Study Of The Mechanism Of SIRT1/CLC-3-mediated Pulsed Radiofrequency Therapy On Neuropathic Pain In SNI Rats

Objective and Research significance:The pathogenesis of neuropathic pain(NP)is unclear and it is difficult to treat it.Studies have proved that silencing information regulator 1(SIRT1)can change the plasticity of neurons by silencing the expression of related genes,thus alleviating NP.Chlorine channel 3(CLC-3)is involved in the central and peripheral sensitization of NP,but the interaction of SIRT1 and CLC-3 and the mechanism of their joint action on NP are still unclear.TNF-α is a key inflammatory factor mediating peripheral and central sensitization,but its involvement in the regulation of SIRT1 and CLC-3 in NP has not been reported.Pulsed radiofrequency(PRF)is commonly used in the treatment of NP,but the mechanism of PRF is not clear.Through the research design,the interaction between SIRT1 and CLC-3 in NP and their joint effect in NP were discussed.TNF-a was first proposed as a key inflammatory factor involved in the inverse regulation of SIRT1 and CLC-3 in NP.Finally,the specific mechanism of PRF treatment for NP was discussed by applying PRF intervention in NP model to simulate the clinical treatment process.The changes of TNF-α,SIRT1 and CLC-3 in PRF treatment and their interaction rules were further demonstrated,laying a foundation for NP basic research and clinical treatmentMethod:This experiment is mainly divided into four chapters,which are listed as belows:1.The SNI model was established in rats to observe the changes of pain behavior and its mechanical pain threshold was detected.According to Decosterd et al,ligation and injury of sciatic nerve branches were performed,and an SNI model was established.PWMT was detected preoperatively and postoperatively to observe the behavioral changes before and after modeling.2.The expressions of SIRT1,CLC-3 and IL-10 in the dorsal root ganglion(DRG)of the surgical side from L4 to L5 at different time points after surgery were detected in SD rats,and the postoperative changes and possibly mechanism of SIRT1,CLC-3 and IL-10 were observed.By intrathecal injection of NS or SRT1720(SIRT1 agonist)before and after surgery,the changes of its behavior,SIRT1,CLC-3 and other related factors were observed.Immunofluorescence staining was used to detect DRG and observe the expression and localization of SIRT1 and CLC-3 in different neurons in DRG,as well as the relationship between SIRT1 and CLC-3 in DRG.3.After surgery,the expression of TNF-a was observed in rats.And after the surgery,NS and Etanercept(TNF-a antagonist)were injected intrathecally,and the changes of their behavior and the expressions of TNF-α,SIRT1 and CLC-3 were observed.4.After surgery,RF electrodes were placed on local of the ligation of the sciatic nerve after surgery,and the behavioral changes,SIRT1 and CLC-3 expression changes in SNI rats before and after PRF treatment were compared..Results:After surgery,the PWMT of the rat was decreased obviously,and the behavior was changed obviously.After surgery,SIRT1,CLC-3 and IL-10 expression decreased,while TNF-α expression continued to increase.Intrathecal injection of SRT1720 preoperatively or postoperatively can up-regulate CLC-3 and increase the level of PWMT.Expressions of SIRT1 and CLC-3 were up-regulated and PWMT was improved after intrathecal injection of Etanercept.After PRF treatment,SIRT1 and CLC-3 expression were increased,TNF-a expression was further down-regulated,and PWMT level was gradually increased..Conclusion:SIRT1 can participate in the formation of NP by affecting the activation of neurons in DRG.Moreover,it mediates the occurrence and development of NP by acting on CLC-3.In addition,TNF-a has a reverse regulation effect on the expression of SIR1 and CLC-3,inhibiting the expression of TNF-α can up-regulate the expression of SIRT1 and CLC-3,thus alleviating NP.PRF,as a commonly used clinical intervention method for NP,can achieve the effect of NP treatment through the up-regulation of SIRT1 and CLC-3.Meanwhile,PRF may regulate the expression of TNF-α in DRG,and negatively regulate the expression of SIRT1 and CLC-3,so as to achieve the effect of NP treatment.