Study On The Mechanism Of Diosmetin In Preventing And Treating Uric Acid Nephropathy

Hyperuricemia nephropathy(HN),also known as gouty nephropathy and hyperuricemia nephropathy,is a renal injury disease caused by abnormal purine metabolism.Hyperuricemia(HUA)is one of the most common metabolic diseases in the world,caused by increased uric acid production or decreased excretion.Developing effective strategies to prevent the progression of uric acid nephropathy and improve renal function is crucial to reducing the burden of disease.Diosmetin is a flavonoid compound with anti-inflammatory,antioxidant,and anti-tumor activities.The previous research results of our research group indicated that diosmetin had a preventive and therapeutic effect on uric acid nephropathy in mice,but there was no in-depth study on its mechanism.In this study,a model of renal tubular epithelial cells(HK-2 cells)injury induced by monosodium urate(MSU)was established in vitro,and a model of uric acid nephropathy induced by yeast extract combined with potassium oxyzinate was established in mice in vivo to explore the mechanism of action of diosmetin.(1)In vitro studies,HK-2 cells were pretreated with 5,10 and 20 μM of diosmetin for 2 h and 2 mg/m L MSU was added to induce cell damage.The results showed that diosmetin significantly reduced the levels of nitric oxide(NO)and lactate dehydrogenase(LDH)in the cell supernatant.The results of protein blot experiments and RT-q PCR showed that diosmetin significantly inhibited TLR4,NLRP3,ASC,Caspase-1,IL-1β,IL-18,PI3 K,AKT,NF-κB protein and gene expression reduced the inflammatory damage induced by MSU,and by enhancing the ratio of Bcl-2 / Bax in HK-2 cells,and inhibited Caspase-3 protein and gene expression reduced HK-2 cell apoptosis.(2)In vivo research,a mouse model of uric acid nephropathy was established using 30 g/m L yeast extract combined with 300 mg/m L potassium oxazinate.The mechanism of diosmetin against uric acid nephropathy was studied through intragastric administration.The results showed that diosmetin(20 mg/kg,40 mg/kg,80 mg/kg)reduced the levels of serum uric acid,creatinine,and urea nitrogen,improved renal inflammatory reaction,renal tubular necrosis and expansion,and renal fibrosis in mice with uric acid nephropathy.The results of protein blot experiment and q PCR showed that diosmetin inhibited the expression of TLR4/NLRP3 and PI3K/AKT/NF-κB pathway protein and mRNA,and reduced inflammation and apoptosis;diosmetin also regulated ABCG2 and GLUT9 protein and genes expression in kidney and intestinal tissues of uric acid nephropathy.The species richness of the HN mouse gut measured by 16 s RNA showed that diosmetin could modulate species richness in mouse feces.In summary,diosmetin can reduce serum uric acid,creatinine,and urea nitrogen levels.Different concentrations of diosmetin significantly decrease the levels of NO and LDH secreted by HK-2 cells,inhibite inflammation and apoptosis by adjusting TLR4/NLRP3 and PI3K/AKT/NF-κB pathways,upregulate ABCG2 and downregulate GLUT9 uric acid transporters,and improve intestinal species richness to treat uric acid nephropathy.