Target Screening And Confirmation Of Antihypertensive Effects Of Monoester Alkaloids In Aconite

Objective: Clarify the antihypertensive effect of monoester alkaloids in aconite,screen and confirm its antihypertensive target,and provide basis for elucidating the pharmacological mechanism of monoester alkaloids.Method: Using essential SHR,the changes in blood pressure of rats after the transient administration of three monoester alkaloids via the femoral vein were detected by the femoral artery invasive manometry method,and by the femoral artery invasive manometry method Changes in blood pressure of rats after instantaneous administration of BAC via gastric tube intubation;secondly,the online hypertension drug target intelligent analysis platform was used to screen out potential drug-lowering effect target proteins,and the simulation docking technology of Discovery Studio software and external surface plasma were used SPR)experiment to verify the binding of the three monoester alkaloids to the target protein;to verify the effect of monoester alkaloids on the activity of the target protein in vitro through enzyme kinetics experiments;The blood pressure changes of rats after 14 days of intragastric administration were detected by the invasive manometry method,and the serum concentrations of the relevant components of the RAAS system were determined by ELISA method.Results: Transient administration of BAC through femoral vein significantly reduced the blood pressure of SHR;when BMC was administered transiently through femoral vein,it had a slight effect on reducing the blood pressure of SHR;but when BHC was administered,the blood pressure of rats had no significant change.Using femoral artery invasive manometry to detect different doses of BAC administered via gastric tube intubation,high-dose BAC can significantly reduce the blood pressure of SHR,while low-to-medium doses have no effect on blood pressure.Some of the targets screened by the intelligent analysis system of drug targets mainly include ACE,ACE2,etc.The virtual molecular docking bioanalysis technology shows that ACE and ACE2 have a good combination with the three monoester alkaloids.SPR in vitro experiments further verified the binding effects of the three alkaloid components with the two targets.Animal in vitro enzyme kinetics experiments show that the administration of BAC can inhibit ACE enzyme activity and stimulate ACE2 enzyme activity.The blood pressure of SHR can be reduced after high-dose BAC is detected by invasive femoral artery manometry,but there is no hypotensive effect when low-dose is administered by gavage.Plasma collected after high-dose BAC gavage is tested by ELISA to determine the content of ACE Decrease,the content of ACE2 increases;there is no significant change in the content of ACE and ACE2 in the plasma collected after medium and low dose gavage.Conclusions: Monoester alkaloids in Aconite can lower the blood pressure of SHR.BAC can reduce the blood pressure of SHR by co-targeting the ACE-Ang II-AT1 axis and ACE2-Ang(1-7)-Mas axis of the RASS system.