The Mechanism Study Of Electroacupuncturing Jiaji Points For Analgesia Based On ATP-P2X Mediated Microglia In Spinal Cord

Objective:To observe the time effect regulation of electroacupuncture(EA)at Jiaji acupoint on the protein expression of purinergic recepter P2X,ligand-gated ion channel 4(P2X4R),purinergic recepter P2X,ligand-gated ion channel 7(P2X7)and clone name of monoclonal antibody of complement receptor type 3(OX42)in chronic constriction injury(CCI)rats to investigate the mechanism of electroacupuncturing jiaji points for analgesia based on ATP-P2X mediated microglia in Spinal cord in neuropathic pain and provide theoretical basis for clinical application.Methods:After the adult male SD(Sprague-Dawley)rats were fed for 7 days,one hundred and eight SD rats were screened for thermal pain latency and divided into blank group,immediately after EA treatment group,0.5h after EA treatment group,1h after EA treatment group,2h after EA treatment group,4h after EA treatment group,12h after EA treatment group,24h after EA treatment group and model group.according to random number table,twelve rats in each group.Six rats in each group were randomly selected and there was no significant difference in the thermal pain latency(P>0.05).CCI model was established in EA treatment group and model group.The rats in the immediately,0.5h,1h,2h,4h,12h,24h after EA treatment group were treated with EA at L3?5 jiaji acupoint on both sides for 20 minutes on the 7th day after modeling.Each group was redetermined with thermal pain latency after modeling 7 days and before rats were sacrificed.Spinal cord bulging were taken,Western Blot method was used to measure the expression of P2X4,P2X7 and the immunohistochemical method was used to measure the expression of P2X4,P2X7 and OX42 in spinal dorsal horn.Results:After 7 days of modeling,the thermal pain latency of the EA treatment group and model group decreased significantly(P<0.001)compared with the blank group;After intervention,the thermal pain latency of the immediately,0.5h,1h,2h after EA treatment group and blank group were significantly higher than that of the model group(immediately and 2h after EA treatment group P<0.05,0.5h after EA treatment group P<0.01,1h after EA treatment group and blank group P<0.001),but that of the 4h,12h and 24h after EA treatment group was not significantly higher(P>0.05).Western Blot:the protein expression of P2X4 level in the immediately,0.5h,1h and 2h after EA treatment group did not increase significantly(P>0.05)compared with that in the blank group,but that in the 4h,12h,24h after EA treatment group and model group increased significantly(4h after EA treatment group P<0.01,12h,24h after EA treatment group and model group P<0.001).The protein expression of P2X4 level in the immediately,0.5h,1h and 2h after EA treatment group decreased significantly(P<0.01)compared with that in the model group,but that in the 4h,12h,24h after EA treatment group did not decrease significantly(P>0.05).The protein expression of P2X7 level in the immediately,0.5h,lh and 2h after EA treatment group did not increase significantly(P>0.05)compared with that in the blank group,but that in the 4h,12h,24h after EA treatment group and model group increased significantly(4h after EA treatment group P<0.01,12h,24h after EA treatment group and model group P<0.001).The protein expression of P2X7 level in the immediately,0.5h,lh and 2h after EA treatment group decreased significantly(immediately and 2h after EA treatment group P<0.01,0.5h and 1h after EA treatment group P<0.001)compared with that in the model group,but that in the 4h,12h and 24h after EA treatment group did not decrease significantly(P>0.05).Immunohistochemical:the protein expression of P2X4 level in the immediately,0.5h,1h and 2h after EA treatment group did not increase significantly(P>0.05)compared with that in the blank group,but that in the 4h,12h,24h after EA treatment group and model group increased significantly(P<0.001).The protein expression of P2X4 level in the immediately,0.5h,1h and 2h after EA treatment group decreased significantly(P<0.001)compared with that in the model group,but that in the 4h,12h and 24h after EA treatment group did not decrease significantly(P>0.05).The protein expression of P2X7 level in the immediately,0.5h,1h and 2h after EA treatment group did not increase significantly(P>0.05)compared with that in the blank group,but that in the 4h,12h and 24h after EA treatment group and model group increased significantly(P<0.001).The protein expression of P2X7 level in the immediately,0.5h,1h and 2h after EA treatment group decreased significantly(P<0.001)compared with that in the model group,but that in the 4h,12h and 24h after EA treatment group did not decrease significantly(P>0.05).The protein expression of OX42 level in the immediately,0.5h,1h and 2h after EA treatment group did not increase significantly(P>0.05)compared with that in the blank group,but that in the 4h,12h,24h after EA treatment group and model group increased significantly(P<0.001).The protein expression of OX42 level in the immediately,0.5h,lh and 2h after EA treatment group decreased significantly(P<0.001)compared with that in the model group,but that in the 4h,12h and 24h after EA treatment group did not decrease significantly(P>0.05).Conclusion:EA might inhibit microglial activation and decrease the protein expression of P2X4,P2X7 receptors in CCI rats to increase the thermal pain latency.The after effect can be extended for two hours or more than two hours and the peak period is about 1h after EA treatment,with a certain time effect regulation.