Changes Of Zinc Alpha 2 Glycoprotein In Renal Tissue Of Mice Model And HK-2 Cells Based On Uric Acid

Objective:In the mouse model of hyperuricemia based on high uric acid environment in mouse kidney zinc alpha 2（zinc alpha 2 glycoprotein glycoprotein,ZAG）changes in uric acid by constructing cell model,observe the change of ZAG protein in HK-2 cells of uric acid environment,to provide new ideas for the study of metabolic acidosis with high urinary diseases the.Methods:1)60 SPF Kunming mice were randomly divided into model group and control group,97%potassium oxonate by 0.5g/（kg/D）intraperitoneal injection to establish hyperuricemia mice model of uricase inhibition,7d modeling and 14d,detection of serum uric acid and blood from caudal vein（SUA）to determine the level of model successful establishment and detection of serum biochemical indexes in mice,the expression of Western Blotting and real-time fluorescence quantitative PCR detection of ZAGmRNA and ZAG.2)1640 containing 10%fetal bovine serum culture medium,culture box at 37 C（5%CO₂）in the recovery and passage of HK-2 cells,the experiment is divided into the following sections:（1）on human renal tubular epithelial cells stimulated by zinc at the same concentration of uric acid 2-glycoprotein（ZAG the expression of dynamic）:the experiment was divided into blank control group,hyperuricemia group,namely:0.1mg/ml,0.21mg/ml and 0.26mg/ml.The expression of zinc-alpha 2-glycoprotein（ZAG）was detected by Western Blotting,respectively,after the intervention of 24h,48h and 72h respectively.Results:1).On the 7 day node,the gray level of ZAG in blank group was lower than that of model group ZAG,and the gray level of ZAG in the blank group of the 14 day model group was higher than that of the model group ZAG protein.2).Through this experiment,HUA mice and mice ZAG band gray value contrast,found that the mouse model of kidney uric acid than the blank group compared the effects of ZAG,RT-PCR results also showed that high uric acid renal ZAG mRNA levels higher than the control mice ZAG mRNA,indicating that the kidney cells secrete ZAG.3).The cell experiment showed that different concentrations of uric acid intervention at different time,ZAG had no change in HK-2 cells,the cell is different from the animal,and the induction time is limited,so the expression level of ZAG did not change 4).Uric acid may be through other means that zinc-2-glycoprotein（ZAG）change.Conclusion:1)By comparing the gray value of ZAG strips between HUA mice and blank mice,it is found that the kidney of uric acid mice can affect ZAG changes compared with the blank group.RT-PCR results also indicate that the ZAG mRNA level of high uric acid kidney is higher than that of blank mice,indicating that the renal cells secrete ZAG.At the same time,in the 7 day model mice,ZAG expression was high and ZAG mRNA expression increased,and lipid metabolism disorder,especially triglyceride decreased significantly,indicating that uric acid may cause changes in ZAG protein by affecting triglyceride metabolism.The 14 day model group of HUA mice and mice ZAG band gray value contrast,found a mouse model of ZAG is kidney uric acid lowering,but ZAGmRNA is a rising trend,while also considering the uric acid antioxidant and pro oxidant effects,through the literature,promote oxidation can lead to the production of inflammatory factors,so on the7 day and 14 day protein expression showed the opposite.2)another cell experiment showed that there was no change in ZAG in HK-2 cells at different concentrations of uric acid for different time.Because ZAG was not different from animals,and the induction time was limited,the expression level of ZAG did not change.3)uric acid may change zinc-2-glycoprotein（ZAG）through other ways.