OBJECTIVE To investigate the mechanism of action of Quzhuo Tongbi Granules(QZTB)in reducing uric acid and anti-inflammation in mice with hyperuricemia(HUA).METHODS The components of QZTB were identified by ultra-performance liquid chromatography-mass spectrometry(UPLC-MS).Sixty-four C57BL/6 mice were randomly divided into control group,model group,benzbromarone group,QZTB low-dose group,QZTB medium-dose group,QZTB high-dose group,MCC950 group,and MCC950+QZTB medium-dose group,with 8 mice in each group.Adenine(100 mg·kg-1)and potassium oxonate(500 mg·kg-1)were used to establish the HUA mouse model.Except for the control group,all other groups underwent 2 weeks of modeling followed by 4 weeks of treatment.After 2 weeks of modeling,blood was collected from the orbital venous plexus to measure serum uric acid(SUA)levels as the criterion for successful model induction.Mice were sacrificed after 4 weeks of continuous treatment for sample collection.An automatic biochemical analyzer was used to measure serum levels of SUA,urea nitrogen(BUN),and creatinine(Cr).Enzyme-linked immunosorbent assay(ELISA)was employed to detect serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-18(IL-18),and tumor necrosis factor-α(TNF-α).The qPCR was used to assess mRNA expression of urate transporter 1(URAT1),ATP-binding cassette transporter G2(ABCG2),glucose transporter 9(GLUT9),and PDZ domain-contai-ning protein kinase 1(PDZK1)in kidney tissue.Western blot was performed to measure protein expression of urate transporters(URAT1,ABCG2,GLUT9,PDZK1),nuclear transcription factor κB(NF-κB)total protein,phosphorylated NF-κB(p-NF-κB),Nod-like receptor protein 3(NLRP3),Cleaved Caspase-1 and Pro-Caspase-1 proteins in kidney tissue.Immunohistochemistry was used to determine the expression levels of urate transporters(URAT1,ABCG2,PDZK1,GLUT9)in kidney tissue.RESULTS A to-tal of 9 representative active ingredients were identified in QZTB.Two weeks after modeling,SUA in the model group was significantly increased compared with that in the control group(P<0.000 1).Four weeks after administration,serum SUA,BUN and Cr in the model group were significantly increased(P<0.000 1),IL-1β,IL-6,IL-18 and TNF-α levels were increased(P<0.01,P<0.001),the expression of ABCG2 and PDZK1 proteins in renal tissue was decreased(P<0.01,P<0.001,P<0.000 1),and the ex-pression of URAT1,GLUT9,NLRP3,p-NF-κB p65/NF-κB p65 and Cleaved Caspase-1/Pro-Caspase-1 proteins was significantly increased(P<0.01,P<0.001,P<0.000 1).Compared with the model group,SUA,BUN and Cr in the benzbromarone group and the low-,medium-and high-dose QZTB intervention groups were reduced to varying degrees(P<0.001,P<0.000 1).QZTB could ef-fectively reduce the levels of serum inflammatory factors IL-1β,IL-6,IL-18 and TNF-α(P<0.05,P<0.01),increase the expres-sion of ABCG2 and PDZK1 proteins in renal tissue(P<0.05,P<0.01,P<0.000 1),and downregulate the expression of URAT1,GLUT9,NLRP3,p-NF-κB p65/NF-κB p65 and Cleaved Caspase-1/Pro-Caspase-1 proteins(P<0.05,P<0.01,P<0.001,P<0.000 1).Compared with the model group,the MCC950 group downregulated the protein expressions of NLRP3,p-NF-κB p65/NF-κB p65,and Cleaved Caspase-1/Pro-Caspase-1(P<0.01).Compared with the MCC950 group or the QZTB group,the MCC950+QZTB group downregulated the protein expressions of NLRP3,p-NF-κB p65/NF-κB p65,and Cleaved Caspase-1/Pro-Caspase-1(P<0.05,P<0.01,P<0.000 1).CONCLUSION QZTB can promote uric acid excretion by inhibiting the NF-κB/NLRP3 signaling pathway,thereby improving the symptoms of HUA.

OBJECTIVE To investigate the mechanism of action of Quzhuo Tongbi Granules(QZTB)in reducing uric acid and anti-inflammation in mice with hyperuricemia(HUA).METHODS The components of QZTB were identified by ultra-performance liquid chromatography-mass spectrometry(UPLC-MS).Sixty-four C57BL/6 mice were randomly divided into control group,model group,benzbromarone group,QZTB low-dose group,QZTB medium-dose group,QZTB high-dose group,MCC950 group,and MCC950+QZTB medium-dose group,with 8 mice in each group.Adenine(100 mg·kg-1)and potassium oxonate(500 mg·kg-1)were used to establish the HUA mouse model.Except for the control group,all other groups underwent 2 weeks of modeling followed by 4 weeks of treatment.After 2 weeks of modeling,blood was collected from the orbital venous plexus to measure serum uric acid(SUA)levels as the criterion for successful model induction.Mice were sacrificed after 4 weeks of continuous treatment for sample collection.An automatic biochemical analyzer was used to measure serum levels of SUA,urea nitrogen(BUN),and creatinine(Cr).Enzyme-linked immunosorbent assay(ELISA)was employed to detect serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-18(IL-18),and tumor necrosis factor-α(TNF-α).The qPCR was used to assess mRNA expression of urate transporter 1(URAT1),ATP-binding cassette transporter G2(ABCG2),glucose transporter 9(GLUT9),and PDZ domain-contai-ning protein kinase 1(PDZK1)in kidney tissue.Western blot was performed to measure protein expression of urate transporters(URAT1,ABCG2,GLUT9,PDZK1),nuclear transcription factor κB(NF-κB)total protein,phosphorylated NF-κB(p-NF-κB),Nod-like receptor protein 3(NLRP3),Cleaved Caspase-1 and Pro-Caspase-1 proteins in kidney tissue.Immunohistochemistry was used to determine the expression levels of urate transporters(URAT1,ABCG2,PDZK1,GLUT9)in kidney tissue.RESULTS A to-tal of 9 representative active ingredients were identified in QZTB.Two weeks after modeling,SUA in the model group was significantly increased compared with that in the control group(P<0.000 1).Four weeks after administration,serum SUA,BUN and Cr in the model group were significantly increased(P<0.000 1),IL-1β,IL-6,IL-18 and TNF-α levels were increased(P<0.01,P<0.001),the expression of ABCG2 and PDZK1 proteins in renal tissue was decreased(P<0.01,P<0.001,P<0.000 1),and the ex-pression of URAT1,GLUT9,NLRP3,p-NF-κB p65/NF-κB p65 and Cleaved Caspase-1/Pro-Caspase-1 proteins was significantly increased(P<0.01,P<0.001,P<0.000 1).Compared with the model group,SUA,BUN and Cr in the benzbromarone group and the low-,medium-and high-dose QZTB intervention groups were reduced to varying degrees(P<0.001,P<0.000 1).QZTB could ef-fectively reduce the levels of serum inflammatory factors IL-1β,IL-6,IL-18 and TNF-α(P<0.05,P<0.01),increase the expres-sion of ABCG2 and PDZK1 proteins in renal tissue(P<0.05,P<0.01,P<0.000 1),and downregulate the expression of URAT1,GLUT9,NLRP3,p-NF-κB p65/NF-κB p65 and Cleaved Caspase-1/Pro-Caspase-1 proteins(P<0.05,P<0.01,P<0.001,P<0.000 1).Compared with the model group,the MCC950 group downregulated the protein expressions of NLRP3,p-NF-κB p65/NF-κB p65,and Cleaved Caspase-1/Pro-Caspase-1(P<0.01).Compared with the MCC950 group or the QZTB group,the MCC950+QZTB group downregulated the protein expressions of NLRP3,p-NF-κB p65/NF-κB p65,and Cleaved Caspase-1/Pro-Caspase-1(P<0.05,P<0.01,P<0.000 1).CONCLUSION QZTB can promote uric acid excretion by inhibiting the NF-κB/NLRP3 signaling pathway,thereby improving the symptoms of HUA.