Mechanisms Of Buyang Huanwu Decoction Influencing Connexin43 After Focal Ischemia

Objective:Gap junction protein 43(connexin43,Cx43)is one of the important channels that brain astrocytes material exchange,its expression is related to brain damage and repair after stroked.From ischemia-reperfusion rats after the process of the body's resistance to damage and repair,testing hippocampus Cx43 expression,Buyanghuanwu decoction(BYHWD)may influence the expression of Cx43 for protecting brain,intend to illustrate the role of astrocytes in traditional Chinese medicine treatment of cerebral ischemia and its mechanism which has certain innovation significance,but also reveals BYHWD which has unique advantages in the treatment of cerebral ischemia(pointedly according to the clinic condition),with modern science and technology of Chinese medicine theory to expound and prove the goals of a small step,has the positive significance for the world to better understand,accept and apply chinese medicine.Methods:By using line switch method to establish the rat middle cerebral artery embolism(middle cerebral artery occlusion,MCAO)model.1.The expression of MCAO rats hippocampal Cx43 text.Rats building,for 1 day,7 days later,using immunohistochemical method and western bolt(WB)method to detect,detect each group of Cx43 expression change.2.The influence mechanism text.BYHWD treatment 1 days later,use ELISA method to detect cytokine(interleukin 1 beta,IL-1 beta and interleukin 10,IL-10),by using the nf-kappa B agonists increase after inflammation,compare the expression of Cx43 in hippocampal CA1 area,and using HE staining observed the change of cell morphology change.3.After BYHWD treatment 7 days,immunohistochemical staining to observe the expression of bFGF,with the combination of bFGF after neutralizing antibodies,in each groups of hippocampal CA1 area,the expression of Cx43 are compared,at the same time,HE dyeing observation of cell morphology change.3.After intervention connexin43 expression ultimately also five soup against cerebral ischemia effect experiments.After BYHWD treatment 1 days,through the combination of Cx43 inhibitors,'detect hippocampal Malondialdehyde(MDA),Superoxide Dismutase(SOD),nitric oxide(Nitrogen Monoxide,NO),observe whether Cx43 is inhibited after affects by BYHWD protective effect on brain after ischemia reperfusion.Results:1.After brain ischemia 1 days,compared with control group,model group' s expression of Cx43 rise obviously,otherwisecompared with model way,the expression of Cx43 reduce in BYHWD group(P<0.05);Cerebral ischemia after 7 days compared with control model group,the expression of Cx43 raise,and compared with model group,the expression of Cx43 BYHWD group increased(P<0.05).2.Cerebral ischemia after 1 days,compared with model group,BYHWD group of cerebral ischemia reperfusion after 1 days,IL-1 beta reduced and IL-10 increased(P<0.05);With combination of the nf-kappa B inhibition BYHWD also supplement in hippocampal CA1 region and cut effect of Cx43 inhibited(P<0.05),HE staining showed the inhibit group of neuronal damage compared with BYHWD group3.the brain ischemia 7 days BYHWD also enhance the expression of bFGF and Cx43;With the Combination of bFGF neutralizing antibody,in hippocampal CA1 region,the expression of Cx43 reduce in the BYHWD group(P<0.05).HE staining showed more the neuronal damage than the BYHWD group.4.After Cerebral ischemia 1 day,compared with control group,model group MDA,NO raised,and SOD decreased(P<0.05);Compared with control group,BYHWD,the quantity of MDA,NO decreased,and SOD raised;BYHWD with Cx43 inhibitors GAP-134 cut the effect on MDA,NO,SOD after cerebral ischemia.Conclusions:1.BYHW in cerebral ischemia and reperfusion early supplement in hippocampal astrocytes inhibited Cx43 expression,compared to increasing expression of Cx43 in brain lated ischemia.2.In the early ischemia,BYHWD mediated adjusted the hippocampus the expression of Cx43 by inflammatory cytokines I1-1 beta and I1-10,acted as the brain damage resistance.In the late of ischemia,BYHWD mediated by neurotrophic factor(bFGF)adjusted the expression of Cx43 in hippocampus,and play the role of the brain repaired.3.In the early ischemia,BYHWD may chang the content of MDA,SOD and NO in brain protection by influencing the Cx43.