Study On The Mechanism Of PKC Inflammatory Response Pathway Mediated By P2X3 Receptor In The Notopterygium Treatment Of "Forelimb Pain"

Objective : The purpose of this study is to explore the molecular mechanism of Notopterygium treating upper limb arthralgia from the level of P2X3 receptor-mediated PKC inflammatory response pathway,clarify its scientific connotation of clinical treatment of upper limb arthralgia,and provide experimental basis for Notopterygium clinical application.Methods:1 Using the MTT method,in vitro experiments were conducted to study the proliferation of rat fibroblast synoviocytes(FLS)at different concentrations(150,175,200,225,250 ?g / mL)and different intervention times(24,48,72h)Impact;The growth-synchronized FLS was divided into control group,notopterygium group,and positive group,and the expressions of P2X3,Rab7,PKC,CXCL1,TLR4 gene and protein were detected by RT-PCR and Western-Blot method.2 Randomly divide the rats into a blank group,a model group,a positive control group,and a notopterygium group,and replicate the rat upper extremity arthritis model with Freund's adjuvant.Each group begins to administer the drug after the model is successful,and the drug is given by continuous intragastric administration for 21 days After removing the joint cavity.The expressions of P2X3,Rab7,PKC,CXCL1,TLR4 gene and protein were detected by RT-PCR method and Western-Blot method,respectively.Results:1 The MTT method determines the Notopterygium administration time is 24 h,and the concentration is 175?g / mL.Notopterygium can significantly inhibit the expression of P2X3 receptor mRNA and protein in FLS(P <0.05 or P <0.01)and the expression of its transporter Rab7 mRNA and protein(P <0.05 or P <0.01);significantly inhibit downstream PKC mRNA and protein Expression(P <0.05),and down-regulate the expression of related inflammatory factors TLR4 and CXCL1 gene and protein(P <0.05 or P <0.01);significantly reduce the expression of NF-?B mRNA and protein under PKC pathway(P <0.05 or P <0.01).2 Notopterygium can significantly inhibit the expression of P2X3 receptor mRNA and protein in the joint cavity of adjuvant upper extremity arthritis model(P <0.05)and the expression of its transporter Rab7 mRNA and protein(P <0.05);significantly inhibit the downstream PKC mRNA and protein Expression(P <0.05),and down-regulate the expression of related inflammatory factors TLR4 and CXCL1 gene and protein(P <0.05 or P <0.01);significantly reduce the expression of NF-?B mRNA and protein under the PKC pathway(P<0.01).Conclusion : The mechanism of Notopterygium in treating upper limb arthralgia is related to the inhibition of P2X3 receptor-mediated PKC inflammation pathway.