ABSTRACT
Objective:To study the protective effect of different doses of single-flavored Coptis， Magnoliae Officinalis Cortex， and their compatibility on ulcerative colitis （UC） model rats and the colonic B lymphoblastoma-2 associated X protein （Bax） and cysteine-containing aspartame-3（Caspase-3） protein， inflammatory cytokines， and other expressions. Method:The 120 healthy adult SD rats were randomly divided into blank group， model group， sulfasalazine group， Coptidis Rhizoma 2.00， 1.00， 0.50 g·kg-1 group， Magnoliae Officinalis Cortex 2.00， 1.00， 0.50 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00， 2.00， 1.00 g·kg-1 group， 12 groups with 10 rats in each group. The UC model was prepared by 2，4， 6-trinitrobenzene sulfonic acid/ethanol （TNBS/ethanol）. After 24 h of modeling， the rats were gavaged at 10 mL·kg-1 for one time/d. After modeling， the mental state， activity state， hair luster， stool characteristics， and blood in the stool of each group were observed. After continuous administration for 6 days， colon tissues and spleen were taken after the last administration for 24 h. The ratio of colonic weight to length and spleen index was calculated. The degree of colonic injury was evaluated according to the colonic mucosal injury index （CMDI） score criteria. the histopathological observation was performed using hematoxylin-eosin staining （HE）. The expression levels of tumor necrosis factor （TNF）-α， interleukin-6 （IL-6）， interleukin-10 （IL-10）， and myeloperoxidase （MPO） in the serum of Coptidis Rhizoma 2.00 g·kg-1 group， Magnoliae Officinalis Cortex 2.00 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 were detected by enzyme-linked immunosorbent assay（ELISA） in blank group and model group. Western blot was used to detect the expression of Bax and Caspase-3 proteins in the colon of rats. Result:Compared with blank group， rats in model group were sluggish and less active. The colon weight-length ratio， spleen index， CMDI， and colon tissue pathological damage increased significantly， and the expression of serum TNF-α， IL-6， and MPO increased significantly. Serum IL-10 expression levels were extremely significantly reduced （P<0.01）. Compared with model group， the sulfasalazine group， the Coptidis Rhizoma 2.00， 1.00 g·kg-1 group， the Magnoliae Officinalis Cortex 2.00 g·kg-1 group， and the three-dose groups of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex， their colon weight-length ratio and CMDI were significantly reduced （P<0.05，P<0.01）. The colon weight length ratio and CMDI index of the Coptidis Rhizoma 0.50 g·kg-1 group， Magnoliae Officinalis Cortex 0.50 and 1.00 g·kg-1 group were not significantly different from the model group but compared with Coptidis Rhizoma and Magnolia 0.50 g·kg-1 group， the ratio of colon weight to length in the group of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 1.00 g·kg-1 group was significantly reduced （P<0.01）. Compared with model group， the spleen index of the sulfasalazine group， the Coptidis Rhizoma 2.00 g·kg-1， and the Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 group were significantly lower （P<0.05）， compared with model group， the sulfasalazine group， Coptidis Rhizoma 2.00， 1.00 g·kg-1 and Magnoliae Officinalis Cortex 2.00 g·kg-1， thre dose groups of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex can significantly improve the depth and scope of histopathological damage and tissue necrosis. Compared with the model group， the preferred Coptidis Rhizoma 2.00 g·kg-1 group， Magnoliae Officinalis Cortex 2.00 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 group serum TNF-α， IL-6， MPO expression levels are extremely significantly reduced， the level of IL-10 increased significantly （P<0.01）．Compared with blank group， the expression of Bax and Caspase-3 protein in the colon of model group was significantly increased （P<0.01）. Compared with model group， the expression of Bax and Caspase-3 protein in preferred Coptidis Rhizoma 2.00 g·kg-1 group and Magnoliae Officinalis Cortex 2.00 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 group were significantly reduced （P<0.01）． Conclusion:The compatibility of single-flavored Coptidis Rhizoma， Magnoliae Officinalis Cortex， and Coptidis Rhizoma combine with Magnoliae Officinalis Cortex may improve the pathology of UC model rats induced by TNBS/ethanol by down-regulating the expression of Bax and Caspase-3 protein， inhibiting the release of inflammatory cytokines and promoting the release of anti-inflammatory factors injury， it plays a role in protecting colonic mucosa. The compatibility effect of Coptidis Rhizoma and Magnoliae Officinalis Cortex is better than that of single medicine， and Coptidis Rhizoma has a tendency to be better than Magnoliae Officinalis Cortex.
ABSTRACT
Aim To study the effect of the Kangfuxin liquid on 2,4,6-trinitrobenzene sulfonic acid (TNBS) induced ulcerative colitis in rats, and to explore its mechanism. Methods Male SD rats were divided into normal control, model control, SASP groups, and Kangfuxin low,medium and high dose groups. In ad-dition to the normal control group, other groups were induced ulcerative colitis with TNBS solution. Disease activity index (DAI), organ index, colon mucosa damage index (CMDI) and histopathological score (HS),the expression levels of IL-4, IL-17 in serum, and MPO, EGF, TGF-β1 in colonic mucosa were de-termined. Results The DAI score showed that the model was successful. Compared with the normal group,the level of IL-4 and IL-17, EGF and TGF-β1 in the model group were reduced significantly, while the CMDI score,HS score,colon index and MPO were elevated significantly. The DAI, CMDI, HS and MPO were reduced (P<0.05 or P<0.01), and the levels of IL-4, IL-17, EGF and TGF-β1 increased signifi-cantly(P <0.01). Conclusions Kangfuxin liquid can effectively alleviate ulcerative colitis induced by TNBS in rats. The mechanism may be related to the down-regulation of MPO expression and up-regulation of IL-4,IL-17,EGF and TGF-β1 levels.
ABSTRACT
Objective:To evaluate the therapeutic effects of sinomenine on T-helper cell type 1-mediated experimental colitis in-duced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in mice. Methods:Balb/c mice were divided into five groups:ethanol control group, TNBS model group, and sinomenine treatment groups (50, 100 and 200 mg·kg-1 ) with 10 ones in each. Colitis was induced by colonic instillation of TNBS dissolved in 0. 1 ml of 50% ethanol. Seven days after the colonic instillation of TNBS, sinomenine was given by gastric gavage once daily for 21 days. The mice were sacrificed on the 28th day, the injury degree of colonic mucosa was ob-served, the colon myeloperoxidase ( MPO) activity was determined, and the levels of inflammatory cytokines ( TNF-α, IL-17 and IL-23) were determined by ELISA. Results:Compared with those in TNBS model group, the body mass, gross injury score and histologi-cal findings in sinomenine groups at medium dose and high dose were significantly improved (P<0. 05), the activity of MPO signifi-cantly decreased (P<0. 05), and the protein levels of TNF-α, IL-17 and IL-23 in colonic mucosa were lower than those in TNBS group (P<0. 05). Conclusion:Sinomenine has notable therapeutic effect on TNBS-induced chronic colitis in mice, and the mecha-nism is related to the inhibition of Th1 cytokines by sinomenine.
ABSTRACT
<p><b>OBJECTIVE</b>To analyze the immunological characteristics of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis model and examine the therapeutic effects and mechanisms of Astragalus polysaccharides (APS) treatment.</p><p><b>METHODS</b>Thirty-two male specific pathogen free Spragne-Dawley rats were randomly equally assigned to four groups: control, TNBS, APS and prednisone groups. Experimental colitis was induced by enema administration of TNBS. Then rats were treated with APS (0.5 g•kg•day, once daily) or prednisone (1.0 mg•kg•day, once daily) by gavage for 14 days. Macroscopic lesion and histological damage were determined, and activity of myeloperoxidase (MPO) was measured in the colonic tissues. Expressions of T-box expressed in T-cells (T-bet) and GATA-binding protein-3 (GATA-3) were determined by immunohistochemistry analysis and western blot.</p><p><b>RESULTS</b>Both macroscopic lesion and histological colonic damage induced by TNBS were reduced by APS and prednisone treatment. These were accompanied by significant attenuation of MPO activity (P=0.03). TNBS intervention enhanced the expression of both GATA-3 and T-bet, but the expression of T-bet was significantly enhanced than that of GATA-3, resulting in significant reduction of GATA-3/T-bet ratio (P=0.025). APS administration enhanced the expression of T-bet (P=0.04) and GATA-3 (P=0.019) in comparison to TNBS group, and resulting in an up-regulated GATA-3/T-bet ratio. Prednisone treatment inhibited both expressions; however it also resulted in up-regulation of the GATA-3/T-bet ratio.</p><p><b>CONCLUSIONS</b>These results demonstrated that APS exerted a beneficial immune regulatory effect on experimental colitis. It promoted the expression of T helper cell 1 (Th1) and T helper cell 2 (Th2) specific transcription factors but ultimately favor a shift toward Th2 phenotype, suggesting that APS possessed therapeutic potential in experimental colitis.</p>