The Intervention Effect Of Electroacupuncture-induced MCAO/R Convalescence Rats Blood-Brain Barrier Permeation Through TNF-α/PKCβ1/P-gp Pathway

Objective This work is aiming to investigate the impact of Electroacupuncture(EA)treatment on TNF-α、PKCβ1、P-glycoprotein(P-gp)expression,and discover the correlation between EA-induced rats Blood-Brain Barrier permeation and TNF-a/PKCβ1 pathway.Methodology Firstly,the SD rats middle cerebral artery occlusion-reperfusion(MCAO/R)model is setup with Longa improved method.After four weeks recovery,they are separated randomly into five groups,such as Ischemia,EA,and imposter-EA,EA+TNF-α suppressed and Sham-operated group.Each group has three subgroups.Ischemia and sham-operated group are fixed with cloth without EA treatment.Imposter-EA is needled on Baihui(GV20),Shuigou(GV26),connected but no actual electricity.In EA group,Baihui and Shuigou are needled with continuous 2Hz 2mA electricity for 12 minutes.In EA+TNF-α suppressed group,rats are firstly injected with Etanercept through tail vein(Etanercept diluted with saline for 5 mg/ml.Each SD rat has 0.8mg/kg).Then EA group and EA+TNF-α group are intervention with EA on the same acupuncture points and configurations 30 minutes later.Finally,brain tissue are examined after all the intervention for TNF-α,PKCβ1 and Phospho-PKCβ1 express level,P-glycoprotein position express,with ELISA,Western blot and Immunohistochemistry respectively.Result 1.The rat brain TNF-α:All the other groups decreased significantly on TNF-αcomparing to Ischemia group,(P<0.05).All EA groups increased significantly on TNF-αcomparing to sham-operated group,(P<0.05).EA+TNF-α suppressed group had significantly less TNF-α than pure-EA group comparing to pure-EA group,(P<0.05).2.The rat brain PKCβ1 and Phospho-PKCβ1 expression level:comparing to Sham-operated group,Ischemia group didn’t show any significant difference on PKCβ1 expression(P>0.05),EA group showed higher ratio of Phospho-PKCβ1 and PKCβ1 compared to Ischemia group and sham-operated group and EA+TNF-α suppressed group,(P<0.05).3.P-glycoprotein positive expression and immunohistochemical scores(IHS):comparing to Sham-operated group,Ischemia group didn’t show any significant difference on P-gp expression and IHS(P>0.05),EA group had lower P-gp level and IHS comparing to lschemia group and EA+TNF-αsuppressed group,(P<0.05).Conclusion 1.2HzEA treatment can increase TNF-α in MCAO/R convalescence rats’ brain,and raise the PKCβ1 expression and activation,and decrease P-gp expression.2.The mechanism of EA-induced Rats Blood-Brain Barrier permeation has strong correlation with TNF-α/PKCβ1/P-gp pathway through decrease the P-gp to influence the efflux system.