| Objective:To study the therapeutic effect and mechanism of curcumin chitosan microspheres on ulcerative colitis.Methods:120 male SPF BALB/c rats were randomly divided into normal control group,model group,positive control group,curcumin group,chitosan microsphere group,and curcumin chitosan microsphere group,with 20 rats in each group.No treatment was given to the normal control group,and 5% dextran sodium sulfate(DSS)for 7d was used to induce UC model in other groups.Normal control group normal drinking water and diet,model group: 5%DSS free drinking water,normal diet;Positive control group: 5%DSS free drinking water + feed containing 5g/L salazopyridine;Chitosan microsphere group: 5%DSS free drinking water + feed containing 20g/L chitosan microspheres;Curcumin group: 5%DSS free water + feed containing 20g/L curcumin;Curcumin chitosan microspheres group: 5%DSS free water + feed containing 20g/L curcumin chitosan microspheres.Enzyme-linked immunosorbent assay to detect serum interleukin 1β,interleukin 6 and interleukin 4 levels,detection of chemical dyeing immune group NF-κB P65 colonic mucosa and the expression of ICAM-1,were observed under electron microscope and light microscope histopathological and ultrastructural changes of rat colon damage fraction of colonic histology,assessment of disease activity index score.Results:Ⅰ.enzyme-linked immunosorbent assay was used to determine the contents of IL-1β and IL-6 in the serum of the model group(71.13 2.58pg/ml),(77.67 6.48pg/ml),and IL-4(49.12 8.46pg/ml),while the contents of IL-1β,IL-6,and IL-4 in the positive control group were(52.11 1.43pg/ml),(55.37 5.31pg/ml),(60.11 6.220pg/ml),respectively.The contents of IL-1β,IL-6,and IL-4 in the serum of the curcumin groupwere(41.44 1.89pg/ml),(36.79 7.98pg/ml),(73.71 5.99pg/ml),and(59.79 1.78pg/ml),(56.71 5.31pg/ml),(65.39 9.63pg/ml),respectively.The contents of IL-1β,IL-6,and IL-4 in the serum of the rats in the curcumin chitosan microsphere group were(27.12.59pg/ml),(25.83 9.15pg/ml),(89.98 8.14pg/ml),and(24.33 2.87pg/ml),(30.37.69pg/ml),and(94.43 8.52pg/ml),respectively.There was no significant difference in IL-4 content between the curcumin chitosan microsphere group and the normal group(P > 0.05),which was significantly lower than that of other control groups(P <0.05).Ⅱ.Observation under light microscope: no inflammatory exudation such as lymphocytes and plasma cells was observed in the normal group,no ulcer surface was observed,the glandular structure was clear,and the mucosa was orderly,complete and smooth.In the colonic tissues of the model group,partial mucosal necrosis was observed,forming ulcers of varying degrees of depth.There were a large number of acute and chronic inflammatory cells in the lamina propria,intrinsic glandular crypt abscess formation,some glandular hyperplasia formed polypoid structures,and lymphocyte aggregation in the basement.The above phenomenon was reduced in the positive control group and chitosan microsphere group,and was less in the curcumin group and least in the curcumin chitosan microsphere group.Ⅲ.Were observed under electron microscope: model group the mucosal surface and crypt epithelial cells can be destroyed,a significant increase in the gland fossae goblet cells significantly emptying,mucosal crypt in increase in the number,some fossae shape distortion,part fossae irregular surface,obvious expansion of endoplasmic reticulum,mitochondria markedly swollen,reduced numbers of cell membrane aperture,a large number of lymphocytes,can be seen in the stroma glands goblet cells decreased,focal crypt abscess formation.The above phenomenon was not observed in normal rats,but in the positive control group and chitosan microsphere group,the above phenomenon was reduced;in the curcumin group,the above phenomenon was mild;in the curcumin chitosan microsphere group,the above phenomenon was not obvious.Ⅳ.Immunohistochemical assay: NF-κB P65 proteins activated in rat intestinal tissues are transferred to the nucleus,while those not activated remain in thecytoplasm.So after HE staining,can be observed in the nucleus and cytoplasm can be shaded into tan,the NF-κ B P65 protein expression more colonic mucosa lamina propria of large amounts of mononuclear cell nucleus color is deeper,the NF-κB P65 less protein expression of colonic mucosa lamina propria of large amounts of mononuclear darker cytoplasm,occasionally also saw a few colon,tan epithelial cell nucleus or cytoplasm.However,positive ICAM-1 expression was mainly concentrated in the cytoplasm of leukocytes,neutrophils and proliferative vascular endothelial cells in the colonic mucosa.The cytoplasm of these stained inflammatory cells was tan.These proliferating blood vessels are mainly concentrated in the colonic mucosa,and occasionally a small amount of low ICAM-1 expression is found in a small number of colonic epithelial cells.The positive expression of NF-κ B P65 and ICAM-1 was not obvious in the normal control group(10.52 3.51%,9.65 1.36%).The nuclear staining of NF-κB P65 protein was the most obvious in the model group(52.12 8.81%).The chitosan microsphere group was(44.11 5.95%),(53.40 8.79%),the curcumin group was(13.02 4.52%),(15.30 3.95%),and the curcumin chitosan microsphere group was(11.10 3.68%),(10.05 2.35%).It can be seen that the positive expression rate of curcumin chitosan microsphere group was not statistically different from that of the normal control group(P > 0.05),which was significantly lower than that of other groups(P < 0.05).Ⅴ.Disease activity index score: after successful modeling,the activity of the rats was decreased,and the rats were lazy,and the amount of diet was decreased,and the body weight was significantly and continuously reduced,and blood could be seen in the stools with naked eyes.From the intervention to the end of treatment,the disease activity index score of the model group(7.03 1.09),the positive control group(4.221.02),the chitosan microsphere group(6.12 1.11),the curcumin group(2.43 1.09),the curcumin chitosan microsphere group(0.46 1.16),and the normal control group(0.21 0.45).The disease activity index score of curcumin chitosan microsphere group was not statistically different from that of the normal control group(P > 0.05),which was significantly lower than that of other groups(P < 0.05).Conclusions:1.Curcumin chitosan microspheres have more significant effect in the treatment of ulcerative colitis than curcumin and chitosan microspheres alone,and have better treatment effect than salicylic acid drugs.2.it may reduce the inflammatory response by inhibiting the expression of NF-κB P65 and reduce the damage of colon tissue in mice with ulcerative colitis by anti-inflammatory effect. |
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