| Celiac disease is an inflammatory bowel disease caused by an autoimmune response.In individuals with genetic susceptibility genes,gluten protein peptides stimulate T cells of the immune system,which in turn triggers a complex immune response involving the innate immune system and the adaptive immune system.Currently,the best way to treat this autoimmune disease is for patients to adhere to a gluten-free diet throughout their lives.In food production and processing,wheat and its gluten protein have been used as raw materials or ingredients for a long time and extensively,resulting in patients inevitably passively ingesting gluten protein in their daily diet.Therefore,it is of great significance to explore new prevention and nutrition intervention methods to apply to such diseases.Intestinal inflammation and immune response in diseases such as celiac disease is related to oxidative stress,and antioxidants can effectively inhibit the oxidative response of free radicals,thereby alleviating oxidative stress and slowing down the inflammatory response.Whether the intake of natural antioxidants can be used as a new dietary intervention therapy to alleviate the related symptoms of celiac disease is the main purpose of this research.The main research content of this thesis includes the following four parts:(1)Construction of the intestinal inflammation model of C57BL/6 mice induced by wheat gluten protein;(2)Interventional effect of natural antioxidants on mice intestinal inflammation;(3)Molecular mechanism of resveratrol intervention on mice intestinal inflammation;(4)The design of resveratrol soft capsule factory with an annual output of 5 tons.The main research methods,results and conclusions are as follows.1.The C57BL/6 mice were fed with a gluten-free diet and raised to the second or more generations.4 to 6-week-old female mice were selected and gavaged with non-gluten ATI and gluten Gliadin,and further immunostimulated with CT,Poly:IC adjuvant and DSS,respectively,to construct a mouse model of intestinal inflammation.The results showed that the combined effect of ATI and Gliadin caused villi atrophy and crypt hyperplasia in the intestine of mice and increased the Th1/Th2 ratio in MLN and TG2 content in jejunal tissue.However,the symptoms were further aggravated after the addition of Poly: IC.The co-stimulation of ATI,Poly:IC and Gliadin caused a significant reduction in body weight,severe villous atrophy and crypt hyperplasia in the intestine,a shift in the Th cell population of MLN towards Th1-type immune cells and overexpression of relevant inflammatory factors in the jejunal tissue(IL-1β,IL-2,TNF-α,IL-4,IL-6,IL-10),and these symptoms and indicators in mice are closest to those of celiac disease.The mouse intestinal inflammation model constructed by co-stimulation of ATI,Poly:IC and Gliadin can be used as the experimental animal model for celiac disease research.2.Using the established model of intestinal inflammation in mice,four natural antioxidants selected for resveratrol,apigenin,curcumin and quercetin were treated with nutritional intervention in mice.The best natural antioxidants for nutritional intervention were screened using indicators of immunomodulation and oxidative stress.The results showed that resveratrol,apigenin,curcumin and quercetin could alleviate intestinal inflammation in mice,significantly improve the apparent symptoms and intestinal injury,regulate the distribution of Th cell subpopulations and the expression of related inflammatory factors(IL-1β,TNF-α)and reduce the content of TG2 in jejunal tissue.Among them,resveratrol and quercetin were the most effective in immunomodulation of intestinal inflammation in mice,which play a significant role in improving intestinal injury and regulating Th cell subpopulation toward Th2 differentiation,inhibiting Th1 immune cell function overexpression(IL-1β,TNF-α),slowing Th2 immune cell function underexpression(IL-4,IL-6,IL-10)and reducing TG2 content in jejunal tissues.Furthermore,in the regulation of oxidative stress(NO,GSH,MDA content and SOD activity)in jejunal tissues of mice with intestinal inflammation by resveratrol,apigenin,curcumin and quercetin,resveratrol showed the strongest antioxidant efficacy,followed by curcumin.In conclusion,the nutritional intervention effect of resveratrol is best.3.Total RNA extracted from mouse jejunal tissue and transcriptome sequencing technology(RNA-seq)was used to analyze the differences in gene expression after resveratrol nutritional intervention in order to explore the regulatory mechanisms of resveratrol in celiac disease intestinal intervention.The study found that among the differential genes of resveratrol nutritional intervention in mice with intestinal inflammation,six significantly down-regulated genes(Orm1,Nr0b2,Fbxo27,Fgf15,Fabp4,and Amy1)and three significantly up-regulated genes(Ubd,Lat,and Aire)were associated with inflammatory,immunomodulatory,and oxidative stress responses.In the differential gene enrichment analysis,the differential genes were mainly related to the reaction of oxygenated compounds,phylogeny and reaction with organic nitrogen compounds in biological processes.In the broad category of biological systems,they were mainly associated with the enrichment of PPAR signaling pathway.In the broad category of metabolism,they were mainly enriched in nitrogen metabolism and in the broad category of environmental information processing,they were mainly displayed in the enrichment of AMPK signaling pathway and Fox O signaling pathway.4.In accordance with the relevant requirements of health food production,the processing plant design of resveratrol soft capsule was carried out with grape skin/seed as the main raw material.The production process route,main equipment selection and asset allocation of the resveratrol soft capsule plant were determined,while the plant floor plan and workshop floor plan were drawn,and the design of the processing plant with an annual capacity of 5 tons of resveratrol soft capsules was finally completed. |
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