The Efficacy And Mechanism Study On Winqing Compatibility Of Cinnamaldehyde And Polydatin In The Treatment Of Hyperuricemia

Objective: Based on the characteristics of gout attack and the theory of compatibility of traditional Chinese medicine prescriptions,this study intends to explore the efficacy and mechanism of Wen Qing compatibility and Qing Wen compatibility of Cinnamaldehyde and polydatin(Guihu)in the treatment of hyperuricemia by using modern pharmacology,molecular biology and metabonomics technology.To explore whether evening medication of Wen Qing combination of Cinnamaldehyde and polydatin can better interfere with hyperuricemia which expect to prevent the nocturnal attack of gout.Methods: 120 SPF male Kunming mice were randomly divided into10 normal mice and 110 model mice according to their weight.The laboratory fixed light(L): Dark(D)(L: D=12:12,dark period is 08:00 am to 20:00 pm,light period is 20:00 pm to 08:00 am).The model mice were administrated orally by gavage of 200 mg/kg potassium oxyzinate and 200mg/kg hypoxanthine(modeling drug)at 9 am with once a day for 7 days to establish hyperuricemia model of mice,while the normal mice were administrated orally by gavage of 0.5% sodium carboxymethylcellulose(CMC-Na).Two hours after administration of the 7th day,the blood was collected from the orbital venous plexus of all the mice to determine serum uric acid(SUA)level.90 successful model micewere divided into model group,febustat group,benzbromarone group,Wenqing compatibility of cinnamaldehyde and polydatin low dose group(WQ-L group),Wenqing compatibility middle dose group(WQ-M group),Wenqing compatibility high dose group(WQ-H group),Qingwen compatibility of polydatin and cinnamaldehyde group(Qingwen group),polydatin group and cinnamaldehyde group(n=10 mice/group).One hour after administration of the modeling drug,each group was administrated orally by gavage of corresponding therapeutic drugs once a day for 28 days,while the normal mice and the model mice were administrated orally by gavage of 0.5%sodium carboxymethylcellulose(CMC-Na).On the 27 th day of treatment,the urine was collected in a metabolic cage.One hour after treatment of the28 th day,the mice were sacrificed after blood was drawn from eyeball and the kidneys and livers were taken to reserve.The levels of SUA,serum creatinine(SCr),blood usea nitrogen(BUN),blood adenosine deaminase(ADA),urine uric acid(UUA),urine creatinine(UCr)and the activities of blood aspartate aminotrans-ferase(GOT),glutamic-pyruvic transaminase(GPT),and the activity of liver xanthine oxidase(XOD)were measured using commercial kits assay reagent kit.Pathological sections of the kidney were made to observe the renal injury.The expression levels of urate-anion transporter 1(URAT1),Glucose transporter 9(GLUT9),Organic anion transporters 1(OAT1)m RNA in kidney and ADA,circadian locomotor output cycles kaput(CLOCK),brain and muscle arnt-like protein 1(BMAL1)m RNA in liver were measured by q PCR.The expression levels of URAT1,GLUT9,OAT1 proteininin in kidney and ADA,XOD,CLOCK,BMAL1 proteininin in liver were determined by western blotting.Established the metabolic fingerprints of serum and urine in hyperuricemia mice by 1H-NMR technique.Clarified the difference of metabolic fingerprints between groups by the supervised method orthogonal partial least-squares discriminant analysis(OPLS-DA).Variable importance in the projection(VIP)and t test were used to screened the potential biomarkers associated with hyperuricemia from serum and urine,and then compared the changes of their levels.Results: Compared with the normal group,the level of SUA in the mice before treatment increased significantly(P < 0.01).Compared with the model group after administration,the levels of SUA of mice in all treatment groups decreased significantly(P<0.01,P<0.05);the levels of SCr in WQ-H group decreased significantly(P<0.05);the levels of BUN and the activity of XOD reduced significantly(P<0.01,P<0.05)and the levels of UUA and UCr increased significantly in all treatment groups except the WQ-L group(P<0.01,P<0.05);the activity of ADA in all treatment groups except the WQ-L group and polydatin group reduced significantly(P < 0.01,P < 0.05);the activity of GPT and GOT in all treatment groups were no significant difference.All treatment groups could improve renal damage of hyperuricemia mice.Compared with the model group,all treatment groups could significantly decrease the m RNA and protein expression levels of URAT1,GLUT9 in kidney and ADA in liver(P<0.01,P<0.05);the protein expression levels of XOD in liver reduced significantly in all treatment groups(P < 0.01);the m RNA and protein expression levels of OAT1 in kidney and BMAL1 in liver increased significantly in all treatment groups(P < 0.01,P < 0.05);the m RNA expression levels of CLOCK in liver increased significantly in all treatment groups(P < 0.01,P < 0.05);the protein expression levels of CLOCK in liver increased significantly in WQ-H group(P<0.05).WQ-H group could significantly reverse 7 and 3 potential biomarkers related to hyperuricemia in serum and urine,respectively(P<0.01,P<0.05).Qingwen group could significantly reverse 5 and 3 potential biomarkers related to hyperuricemia in serum and urine,respectively(P<0.01,P<0.05).Conclusion: Both the monomer group and the compatibility group of Cinnamaldehyde and polydatin have clear anti-hyperuricemia effect.Compared with the Qingwen group,polydatin group and cinnamaldehyde group,Wenqing compatibility group had better uric acid lowering effect.The mechanism is related to the inhibition of XOD and ADA activities in the liver,and the regulation of Uric acid transporter in kidney and the regulation of intestinal microflora metabolism,purine metabolism,amino acid metabolism,lipid metabolism and glucose metabolism.In addition,it is possible to interfere with hyperuricemia by regulating liver clock genes.