P2Y6 Receptor-mediated JAK/STAT Signaling Pathway Involved In Neuropathic Pain Mechanism

Objective:1.To observe the change of P2Y6 receptor and JAK/STAT signaling pathway in neuropathic pain?NPP?of spinal dorsal horn ganglion,and its relationship with NPP;2.To explore the P2Y6 receptor in spinal dorsal horn ganglion With the regulation of JAK/STAT signaling pathway,we further explored the specific pathogenesis of spinal cord dorsal ganglion P2Y6receptors involved in JAP/STAT signaling pathway in NPP.Methods:1.80 male SD rats,each about 220²50g,for the study.According to the principle of complete random number table,they were divided into 4 groups:sham group?S/control group,n=20?,neuropathic pain group?C/model group,n=20?,and P2Y6receptor inhibitor MRS2578 group.?M/inhibition group,n=20?and P2Y6 receptor agonist UDP group?U/activated group,n=20?.2.S group:only the right sciatic nerve trunk was isolated without ligation;group C was isolated from the right sciatic nerve and ligated;M group and U group were injected into the abdominal cavity with P2Y6 receptor inhibitors on the basis of group C model.1 ml each of MRS2578?concentration:10?m/ml?and P2Y6 receptor agonist UDP?100?m/ml?.In the S group and the C group,1 ml normal saline was injected daily into the experimental rat with a 1 ml empty needle.The abdominal cavity.Each group was subdivided into four subgroups?reported preoperatively?T₀?,postoperative 3d?T₁?,postoperative 7d?T₂?,postoperative 14d?T₃?,n=5 per subgroup?.3.Rats'mechanical side pain threshold?MWT?and thermal pain threshold?TWL?were measured at T₀,T₁,T₂,and T₃ in each group.4.The behaviors of each group were measured at T₀,T₁,T₂,and T_3.After each index was sacrificed,each rat was sacrificed and L4-5 spinal cord tissue was removed.The expression of SOCS3,p-JAK2,and p-STAT3 proteins and the expression of P2Y6R mRNA,STAT3mRNA,JAK2 mRNA,and SOCS3 mRNA were detected by Western blot and RT-PCR.5.After each group was anesthetized at T₀,T₁,T₂,and T₃,the thorax was dissected,and about 2 ml of blood was collected at the apex of the heartbeat.Centrifugation was performed at a low speed.The serum was carefully taken with a pipette and the ELISA method was used to detect the changes of IL-6concentration in the serum.Results:1).MWT:C group was lower at T₂ and T₃than S group?P<0.05?;at T₂ and T₃,U group was lower than group C?P<0.05?;at T₂ and T₃,group M was higher than C.The group increased?P<0.05?.TWL:C group was lower at T₂ and T₃ than S group?P<0.05?;at T₂ and T₃,U group was lower than group C?P<0.05?;at T₂ and T₃,group M was higher than group C?P<0.05?.2).Changes of serum IL-6 concentration:C and U groups began to increase at T₂,and it was more obvious at T₃ than at T₂ in the same group.C group was higher than S group at T₂ and T₃.U group was significantly higher than C group at T₂ and T₃.Compared with group C,group M was significantly lower at T₂and T₃,P<0.05.3).Four groups of rat spinal cord L4-5 The expression of P2Y6mRNA,JAK2 mRNA,STAT3 mRNA and SOCS3 mRNA in the tissue:C and U groups began to increase at T₂,which was different from T₀ and T₁.Compared with S group,C group increased at T₂ and T₃?P<0.05?.At T₂ and T₃,the U group was significantly higher than the C group?P<0.05?,while the M group was lower than the C group?P<0.05?,while the S group was changed at the T₂ and T₃?P>0.05?.4).Expression of p-JAK2,p-STAT3 and SOCS3 protein in the four groups of L4-5 tissue:C group and U group began to increase at T₂,which was different from T₀ and T₁;group C was at T₂ and T₃ compared with S group.It was elevated?P<0.05?;at T₂,T₃,U group was significantly higher than that of C group?P<0.05?,while M group was significantly lower than C group?P<0.05?.Conclusion:1.Separation and ligation of the sciatic nerve successfully established the CCI model,which enabled the activation of P2Y6 receptor in the spinal dorsal horn ganglion.2.Pathogenesis of NPP may be related to activation of the JAK/STAT signaling pathway;3.The inhibition of spinal dorsal horn ganglion P2Y6 receptors,pain can alleviate mental derangement rational mechanism may be related to inhibit the JAK/STAT signal transduction pathway,cut the JAK/STAT signal transduction pathway related mRNA and protein expression and of IL-6 in plasma generation and release.