Explore The Effect Of Acupuncture Regulation Of MCP-1/CCR2 Pathway On Cardiac Inflammatory Injury In Spontaneously Hypertensive Rats Based On The Theory Of "Preventing Disease From Exacerbating"

Objrctive:Based on the theory of "preventing disease from exacerbating",this study investigated the mechanism of acupuncture at Renying(ST9)in early hypertension to regulate THE MCP-1 /CCR2 pathway,reduce blood pressure,delay blood pressure rise and improve cardiac inflammatory injury in spontaneously hypertensive rats.To provide experimental basis for further revealing the hypotensive mechanism of Renying(ST9)and improving the hypotensive effect of clinical acupuncture,protecting heart tissue and improving heart function.Method:Ten 4-week-old Wistar rats were used as control group.Twenty 4-week-old Spontaneously hypertensive rats rats were randomly divided into model group and acupuncture group,with 10 rats in each group.After a week of adaptive feeding,the rats in the acupuncture group were needled regularly at Renying(ST9)on both sides for 10 minutes every day,5 times a week for 24 weeks.Rats in control group and model group were not needled,but only grabbed for the same time.Systolic blood pressure was measured every 3weeks.All rats were sampled at 25 weeks and blood and heart were collected.The morphological changes of rat heart tissue were observed by HE staining and Masson staining.The contents of MCP-1,CCR2,IL-1β,IL-6 and TNF-α in serum of rats were determined by ELISA.IHC staining was used to detect the positive expression of CD68,MCP-1,CCR2,IL-1β,IL-6 and TNF-α in heart tissue.The relative expression levels of MCP-1 m RNA,CCR2 m RNA,il-1 βm RNA,il-6 m RNA and TNF-αm RNA in heart tissue were detected by QPCR.Results:1.Before acupuncture,systolic and diastolic blood pressure in model group and acupuncture group were higher than those in control group,with significant difference(P <0.01).Comparison of blood pressure changes within each group: Blood pressure in model group increased rapidly until the 6th week after acupuncture,and remained at a high level continuously,with significant differences at each time point compared with before acupuncture(P < 0.01);Systolic blood pressure of acupuncture group began 6 weeks after acupuncture,and there were significant differences between each time point and before acupuncture(P < 0.05).The diastolic pressure of acupuncture group increased slowly from the beginning of acupuncture,and there were significant differences at each time point and before acupuncture(P < 0.05).Comparison of blood pressure changes among all groups: the blood pressure of model group and acupuncture group was always higher than that of control group,with significant difference(P < 0.01);Compared with the model group,the blood pressure of the acupuncture group was lower than that of the model group from 3 weeks later,and the systolic blood pressure of the acupuncture group was significantly different from that of the model group after 3,6,9,12,15,18 and 24 weeks(P < 0.01).After 9,12,18 and 24 weeks of acupuncture,there was significant difference in diastolic blood pressure between the acupuncture group and the model group(P < 0.05).2.HE staining results showed that the central muscle fibers of the heart tissue of rats in the control group were orderly arranged,the structure of myocardial cells was intact,and the nucleoli were obvious without obvious pathological changes.Compared with the control group,the myocardial fibers in model group were disordered and irregular.Myocardial fiber rupture,myocardial fiber structure dissolution;There was more infiltration of inflammatory cells in the myocardial space.However,in the acupuncture group,myocardial fibers were arranged in a more regular way,with less broken myocardial fibers and less infiltration of inflammatory cells.3.Masson staining results showed that a small amount of collagen fibers could be seen in the myocardial space in the heart tissue of the control rats,with relatively sparse distribution and neat arrangement of muscle fibers.A large number of blue collagen fibers were observed in the myocardial space of the heart tissue of rats in model group,and the myocardial fibers were disordered and broken.Compared with the model group,the collagen fibers in myocardial interstitium in acupuncture group were slightly lower and the arrangement of myocardial fibers was more regular.4.ELISA results showed that the contents of MCP-1,CCR2,IL-1β,IL-6 and TNF-αwere lower in the control group.Compared with control group,the contents of MCP-1,CCR2,IL-1β,IL-6 and TNF-α in model group and acupuncture group were increased(P < 0.05);Compared with model group,the contents of MCP-1,CCR2,IL-1β,IL-6 and TNF-α in acupuncture group were decreased,and the contents of MCP-1,IL-1β,IL-6 and TNF-α were decreased significantly(P < 0.05).5.IHC results showed that the positive expressions of CD68 and MCP-1,CCR2,IL-1β,IL-6 and TNF-α were low in the control group.Compared with the control group,the positive expressions of CD68,Mc P-1,CCR2,IL-1β,IL-6 and TNF-α were higher in model group and acupuncture group(P < 0.05).Compared with model group,the positive expressions of CD68,MCP-1,CCR2,IL-1β and IL-6 in acupuncture group were significantly decreased(P < 0.05).6.QPCR results showed that compared with the control group,the m RNA relative expression levels of MCP-1,CCR2,IL-1β,IL-6 and TNF-α in model group were increased,and the m RNA relative expression levels of MCP-1,CCR2,IL-16 and TNF-α were significantly increased(P < 0.05).Compared with model group,the m RNA relative expression levels of MCP-1,CCR2,IL-1β and TNF-α in acupuncture group were significantly decreased(P < 0.05).Conclusion:1.Acupuncture at Renying(ST9)at the early stage of hypertension can reduce blood pressure in spontaneously hypertensive rats,and has a significant effect on delaying the continuous increase of blood pressure.2.Acupuncture at Renying(ST9)in early stage of hypertension can delay the pathological process of heart tissue.3.Acupuncture at Renying(ST9)can reduce systemic inflammatory response,improve cardiac inflammatory injury,and delay the development of cardiac inflammation.Its mechanism is related to regulating MCP-1/CCR2 pathway,reducing the infiltration of cardiac macrophages and the expression of inflammatory factors.