| With the development of global economy and the change of human lifestyle and diet structure,the incidence of hyperuricemia and the resulting gout(gouty arthritis)is increasing year by year,and the affected population is also getting younger,thus bringing a variety of concurrent systemic diseases and seriously affecting human health.The basic symptom of hyperuricemia and gout is the abnormal increase of blood uric acid,which is beyond the conventional fluctuation range and persists so high that urate crystals are deposited in joint synovium,bursa,cartilage and other tissues,leading to the attack and development of gout.At present,researches on gout mainly focus on purine metabolism.Although there are researches on immune regulation,they are still in the initial stage.Most of the works from the field of innate immunity reveal that the attack of gout is caused by urate microcrystals stimulating the activation of synovial endothelial cells,promoting the adhesion and exudation of mononuclear macrophages,and secreting inflammatory factors after phagocytosis of urate microcrystals.The process involves the increased content of various pro-inflammatory cytokines,etc.However,there are few studies on the imbalance of immune homeostasis in the earliest stage of gout(in the state of sustained high uric acid),the immune disorders in the stage of gout attack(deposition of urate crystal MSU into joints and other tissues),and the effect of high uric acid on the exacerbation of gout.At the same time,the medicinal and edible homologous materials of green plum flower were screened,and the advantages of enzymatic hydrolysis and probiotic fermentation transformation were integrated to obtain products with better activity,which could achieve the function of immune homeostasis repair while reducing uric acid,and the alleviating effect of green plum flower fermentation extract on gout and the possibility of its application in the treatment of gout in the future were discussed.The main research content of this thesis is as follows:(1)Gavage of 60 mg/m L xanthine +25 mg/m L ethambutol hydrochloride,injection of 20 mg/m L potassium oxazinate combined with 15 mg/m L xanthine +25 mg/m L ethambutol hydrochloride,The mouse model of high uric acid was established by three ways of gavage administration of 15 mg/m L xanthine +25 mg/m L ethambutol hydrochloride +20 mg/m L potassium oxazine,and the blood uric acid level was determined.The final choice was to gavage 60 mg/m L xanthine +25 mg/m L ethambutol hydrochloride.The histopathological analysis showed that the renal glomeruli of the mouse model showed no atrophy,the renal tubules were all dilated,and the renal interstitium showed no obvious inflammatory factor infiltration.The overall renal condition was basically no different from that of the control group,indicating that the model had no kidney damage,which met the requirements of using this model to study immune disorders caused by high blood uric acid.Flow cytometry analysis showed that the immune homeostasis of mice was broken under the condition of high blood uric acid,and the characteristic immune cells T cells(CD4 T cells and CD8 T cells),B cells,NK cells and myeloid cells of innate and acquired immunity showed varying degrees of changes compared with the control group,indicating that high uric acid can cause immune disorders in the body.By comparing the changes of characteristic immune cells in peripheral blood samples of people with high uric acid and those of normal control group,it was found that T cells(CD4 T cells and CD8 T cells),B cells,NK cells and myeloid cells in peripheral blood of people with high uric acid showed varying degrees of changes compared with those of normal people.The above results are very similar to those of the mice hyperuricemia model,confirming that high uric acid can cause the imbalance of immune homeostasis.(2)A mouse model of acute gout induced by sodium urate crystals was constructed,and the degree of pro-inflammatory and inflammatory infiltration of sodium urate crystals on mice at different times was screened.It was found that 48 h in the acute gout model could cause the differentiation and migration of the most inflammatory cells(CD45+).Therefore,48 h was selected as the observation point under the mouse model of acute gout.Compare the MSU-induced acute gout model of ordinary WT mice with the MSUinduced acute gout model of mice with high uric acid,and study the aggravation effect of high uric acid on acute gout.The experimental results show that the swelling degree of mice feet is more serious under the condition of high uric acid,and pathological results show that high uric acid aggravates the recruitment of inflammatory cells in the feet.Moreover,high uric acid further aggravated the immune disorders caused by MSU.CD45+ cells in the feet were significantly increased,and other characteristic immune cells also showed varying degrees of changes.The comparative study on the peripheral blood and draining lymph node data of mice under the two models also indicated the aggravation of high uric acid on acute gout.(3)The enzymatic hydrolysis extraction process of plum blossom was designed and optimized,and five strains(Lactobacillus Hirschi,Enterococcus faecium,acetobacterium,Aspergillus oryzae,Mucor saurus)were used to ferment enzymatic hydrolysis extracts of plum blossom,and it was found that the extracts of plum blossom fermented by Lactobacillus Hirschi had better anti-inflammatory and uric acid lowering effects.(7)The extracts of plum blossom fermented by Lactobacillus Hirschi screened in the experiment were used.To solve the problem of immune disorders caused by high uric acid and exacerbating gout,the experimental results showed that in addition to effectively reducing uric acid,the extracts of plum blossom fermented by Lactobacillus Hirschi could also repair the homeostasis of T cells(CD4 T cells and CD8 T cells),B cells,NK cells and myeloid cells in mice with high uric acid to varying degrees(compared with the control group).In the acute gout model,Lactobacillus Hirschi fermented plum flower extract could relieve the foot swelling of mice in the acute gout model,but could not restore the immune homeostasis of the foot,which was speculated to be related to the metabolic process of plum flower extract and the chemotactic effectIn summary,in this thesis,by optimizing the enzymatic hydrolysis extraction process of plum blossom,the extract of plum blossom was produced by bacterial fermentation,and a mouse model of hyperuricemia and acute gout was established.The imbalance of immune homeostasis in hyperuricemia and the exacerbation of immune disorder in acute gout were identified,and the important role of immune homeostasis in the occurrence and development of hyperuricemia/gout was studied.It was found that the extracts of plum blossom fermented by Lactobacillus Hirschi had good anti-inflammatory and uric acid lowering functions,and could relieve the immune disorders caused by high uric acid,which would be helpful for the development of medicine and food homology products for the treatment and relief of gout in the future. |
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