| Objective:This study intends to investigate the mechanism on effcets of Polydatin-cinnamaldehyde compatibility on hyperuricemia and its renal protective effect based on molecular biology techniques and metabolomics techniques.The aim is to provide some reference for the treatment of hyperuricemia in traditional Chinese medicine group.Methods:A total of 60 adult male SD rats were randomly divided to three groups according to their body weight(n=10 rats/group):the control group,the hyperuricemia model group,the Allopurinol group,and the Polydatin-Cinnamaldehyde(PC)low dose group,PC medium dose group,and PC high dose group.Except the control group,rats of the other groups were subcutaneously injected with 200 mg/kg potassium oxonate,after 7 hours the rats were intragastrically administered 100 mg/kg hypoxanthine and 250mg/kg ethambutol once daily for 26 days to establish hyperuricemia model of rat.Before 6 hours from subcutaneous administration on the 5th day,the blood was collected from the orbital venous plexus of all the rats to determine serum uric acid(SUA)level before administration.On the 6th day,the rats except the control group and model group were administrated orally by gavage drugs with once a day for 21 days.24-hour urine samples and 24-hour fecal samples from each rat were collected from metabolism cages on day 25.After 6 hours from subcutaneous administration on the 26th day,the rats were sacrificed after blood was collected from the abdominal aorta and the kidneys were taken to reserve.The levels of SUA,blood usea nitrogen(BUN),and creatinine(Cr)were measured using commercial kits assay reagent kit.Kidney injure molecule-1(KIM-1),neutrophil gelatinase-associated lipocalin,urine microalbumin(NGAL),and 24 hour urine microalbumin(U-mALb)levels in urine of rats were measured by enzyme-linked immuno sorbent assay(ELISA).The expression levels of angiotensin?(AngII)and cyclooxygenase-2(COX-2)mRNA in kidney were measured by qPCR.The expression levels of AngII and COX-2 proteininin in kidney were determined by Western blotting.The expression levels of TGF-?1 and?-SMA protein in kidneys were measured by immunohistochemistry.The ~1H-NMR technique was used to establish the metabolic fingerprints of serum,urine and feces in hyperuricemia rats.The supervised method orthogonal partial least-squares discriminant analysis(OPLS-DA)models were established to clarify the difference between groups.The potential biomarkers associated with hyperuricemia from serum,urine and feces were screened by variable importance in the projection(VIP)and t test,then the changes in the levels of selected biomarkers were compared.Results:Compared with the normal group,the level of SUA in the rats before treatment increased significantly(P<0.01).After administration,the levels of SUA of rats in the PC medium dose group and the PC high dose group decreased significantly(P<0.01,P<0.05),and the levels of BUN of rats decreased significantly in PC medium dose group and the PC high dose group compared with the model group(P<0.01,P<0.05).All of three doses of PC groups could significantly reduce the levels of KIM-1 NGAL and 24 h U-mALb compared with the model group(P<0.01,P<0.05).The results of renal histopathology showed that PC medium dose group and PC high dose group could significantly improve the renal injury in hyperuricemia rats compared with the model group(P<0.01,P<0.05),and the degree of renal injury from rats in PC medium dose group was the least.The expression levels of COX-2 mRNA in kidney of rats in the middle dose group and the high dose group of PC decreased significantly compared with the model group(P<0.01,P<0.05).Compared with the model group,all of three doses of PC groups could significantly reduce the expression levels of AngII mRNA in the kidney of rats(P<0.01,P<0.05),and significantly reduce the expression levels of AngII and COX-2 protein in the kidney of rats(P<0.01).The protein expression levels of TGF-?1 and?-SMA in kidney of rats were significantly decreased in three doses of PC groups compared with the model group(P<0.01),and the expression levels of TGF-?1 and?-SMA protein in the kidney of rats from PC medium dose group were the lowest.PC could significantly reverse 12,6 and 5 potential biomarkers related to hyperuricemia in serum,urine and feces,respectively(P<0.05),which could effectively interfere with metabolic disorders in serum,urine and feces of rats with hyperuricemia.Conclusion:The combination of polydatin and cinnamaldehyde has obvious anti-hyperuricemia and renal protection effect.The mechanism may be by inhibiting the expressions of AngII and COX-2 mRNA in kidney of rats,and inhibiting the expressions of AngII,COX-2,TGF-?1 and?-SMA protein in kidney of rats,so as to improve renal injury caused by hyperuricemia.In the meantime,PC may effectively interfere with hyperuricemia in rats by regulating glucose metabolism,lipid metabolism,amino acid metabolism,purine metabolism,intestinal microflora metabolism,and renal excretion function in rats with hyperuricemia. |
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