Establishment Of A Mouse Model Of Gout And The Study Of Physiological Function

Backgroundand Objective:Gout is a heterogeneous metabolic disease with increased blood uric acid caused by long-term purine metabolism and uric acid excretion disorders.When the physiological saturation threshold for uric acid is exceeded in body fluids,urate crystals will be formed and deposited in the joints and other tissues.The clinical manifestations of MSU crystal deposition include acute and chronic arthritis,joint deformity,tophi and so on.MSU crystal is also often implicated in chronic interstitial nephritis and uric acid stones,which can lead to acute and chronic kidney failure.Gout is often associated with obesity,hyperlipidemia,hypertension,type 2 diabetes,cardiovascular diseases and other diseases.Genetic factors,sex hormones,diet and alcoholcan promote the occurrence and development of gout.Gout has seriously affected quality and level of our life.In recent years,the prevalence of gout is rising year by year,and it has become a threat to human health,and the occurrence of gout has geography,age and gender differences.Current animal models for study of gout include acute gouty arthritis animal model,gouty nodules and swelling animal model and hyperuricemia model.Injecting sodium urate intoarticular cavity of animal models directly is the commonest way for gouty arthritis.But this model is different from human pathogenesis of gout.There is no hyperuricemia animal model accompanied by gouty arthritis.The research of exploring and building this animal model will get great value.Experimental results show that testosterone may increase renal reabsorption of uric acid,but the mechanism of testosterone involved in the pathogenesis of gout is unclear.Although there are some clinical drugs which can effectively control the symptoms of gout,serious side effects limit their use.The research of testosterone on the pathogenesis of gout can provide new targets for gout treatment.1.Establishing mice model of hyperuricemiaAnimal models were given 300 mg/kg oxonic acid potassium salt through intraperitoneal injection.It inhibitedthe activity of uric acid enzyme causing elevated levels of serum uric acid.Collected the blood of mice during different time periods,and measuredthe values of uric acid.The results showed that the mice hyperuricemia model has been successfully established.Serum uric acid levels increased 1 hour later after injecting oxonic acid potassium salt,reached the highest levels at 4 hours after injection,and began to decline at 6 hours after injection.2.Establishing mice model of goutAnimal models were given 300 mg/kg oxonic acid potassium salt through intraperitoneal injection everyday.Every other five days injected 20 μL 1%acetic acid into the mice hind limb joints,and observed hind limb swelling during this time.The hind limbs of mice were used to make paraffin pathological section.After methenamine silver staining we observed if there was deposition of sodium urate crystal in the joints.The results showed that after continuous injection oxonic acid potassium salt for 3 months,individual mice had obvious hind limb joint swelling.Similar sodium urate crystalline material was taken from hind limb parts,and it was found that it contained sodium urate with dissolved test substance.Paraffin sections of hind legs were performed,then methenamine silver staining was applied.Paraffin pathological section of mice model kidney was also made to investigate the effects of long-time hyperuricemia on kidney pathological changes.The results showed that long-term injection oteracil potassium can cause mouse glomerular wall thickening,interstitial inflammatory cell infiltration and tubular epithelial cell swelling.3.Measuring the degree of paw swelling in mice and test the thermal sensitivity and graspingability.Experiments showed that,comparing with the control group mice,mice model have shown that thickness of the foot increased,thermal sensitivity raised and grasping ability reduced.4.Studying the relationship between hyperuricemia and testosteroneIn this experiment,we wanted to investigate the effect of sodium urate on testosterone secretion capacity by stimulating mouse primary testicular interstitial cell with sodium urate.After cells incubated with sodium urate for 24 hours on the condition of 37℃ and 5%CO2,then the culture medium was tested for testosterone levels.The results showed that mouse testicular interstitial cells stimulated by different concentrations of sodium urate did not affect the secretion of testosterone.Therefore sodium urate did not affect the secretion of testosterone by mouse testicular interstitial cell.5.Exploring the influence of hyperuricemia on luteinizing hormoneIn this experiment,we choosed mice with continuous injection of oxygen hydrogen potassium solution for 30 days.And investigated the influence of hyperuricemia on luteinizing hormone level.LH ELISA kit was used to detect the content of LH in serum.The results showed that the hyperuricemia mouse model has been established.Based on this model,the gout disease has been induced successfully by long-term continuous intraperitoneal injection of the urate oxidase inhibitors oteracil potassium,which not only kept uric acid at a high level for a long time,but also destroyed the normal pH environment in mouse arthrosis,causing the free sodium urate crystallization deposited in the joints more easily.The article showed that mouse gout models have been successfully established by observing the behaviors and limbs pathological slices of mice,and found LH plays an important role in causing male mice suffering from gout...