| Objective:Study was designed to determinate the compositions of the adlay seedextract(ASE), and to research the protective effects of ASE on ulcerative colitis ofWistar rats induced by dextran sulfate sodium, as well as to discuss the possiblemechanism from antioxidant and immune regulating.Methods：1. Determination of the ASE: Used Kjeldahl method, Liquid chromatographymethod, Soxhlet extraction method, gas chromatography assay to detect thecontent of protein, amino acids, fat and fatty acid respectively. Took enzymaticgravimetric method to assay the dietary fiber, atomic absorption spectrometry todeterminate ferrum and zinc, EDTA method and fluorescence method to set outcalcuim and selenium. Gravimetric means was used to determinate ash.2. Study of ASE effects on UC: Rats in model(DSS) group and the ASEtreatment group were fed with5%DSS solution to induce ulcerative colitis. Therats in ASE group received treatments by gastric lavage daily with ASE, while theother two groups with distilled water from the beginning. DAI score, Macroscopicand histological assessment, IWT and immune organ coefficient were used toevaluate the general situation and the severity of colonic damage. The activity ofserum MPO, as well as SOD, GSH-PX, T-AOC, and the content of MDA in thecolon tissues and serum were measured by biochemical methods. ELISA waschosen to detect the serum TNF-α levels. The expression of HSP70and HSP70mRNA in the colon were reflected by means of immune histochemical and realtime quantitative RT-PCR.Results:1. ASE composition: Protein,12.4%; Sum of the amino acids,11.62%(17kinds of amino acids, mainly glutamic acid, alanine, leucine); Fat,8.15%;6kindsof clear fatty acid (mainly oleic acid, linoleic acid, palmitic acid); Total dietaryfiber,15%(soluble dietary fiber,0.58%); Ferrum,2.78mg/100g; Zinc,1.16mg/100g; Calcium,99.30mg/100g; Selenium,4.07μg/100g; Ash,0.645 mg/100g.2. DAI score of DSS group was higher than the other two groups. The ASEgroup’s DAI score was in a middle level of the three groups, and decreasedgradually after ASE treated. The damage of colon was more serious in DSS group,and IWT tended thicker. Colon damage could be mitigated after ASE treated. MPOactivity sharply decreased in ASE group compared with DSS group(P<0.05).3. No matter in tissue or serum, SOD and GSH-PX activity in ASE groupwere significantly higher than DSS group, and serum MDA content was less.4. Serum TNF-α expression levels in DSS group was higher than that ofnormal group (P <0.05), while TNF-α in ASE group was lower than DSS group (P<0.01) but higher than it of normal group (P <0.01).5. IOD of HSP70in DSS group was higher than that of the normal group (P<0.05). And IOD of HSP70in ASE group was significantly higher than the othertwo groups (P <0.01).6. The result of HSP70mRNA expression level of three groups was consistentwith HSP70expression level tested by means of immune histochemical that wasthe level of ASE was highest and DSS showed a middle level.Conclusion:ASE can significantly improve the UC general status and inflammatory injury.Meanwhile it can reduce the content of MDA and enhance the activity of SOD andGSP-PX. ASE shows the capacity of up regulating the expression level of HSP70and inhibiting TNF-α expression in addition. In summary, ASE can effect on UC ofWistar rats induced by DSS as a protection via anti-oxidative role and immuneregulating role. |
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