Based On PKA/CREB Signal Pathway To Probe The Effect Of The Method Of Removing Phlegm And Removing Stasis And Accompany With Opening The Orifices On Vascular Dementia Rats

Objective Vascular dementia(VD)is the second leading cause of dementia,phlegm and blood stasis is an important pathological factor for the onset of VD,due to the special physiologic function and structure of brain,need at the same time by using the method of removing phlegm and blood stasis accompany with opening the orifices.This article reviews and summarizes the understanding of ancient doctors on VD disease,etiology,pathogenesis,treatment and prescription,and summarizes the progress of modern Chinese medicine in the treatment of VD.To investigate the mechanis m and treatment principle of the pathogenesis of VD from removing phlegm and blood stasis accompany with opening the orifices therapy,we use the atherosclerosis composite cerebral ischemia SD rat as animal VD model,Objective To observe the effects of the method of removing phlegm and blood stasis accompany and opening the orifices therapy on behavior,blood lipid,inflammatory factor,microvessel density in hippocampal cortex,hippocampal brain histopathology and PKA-CREB signaling path way in rats.To explore the effect and mechanism of the method of removing phlegm and blood stasis accompany with opening the orifices therapy on the model,and to provide theoretical basis and experimental basis for the prevention and cure of VD by removing phlegm and blood stasis accompany with aromatic resuscitation therapy.Method: 1.Theoretical exploration: Collating related literatures related to the therapy of removing phlegm and blood stasis accompany with opening the orifices of TCM,analyzing the concept and connotation of the removing phlegm and blood stasis accompany with opening the orifices therapy,and discussing its theory and clinical foundation for prevention and treatment of VD.2.Experimental research:We use the atherosclerotic accompany with cerebral ischemia model of SD rats as VD animal model,taking PKA-CREB signaling pathway as the target,to study the mechanism of the effect of removing phlegm and blood stasis accompany with opening the orifices therapy on VD.Useying the effects of the Morris water maze on the behavior of rats to detect the learning and memory ability of the rats.Useing HE staining and immunohistochemical staining to observe the microvascular density(MVD)in the hippocampus and the pathological changes in the hippocampus of rats.Useing ELISA assay to determine the exp ression of inflammatory factors?vascular endothelial growth factor and related substances in the upper and lower levels of PKA-CREB pathway in the hippocampus of rats.Useing the Western blot method to detect the main protein expression of the PKA-CREB pathway in the hippocam pus of rats.Result: 1.MORRIS water maze Directional navigation test: After 4 weeks,comparison with the Blank group,the Model group rats found the platform time is prolonged obviously(P<0.01),the swimming distance was significantly increased(P<0.01);Compared with the Model group,each drug group rats found platform time was shortened(P<0.05,P<0.01),swimming distance were reduced(P<0.05,P<0.01);Compared with Ditan Decoction group,The Huoxue grou p found platform time was prolonged(P>0.05),total swimming distance was significantly increased(P<0.01).Compared with the Ditan Decoction group,the time of Flavored Ditan Decoction group was shorter(P>0.05),and the total distance of swimming was shortened(P<0.05).The spatial exploration experiment shows a similar trend to the directional navigation experiment.2.The effect on blood lipid and inflammatory factors Compared with the Blank group,the blood lipid in the model group increased significantly(P<0.01).4 weeks after administration of each drug group,the levels of TC,TG and LDL were reduced to different degrees,and HDL increased to varying degrees,of which the most significant group was Flavored Ditan Decoction group.For inflammatory factors,the expression of inflammatory factors in hippocampal tissue of Model group was significantly higher than that in Blank group(P<0.01).After treatment,inflammatory factors in hippocampus tissue of rats in each group were reduced to some extent,among which the most significant group was Flavored Ditan Decoction.3.The effect of on the microvascular density and the pathomorphology of the hippocampus.Related tests after four weeks of drug use,The content of Ang-1 and VEGF in the hippocampus of Model group rats was significantly lower than that in the Blank group(p<0.01);Eatch treatment group compare d with Model group,Ang-1 and VEGF content in hippocampus of rats were significantly increased(p<0.01),the Flavored Ditan Decoction group increased the most;Immunity immunohistochemistry showed that the number of microvessels in the hippocampus of the Model group was significantly less than that in the blank group(P<0.01),after drug use The vascular density of the hippocampal cortex in each group had a different degree increase,among which Flavored Ditan Decoction group was the most obvious,followed by Ditan Decoction.HE staining showed that: A large number of vertebral cells were found in the CA1 area of the Blank group,closely arranged,neat.Model group rats CA1 area cones lost a lot,residual cell swelling,residual cell swelling,Cell interstitial sparsity.Comp arison of each drug group with the model group,The degree of vertebral cell loss was lighter,cell degeneration and necrosis less,which Flavored Ditan Decoction group was most obvious.4.Effect of on PKA-CREB signaling pathway in VD model rats The expression levels of cAMP,PKA,CREB,NGF and BDNF in hippocampus of Model group were significantly lower than those in Blank group.After treatment,all above indexes in each drug group increased in different degrees.Flavored Ditan Decoction group increased significantly.Conclusion: 1.Phlegm and blood stasis are the basic factors that lead to the onset of VD,removing phlegm and blood stasis accompany with opening the or ifices therapy is an important method to prevent and cure VD effectively.2.The method of removing phlegm and blood stasis accompany with opening the orifices therapy improve the cognitive function and the ability of autonomic activity in VD rats.3.The method of removing phlegm and blood stasis accompany with opening the orifices therapy in the treatment of VD is related to lowering the blood lipid and inflammatory factors,promoting angiogenesis,increasing the expression of related substances in PKA-CREB signaling pathway,and exerting anti ischemia and brain cell apoptosis.