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Objective To observe the protective effect of probiotics VSL # 3 on the rats of trinitrobenzene sulphonic acid (TNBS) induced acute colitis, and the effect on the expression of tight junction proteins claudins (claudinl, 2, and 3) in colon mucosa. Methods Rat of colitis model was induced by TNBS one time enema. Since the second day after the model established, 2ml 0.9％ sodium chloride was intragastric administrated into 8 rats each day (TNBS group), 8 rats with 100 mg VSL # 3 (VSL # 3 group ), another 8 rats with 5-aminosalicylic acid (5-ASA) intragastric administration (5-ASA group), and 0.9％ sodium chloride was intragastric administrated daily into 5 no modeling rats as control group. All were treated for 7 days and then sacrificed. Fecal samples were collected for bacterial culturing and detecting the changes of colon bacteria colonies. Disease activity index (DAI was calculated according to diarrhea, hemafecia and weight. The colon tissue was taken for HE staining and the pathological changes were observed under microscope.The changes of myeloperoxidase (MPO) level in colon tissue were examined by enzyme-linked immunosorbent assay (ELISA). The expression and distribution of claudins 1, 2, and 3 in colon tissue was detected by a novel quantum dots immunofluorescence labeling technique. Results Compared with TNBS group,both VSL # 3 and 5-ASA treatment could improve colitis, as well as decreas DAI and colonic MPO level, and lower the inflammatory score of colitis. Compared with control group, the bacteria counting in the colon of colitis rats changed, the expression of Claudins 1, 2, and 3 expression was significantly increased (average density: 66.200±5.737, 71.780±6.670 and 61.300±5. 199,t＝17.237, 27.909and 21.788, all P＜0.01). Compared with TNBS group or 5-ASA group, VSL# 3 could adjust the balance of intestinal bacteria and increase the expression of Claudin 1 and 3 (Claudin 1 : 75. 550 ±8.717, 66.200±5.737 and 67.080±5.401;t＝9.348, 8. 469; all P＜0.05; Claudin-3: 68. 820±7.443, 61.300±5. 199 and 59. 830±5. 930;t＝7. 519, 8. 988, all P＜0.05), however claudin 2 expression was down-regulated (58. 740±6. 457, 71. 780±6. 670 and 66. 870±5. 791; t＝ 13. 033,P＜0.01, t＝8. 123, P＜0. 05). Conclusion Probiotics VSL# 3 may enhance the intestinal mucosal barrier function through changing the claudins expression, and play a role in the relief of inflammatory reaction in acute colitis.
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BACKGROUND/AIMS: Trinitrobenzene sulphonic acid (TNBS)-induced colonic inflammation in rat alters colonic motor function. The aims of this study were to investigate the effect of TNBS-induced colitis on the colonic motor function of a guinea pig along the course of colitis and to document persistently altered colonic motor function after resolution of inflammation. METHODS: Colitis was induced in about 300 g male guinea pigs (Hartley) by intrarectal administration of 0.3 mL TNBS in 50% ethanol, while controls received 0.3 mL of 50% ethanol or not. After 24 hours, 48 hours, 72 hours, 1 week, and 2 weeks, the distal colon was taken for the investigation of gross and microscopic findings, muscle tension, and colonic transit. RESULTS: Maximal mucosal injury and inflammation were evident from the 2nd day following the induction of colitis. Seven days after the induction of colitis, some portions of the damaged mucosa began to recover. Development of tension in response to carbachol was not altered significantly along the course of colitis. Colonic transit was delayed significantly at 3, 7, and 14 days after administration of TNBS. CONCLUSIONS: Colonic transit in TNBS-induced colitis of guinea pigs is delayed, but not linearly related to the degree of inflammation. Delayed colonic transit is not related to the muscarinic receptor-mediated contractions of circular muscle from the inflamed colon. Further studies are required to determine the mechanism(s) involved in this motor change.
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Animais , Humanos , Masculino , Ratos , Carbacol , Colite , Colo , Etanol , Cobaias , Guiné , Inflamação , Mucosa , Tono MuscularRESUMO
Objective To investigate the therapeutic effects of curcumin(Cur)on trinitrobenzene sulphonic acid(TNBS)induced colitis and to investigate cytokines change in colon mucosa,spleen and sera.Methods Colitis was induced in SD rats by intrarectal injection of TNBS(100 mg/kg).Experi- mental animals were divided into negative control group,TNBS group and Cur therapeutic group(Cur, 30 mg?kg~(-1)?d~(-1),intraperitoneal injection).Expression of cytokines mRNA in colon mucosa was observed by RT-PCR,intracellular cytokines of interferon(IFN)-?and interleukin(IL)-4 in splenocytes were detected by flow cytometry(FCM),concentrations of IFN-?and IL-4 in sera were determined by enzyme-link immunosorbent analysis(ELISA).Results After treatment with Cur,macroscopic scores (3.9?1.0 vs.2.2?0.7),myeloperoxidase(MPO)activity(15.0?2.6 vs.7.3?1.4),mRNA of IFN-?(1.02?0.07 vs.0.06?0.02,mRNAof IL-12(0.29?0.05 vs.0.11?0.01)and the ratio of IFN-?/IL-4(11.44?0.97 vs.0.38?0.10)in colon mucosa,proportion of IFN-?CD4~+(31.7?7.5 vs. 21.1?3.7)and the ratio of IFN-?/IL-4 CD4~+(19.9?5.1 vs.6.1?1.8)in splenocytes,concentra tions of IFN-?[(1528?159)pg/ml vs.(513?14)pg/ml] and the ratio of IFN-?/IL-4(19.5?4.1 vs. 4.2?0.6)in sera were all decreased(P＜0.05 or P＜0.01).Meanwhile,mRNA of IL-4(0.09?0.01 vs.0.15?0.04)and IL-10(0.28?0.08 vs.0.63?0.12)in colon mucosa,proportion of IL-4 CD4~+ (1.6?0.5 vs.3.4?1.1)in splenocytes and concentrations of IL-4 in sera[(81?15)pg/ml vs.(124?20) pg/ml] were all increased after the treatment of Cur(P＜0.05 or P＜0.01).Conclusions Cur showed ther- apeutic effects on TNBS-induced colitis,and the mechanism might be through regulating the balance of Th1/ Th2 in colon locally and systematically.
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Objective:To study the expression of P selectin in the colonic mucosa and on surfaces of platelets in circulation, and determine its association with IL 8 and TNF ? in trinitrobenzene sulphonic acid (TNBS) induced colitis in rat, in order to investigate the effects of P selectin on inflammation and thrombosis in inflammatory bowel disease Methods:TNBS was instilled into the colon of the rats to induce colitis Animals were sacrificed at the 24th hour, the 1st and the 2nd week after production Blood and colonic tissues were obtained P selectin on the platelets were detected by radioimmunoassay IL 8 in serum was detected by ELISA method Expression of P selectin and TNF ? in colonic mucosa were assessed with immunohistochemistry Results:P selectin in mucosa was increased in different time points compared with control group The positive expression was above 50% P selectin on the platelets wasn't increased at the 24th hour, but increased at the 1st and 2nd week (P