Bioinformatics Based Gene Analysis Of Chronic Renal Injury Induced By Hyperuricemia

Hyperuricemia(Hyperuricemia,HUA)is a chronic metabolic disease caused by purine metabolism disorder.HUA was considered to have serum uric acid(Serum Uric Acid,SUA)concentrations>7 mg/d L(>420 μM)in men,SUA concentrations> 6mg/d L(>360 μM)in women,and child and adolescent SUA concentrations of 5.5mg/d L(>330μM).An epidemiological survey showed that the prevalence of HUA was higher than 20.1% in America and higher than 17.7% in China.The prevalence of HUA has increased in recent years.HUA can cause and induce a variety of diseases,including gout,cardiovascular disease,and chronic kidney injury(Chronic Kidney Disease,CKD).The kidney is the main excretion site of uric acid(Uric acid,UA),and about 90% of HUA patients are caused by the renal excretion dysfunction of UA.However,HUA is not easy to be detected in the early stage,and continuous progression can cause irreversible damage of renal function.Therefore,the CKD caused by HUA is widely concerned.Studies have shown that the pathogenesis of CKD caused by HUA mainly includes the crystal formation of single sodium urate salt(Monosodium Urate,MSU),oxidative stress,renal fibrosis and inflammation.Currently,the mechanism of HUA causing CKD is complex and controversial,and the underlying molecular mechanisms are also worth exploring.In conclusion,this project intends to preliminarily explore the mechanism of CKD induced by HUA.Objective:Observe the dynamic changes of kidney injury during HUA,use bioinformatics technology to analyze and predict the key molecules,and further verify the key molecules,laying a foundation for the mechanism of HUA induced CKD.Methods:1.The mouse model of hyperuricemia was prepared by intraperitoneal injection of potassium oxyazate and hypoxanthine.2.The experiment was divided into Control and Model groups(administered 3W,5 W,7 W,9 W,11 W,13 W and 19 W).3.SUA levels were detected after 1 h of administration using a phosphotungsten serum uric acid kit.4.HE staining.5.Mice was used to detect renal fibrosis by Masson staining.6.Use a polarized light microscope to detect the MSU crystal deposition index.7.Test HUA and CKD related Hub genes by bioinformatics,and use CKD related genes on databases GSE190205 and Dis Ge NET to conduct differential expression analysis,screen differentially expressed genes(differentially expressed genes,DEGs),conduct KEGG,GO enrichment analysis and PPI network.8.Hub gene and protein expression were detected by PCR and Western blot,respectively.Results:1.The Model group SUA was three times higher than the Control group.2.HE staining found that kidney damage began in the Model group at the 5th week,which was aggravated with the prolonged course of the disease.The main manifestations of injury are atrophy of renal tubules,inflammatory cell infiltration,dilated renal tubules,glomerular atrophy and loss of function.3.Masson The staining showed mild fibrosis in the kidneys in the fifth week,which worsened with the duration of the disease.4.Polarized light microscopy revealed MSU crystals since week 9.5.The functions enriched by credit analysis are mainly the regulation of inflammation,including Hub genes including Il-6,Il-1β,Ccl2,Ccl5,Cxcl10,Itgam,Stat3 and Tnf-α.6.PCR results showed that the expression of Ccl5,Ccl2,Cxcl10 and Tnf-α gene increased since week 5;Il-6 gene increased since week 9;Stat3 gene increased since week 7 weeks;and Il-1β gene increased since 11 weeks,and the expression level gradually increased with the duration of the disease.7.Western blot results showed that the expression of CCL 2 and CCL 5 protein increased since week 5;the expression level of CXCL10 protein increased since week7 th,and the expression level gradually increased with the duration of the disease.Conclusion:1.With prolonged HUA in hyperuricemia in mouse model,high levels of blood uric acid through the inflammatory response,fibrosis and MSU crystals cause altered morphogenesis of renal tubules and glomeruli leading to the occurrence and development of chronic kidney injury.2.proved that the increased expression of Il-6,Il-1β,Ccl 2,Ccl 5,Cxcl10,Stat 3and Tnf-α m RNA was observed in the mouse hyperuricemia model,which may participate in the regulation of HUA-induced kidney disease in mice via inflammatory response.3.The Ccl2,Ccl5,and Cxcl10 genes play a key role in hyperuricemia-induced chronic kidney disease.