Regulation Of QishenYiQi Formula And Its Components Of YiQi And HuoXue On Vasodilation In Hypertensive Rats

Qi Shen Yi Qi is composed of astragalus,salvia miltiorrhiza,panax notoginseng and incense,with invigorating Qi for consolidating superficies and activating blood effect of removing stasis,it can be applied to a variety of cardiovascular diseases.The dysfunction of vasodilation and contraction is related to the occurrence and development of many cardiovascular diseases,the most important of which is primary hypertension.In the state of disease,vasodilation response is weakened,vasospastic contraction,thus accelerating the development of hypertension,and even cause myocardial infarction,cerebrovascular accident and other serious complications.According to modern pharmacological research,QSYQ can significantly increase myocardial contractility,reduce myocardial fibrosis,improve myocardial energy metabolism,and have good prevention and treatment effect on myocardial ischemia and myocardial ischemia reperfusion injury.Although widely used clinically,however,due to the complexity of compound Chinese medicine ingredients,and characterized by multi-component,multi-target and multi-pathway,Its mechanism of regulating vascular function is still unclear,there are few studies on vascular dependent relaxation.Therefore,it has become the focus of our research to explore the effects of QSYQ and its components on vasodilation and contraction function.Objective:This study is to selecting the Qi Shen Yi Qi extract（QSYQ）,Yi Qi components-huangqi extract（YQ）,Huo Xue components-salvia miltiorrhiza,panax notoginseng extract（HX）as research object and using the Power Lab physiological recording system and Radnoti organ perfusion system,with endothelial integrity and endothelial vascular ring removal as the model,then to record the contraction or relaxation curves in normal SD rat thoracic aorta.To observe the effect of QSYQ and its components on vascular tension,and to investigate the diastolic mechanism of presystolic vascular ring.Furthermore,the effects of QSYQ on vascular endothelial function and its protective effect in spontaneous hypertensive rats were studied.Network pharmacology was used to analyze the important targets and possible pathways of QSYQ.From the tissue level to the animal level system to explore the mechanism of regulating the vascular function of QSYQ,and QSYQ prescription is better than the Yi Qi components,Huo Xue components.Methods:Establish the rat thoracic aortic ring model in vitro,systematically evaluate the effects of QSYQ,YQ and HX on NE preconstriction vascular ring with and without endothelial integrity in normal rats;And select the most appropriate concentration of drugs for the following study.The nitric oxide synthase inhibitor（L-NAME）and cycloxygenase inhibitor（INDO）were pretreated the endothelial complete vascular ring to observe the effect of QSYQ,YQ,HX on the vascular tension of the endothelial integrity.Calcium activated potassium channel blocker（TEA）,adenosine triphosphate sensitive potassium channel blocker（Glib）,inward rectifying potassium channel blocker（Ba Cl₂）and voltage activated potassium channel blocker（4-AP）were pretreated to remove endothelial vascular ring,and the effects of different potassium channels on the vasodilatory activity of QSYQ were observed.And then,spontaneous hypertension rats（SHR）were used as the model,and normal Wistar rats（WKY）were used as the control group.SHR was divided into 5 groups.After 7 days of pre-administration,take out the aorta and divide it into two parts;The model of thoracic aortic ring in isolated rats was adopted;Observe the effects of QSYQ,YQ and HX on NE preconstriction vascular ring with endothelial integrity.The aorta was removed for pathological section,H&E staining and Masson staining were used to evaluate the protective effect of QSYQ on blood vessels;Immunohistochemical staining was used to evaluate the expression of proteins related to vasoconstrictor function.Finally,we used network pharmacology analysis.Through literature mining,we established a database on hypertension and conducted core analysis,establish a"disease-pathway"network to find the most relevant pathway for primary hypertension;Analyzed the mechanism involved in vasodilation,obtained several key targets;The"component-target"diagram was obtained by analyzing the relationship between the active components in QSYQ and the important targets of vasodilation caused by hypertension.Results:In the isolated rat model of thoracic aortic ring,0.2mg/m L QSYQ,0.1mg/m L YQ and HX could relax thoracic aortic vessels with or without intact endothelium in normal rats.In the case of intact endothelium,the relaxation effect was stronger,and the relaxation effect of QSYQ was better than that of YQ and HX（p<0.001）;Only L-NAME pretreatment of complete vascular ring of endothelium could significantly inhibit the diastolic effect of QSYQ and other components（p<0.001）,but INDO do not.Pretreatment with Ba Cl₂ inhibitor to remove endothelial vascular ring can block QSYQ-induced vasodilatory（p<0.05）,the other three blockers had no significant inhibitory effect.One week after preadministration,QSYQ treated SHR vasodilation improved（p<0.01）.However,the vasodilation effect of SHR treated by YQ and HX did not improve.H&E staining showed that compared with WKY group,the proportion of inner/outer diameter of blood vessels in SHR without administration was smaller（p<0.001）.QSYQ treated SHR thoracic aorta vessel diameter/outside diameter ratio was significantly amplified（p<0.05）,the thickness of the vessel walls is also decreasing（p<0.01）.Masson staining showed that the collagen deposition area and fibrosis in the wall of SHR vessels treated with QSYQ tended to decrease.IHC detected the expression levels of e NOS antibody and AKT antibody to vasodilatation-related proteins after QSYQ treatment of SHR,and compared with the expression levels of untreated SHR（p<0.01）,and are a slightly highher activity.The e NOS pathway has the highest correlation with essential hypertension.To view the"vasodilation"pathway in the mechanism of hypertension,and it is a key target for vasodilation that the targets of NO,e NOS and AKT are in a core position.It can be seen from the“component-target”diagram that the active components contained in QSYQ are related to e NOS and NO targets in vasodilation.Conclusion:The results of our study found that 0.2mg/m L QSYQ could effectively relax the blood vessels of normal rats and hypertensive rats,and the effect of vasodilation was better than that of YQ and HX.After one week of pre-administration,QSYQ can improve the vascular function of hypertensive rats,reconstruct blood vessels,reduce vascular fibrosis and protect cardiovascular function.When the endothelium is intact,the vasodilation mainly functions through the AKT-e NOS-NO pathway.Smooth muscle-mediated vasodilation in endothelial incompleteness may be related to the opening of the K_IR.In addition,QSYQ is better than YQand HX in vasodilation and improvement of vascular endothelial function,which is helpful for further understanding of TCM component and provides experimental basis for compatibility theory of TCM component.