of 60 male SPF BALB/c mice were used, 10 of which were randomly selected as the normal group. The rest 50 were induced with 3% DSS solution for colitis modeling. After modeling, they were randomized into 5 groups: model group, SSP group, EP group, WP group, and mesalazine group. Body mass, colon mass, colon mass index, colon length, and unit colon mass index in each group were observed. After hematoxylin-eosin(HE) staining, the pathological injury of colon tissue was scored. The expression levels of molecules related to the STAT/SOCS signaling pathway in colon tissues were analyzed by Western blot. Differentiation levels of Tfh cells such as CD4~+CXCR5~+IL-9~+(Tfh9), CD4~+CXCR5~+IL-17~+(Tfh17), and CD4~+CXCR5~+Foxp3~+(Tfr) in peripheral blood of mice were detected by flow cytometry. The results showed each treatment group demonstrated significant increase in body mass and colon length, decrease in colon mass, colon mass index, unit colon mass index, and histopathological score(P<0.05, P<0.01), reduction of the expression of p-STAT3, STAT3, p-STAT6, and STAT6(P<0.05, P<0.01), rise of the expression of SOCS1 and SOCS3(P<0.05, P<0.01), decrease of Tfh9 and Tfh17 cells, and increase of Tfr cells(P<0.05, P<0.01) compared with the model group. These results indicated that SSP and the split EP and WP may alleviate ulcerative colitis by inhibiting the activation of STAT/SOCS signaling pathway and regulating the balance of Tfr/Tfh9/Tfh17 cells.
Subject(s)
Animals , Male , Mice , Colitis/genetics , Colitis, Ulcerative/metabolism , Mice, Inbred BALB C , Prescriptions , T-Lymphocytes, RegulatoryABSTRACT
Objective:To observe the effect of Wendantang on tumor necrosis factor-α (TNF-α),interleukin(IL)-6,IL-17,IL-22 and other related inflammatory factors in peripheral blood and the expression of STAT3[the key molecule of Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) signal pathway in hypothalamus] mRNA and protein of obese rats with syndrome of phlegm-dampness,so as to explore the internal mechanism of Wendantang in interfering obesity with syndrome of phlegm-dampness. Method:A total of 100 rats were randomly divided into blank group(30 rats) and modeling group(70 rats),rats in the blank group was fed with basic feed and the modeling group was fed with high-fat feed for 6 weeks.Animal model of obesity with syndrome of phlegm-dampness was established by the method in literature.After successful modeling,16 obese rats were selected and randomly divided into the model group and Wendantang intervention group with 8 rats in each group,and another 8 rats in the blank group were randomly selected as the normal group.Rats in Wendantang intervention group were given 15 g·kg-1 of crude drug by gavage,while the model group and the normal group were given the same amount of distilled water for gavage once a day for 6 weeks.No eating but no prohibiting drinking before dealing with 12 h and then taking samples after anesthesia.The body weight,Lee's index and obesity rate of rats were measured,the levels of blood lipids[total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C) and high density lipoprotein cholesterol(HDL-C)] of rats were detected with a full-automatic biochemical analyzer according to the requirements of the kit,the expression of TNF-α,IL-17,IL-22 and IL-6 in peripheral blood of rats was detected with enzyme-linked immunosorbent assay(ELISA),STAT3 mRNA expression in hypothalamus of rats was detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and the expression of STAT3 protein in hypothalamus of rats was detected with Western blot. Result:The high-fat feed feeding could successfully replicate the obese rat model,and the obesity rate of rats in the modeling group was greater than 20%,and compared with the blank group,the body weight and Lee's index of rats in the modeling group were significantly increased(PPPPPPPPPPPConclusion:Wendantang has a good effect on improving phlegm-dampness in obese rats,and the mechanism may be related to regulating JAK2/STAT3 signal pathway then to improve the chronic inflammatory state of the body,and all of which provides a scientific basis for Wendantang in intervening obesity with syndrome of phlegm-dampness.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of scorpion and centipede on interleukin (IL)-2, IL-4, IL-10 in the small intestinal mucosa and joint injury of rats with collagen induced arthritis (CIA).</p><p><b>METHODS</b>Sixty Wistar rats were randomly divided into 6 groups: the normal control group, the model group, the low dose scorpion and centipede group, the middle dose scorpion and centipede group, the high dose scorpion and centipede group, and the type II collagen treatment group. The joints' volume was measured 40 days after type II collagen (CII) induced rheumatoid arthritis model was established. The joint injury was observed by naked eyes. The expression levels of IL-2, IL-4, and IL-10 in the small intestine tissue homogenate were detected by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The joint injury score and volume of two hind limbs were obviously higher in the model group than in the normal control group since the 23rd day (P < 0.01). Rats were accompanied with red, swollen, and deformed foot toes and ankle joints. Walking was even affected. Meanwhile, the joint injury score and volume of two hind limbs were obviously lowered by medicated with 0.4, 0.2, 0.1 g/kg scorpion and centipede, as well as CII on the 32nd day after medication (P <0.05, P < 0.01). The expression levels of IL-2, IL-4, and IL-10 in the small intestine tissue homogenate were obviously lower in the model group than in the normal control group (P < 0.05, P < 0.01). Compared with the model group, only the expression levels of IL-2 and IL-4 in the small intestine tissue homogenate of the high dose scorpion and centipede group and the type II collagen treatment group significantly increased. The expression level of IL-10 significantly increased in the high and middle dose scorpion and centipede groups, as well as the type II collagen treatment group, showing statistical difference (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>Scorpion and centipede could effectively release the joint injury of rats with CIA. Its mechanism might be correlated with increased expression levels of IL-2, IL-4, and IL-10 in the small intestine mucosa.</p>