Study On The Confirmation Of Active Ingredients And Multi-level Mechanism Of Action Of Centella Asiatica(L.) Urban In Anti-cardiac Hypertrophy Based On Systems Pharmacology

Pathological cardiac hypertrophy(CH)is an incurable heart disease,leading to an increased risk of heart failure due to the lack of safe and effective strategies.Traditional Chinese Medicine,with its efficacy and low toxicity,is increasingly taking great advantage in the treatment of various chronic diseases.Centella asiatica(L.)Urb.(CA),a traditional medicinal and natural plant,has been reported to be effective in the treatment of cardiovascular diseases,but its efficacy and potential mechanisms in alleviating CH have not been fully investigated.Therefore,based on a systems pharmacology approach and in vivo and in vitro experiments,this paper explored the therapeutic effects of CA and the most promising active compounds,and clarified the mechanism of action of CA for CH treatment.Methods:(1)The CH model was constructed using subcutaneous injection of isoprenaline(ISO)in C57BL/6 mice to investigate the anti-cardiac hypertrophy activity of CA.(2)A system pharmacology approach and a CH mouse model were used to screen for potential active ingredients and potential targets of CA,construct a compound-target network and assay mouse cardiac function parameters,thereby exploring the major active ingredients of CA for CH treatment.(3)Gene enrichment and KEGG pathway analysis were employed to reveal the molecular mechanism of action of CA.(4)Based on the HL-1 mouse model of cardiomyocyte oxidative damage,the multi-level mechanisms of action of active compound in CA for CH treatment was validated using Western Blot,kits and RT-q PCR as well as section staining.Results:(1)Combined with the parameters of cardiac function and molecular markers of cardiac injury and hypertrophy in mice,CA improved heart rate,reduced heart weight-to-body weight ratio,reversed abnormal changes in the levels of BNP,ANP,AST,TNT and LDH,and alleviated CH.(2)13 active ingredients and 195 corresponding targets were screened from CA by ADME screening model and target identification.Next,based on target analysis and evaluation in mouse models,it was found that AA significantly improved cardiac function parameters and abnormal changes in cardiomyocytes,being the main active compound of CA for CH treatment.(3)Functional enrichment analysis showed that CA was mainly involved in PI3K/AKT,HIF1,MAPK and RAGE signaling pathways to synergistically regulate several biological processes to exert cardioprotective effects.(4)In vivo and in vitro experiments verified that AA synergistically regulates YAP/PI3K/AKT axis and NF-κB signaling pathway,which are mainly involved in anti-inflammatory,anti-apoptotic and antioxidant biological processes to exert anti-cardiac hypertrophy effects.Conclusion: Overall,the multi-level mechanism of action of CA for CH was demonstrated by a systems pharmacological approach,which provides a paradigm for systematically deciphering the mechanism of action of natural plants for disease treatment and offers new ideas for the development of new drugs.