| ObjectiveEssential hypertension(EH)is a clinically common chronic disease with a high incidence worldwide.The continuous increase in blood pressure is the main manifestation,but its cause and mechanism have not yet been clearly stated.Once the disease is diagnosed,it is often necessary to take antihypertensive drugs for life.At present,the commonly used antihypertensive drugs,while stabilizing blood pressure,are often accompanied by many side effects,such as increasing the incidence of diabetes.Chinese medicine does not have a clear record of hypertension,but according to its common clinical manifestations such as dizziness,head bloating,and irritability,the disease can be attributed to liver-yang hyperactivity syndrome for treatment.Studies have shown that acupuncture as a part of traditional Chinese medicine can effectively lower blood pressure.Compared with Western medicine antihypertensive drugs,acupuncture treatment has almost no side effects,is simple and inexpensive to use,and can reduce the financial burden of patients with hypertension to a certain extent.During the onset of hypertension,heart,brain,and kidney damage will occur.Therefore,while treating hypertension,attention should be paid to the damage caused by the disease to the corresponding target organs,and timely measures should be taken to protect the target organs.Among them,high blood pressure and brain protection have become a close clinical concern in the contemporary era.In the research of acupuncture and antihypertensive in the previous research group,it has been concluded that acupuncture and antihypertensive can also have brain protective effects through brain histomorphology,but its specific mechanism is less studied.Current research has found that hippocampus can shrink due to elevated blood pressure,but after related therapeutic interventions,the hippocampus can play a role in reducing blood pressure through related biological pathways.Therefore,in this study,spontaneously hypertensive rats(SHR)were selected as the research object.Acupuncture was used as an intervention method.By comparing the changes of blood pressure in different groups of rats,Elisa was used to detect glucocorticoids in hippocampus and peripheral serum.Receptor(GR)expression and expression of each component of the hypothalamus-pituitary-adrenal cortex(HPA)axis.Label free and PRM proteomics were used to screen and verify protein expression in the hippocampus,and to analyze acupuncture and tonic The central hypotensive mechanism of manipulation aims to provide a new theoretical basis for acupuncture treatment of hypertension.MethodsForty-two male SHR rats were randomly divided into a model group(M),a twirling-reinforcing method group(B),and a twirling reducing method group(X),and 14 WKY rats were used as a blank control group.Each acupuncture group was treated with acupuncture and twirling reinforcing and reducing method group to intervene on both sides of Taichong.The model group and blank group were not treated with acupuncture,and were only given the corresponding fixation and treatment 20 minutes once a day.Tail artery blood pressure was measured in each group with a non-invasive blood pressure meter 1 day before acupuncture and 2,6,8,12,14days after acupuncture.After the intervention,blood was taken from the abdominal aorta,the brain was decapitated and the hippocampal tissue was separated.The morphological changes of the hippocampus were observed by HE staining,.The hippocampal glucocorticoid receptor(GR)and the Corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),and cortisol(CORT)expression were measured by Elisa method.Label free and PRM proteomics techniques were used to detect differential protein expression in the hippocampus of each group.Results1.Changes in blood pressure of rats before and after treatment in each groupCompared with the blank group,the systolic blood pressure of the tail artery in the other groups was significantly increased before and after acupuncture(P<0.01).Compared with the model group,on the 6th day of treatment,the systolic blood pressure of the twirling reducing method group was significantly reduced(P<0.05);On the 8th day of treatment,the systolic blood pressure in the twirling reinforcing method group decreased significantly(P<0.05),and the systolic pressure in the twirling reducing method group significantly decreased(P<0.01);On the 14th day of treatment,the systolic blood pressure in the twirling reinforcing group decreased significantly(P<0.01).Compared with the twirling reinforcing method group,on the 12th day of treatment,the systolic blood pressure in the twirling reducing method group decreased significantly(P<0.05);On the 14th day of treatment,the systolic blood pressure in the twirling reducing method group decreased significantly(P<0.01).2.Morphological changes of hippocampal CA1 and CA3 in each group of ratsThe HE staining results showed that the neurons in the blank group had complete structure,regular cell morphology,normal intercellular spaces,and clear nucleoli;In the model group,the staining became lighter,the neurons atrophied,the arrangement was loose,and the nuclear contraction occurred;In the twirling reinforcing method group,neurons were lost,cell morphology changed,and some cell structures were degraded;Compared with the twirling reinforcing method group,the twirling reducing method group had closer neurons,clear nucleoli,and more complete cell structure.3.Changes of GR in hippocampus and the contents of HPA axis of each group of ratsCompared with the blank group,the expression of GR in the hippocampus of the model group was significantly increased,and the difference was statistically significant(P<0.01);Compared with the model group,the expression of GR in the hippocampus of the rats in the twirling reinforcing method group and the twirling reducing method group was reduced,and the difference was statistically significant(P<0.05);Compared with the twirling reinforcing method group,the hippocampal GR expression in the twirling reducing method group was significantly reduced,and the difference was statistically significant(P<0.05).The overall results showed that the expression of GR in hippocampus of the rats in each group was in the order of small group to big group is blank group<twirling reducing method<twirling reinforcing method<model group.Compared with the blank group,the plasma ACTH,CORT,and CRH in the model group were significantly increased(P<0.01);Compared with the model group,the ACTH,CORT,and CRH of rats in the twirling reinforcing method group and twirling reducing method group decreased significantly(P<0.05,P<0.01);Compared with the twirling reinforcing method group,the ACTH and CRH in the twirling reducing method group decreased(P<0.05).4.Differential protein expression in hippocampus of each group of ratsA total of 1163 differential proteins were retrieved from the hippocampal proteomics by label-free quantitative Label Free technology.Compared with the model group,there were 314 differential proteins in the blank group,of which 123 were up-regulated and 191 were down-regulated,and 48 differential proteins were included in the supplement method,of which 25 were up-regulated and 23 were down-regulated.There are 82 differential proteins in the group,45 up-regulated and 37 down-regulated;compared with the blank group,330 differential proteins in the diarrhea group,of which 144 are up-regulated,186 down-regulated differential proteins,supplement method There were 319 differential proteins in the group,of which 132 were up-regulated and 187 were down-regulated;compared with the model group,there were 70 differential proteins,of which 33 were up-regulated and 37 were down-regulated.According to GO function and differential protein enrichment analysis of various groups,hippocampal protein function is mainly related to cellular processes,biological regulation,metabolic processes,organelles,membranes,protein complexes,and catalytic activity.KEGG function analysis of hippocampal proteins,hippocampal protein expression is closely related to the nervous system,immune system,endocrine system,substance dependence,neurodegenerative diseases,cardiovascular diseases,transportation and catabolism.The KEGG enrichment analysis of the differential proteins in each group found that the renin-angiotensin system was inhibited by the intervention of acupuncture,twisting and relieving,and it was concluded that this pathway is one of the important ways to lower blood pressure.Further,PRM verification of hippocampal renin-angiotensin-related proteins in each group of rats was performed to obtain angiotensinogen(AGT),angiotensin converting enzyme(ACE)and renin receptor(Atp6ap2)after twisting The intervention of the method of supplementing and relieving was obviously reduced.ConclusionAcupuncture at both sides of Taichong and applying twirling-reinforcing and reducing method can effectively reduce blood pressure in spontaneously hypertensive rats,and it can also protect the hippocampus.Its mechanism of action may be through acupuncture on the hippocampus-HPA axis,improving the expression of glucocorticoid receptor(GR)in the hippocampus,inhibiting the expression of the HPA axis,reducing the secretion of ACTH,CORT,and CRH in the plasma.At the same time,the protein pathways related to the regulation of blood pressure are activated to achieve a central-peripheral coordinated hypotensive effect. |
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