YiQiFuMai Powder Injection Attenuates Ischemia/Reperfusion-Induced Myocardial Apoptosis Through AMPK Activation

Abstract

The YiQiFuMai powder injection (YQFM), a traditional Chinese medicine (TCM) prescription re-developed based on the well-known TCM formula Sheng-maisan, showed a wide range of pharmacological activities in cardiovascular diseases in clinics. However, its role in protection against myocardial ischemia/reperfusion (MI/R) injury has not been elucidated. The present study not only evaluated the cardioprotective effect of YQFM from MI/R injury but also investigated the potential molecular mechanisms both in vivo and in vitro. The myocardium infarct size, production of lactate dehydrogenase (LDH), creatine kinase (CK), cardiac function, TUNEL staining, and caspase-3 activity were measured. Cell viability was determined, and cell apoptosis was measured by Hoechst 33342 staining and flow cytometry. Mitochondrial membrane potential (ΔΨm) was measured, and ATP content was quantified by bioluminescent assay. Expression of apoptosis-related proteins, including Caspase-3, Bcl-2, Bax, AMPKα, and phospho-AMPKα, was analyzed by western blotting. AMPKα siRNA transfection was also applied to the mechanism elucidation. YQFM at a concentration of 1.06 g/kg significantly reduced myocardium infarct size and the production of LDH, CK in serum, improved the cardiac function, and also produced a significant decrease of apoptotic index. Further, combined treatment with compound C partly attenuated the anti-apoptotic effect of YQFM. In addition, pretreatment with YQFM ranging from 25 to 400 μg/mL markedly improved cell viability and decreased LDH release. Moreover, YQFM inhibited H9c2 apoptosis, blocked the expression of caspase-3, and modulated Bcl-2 and Bax proteins, leading to an increased mitochondrial membrane potential and cellular ATP content. Mechanistically, YQFM activated AMP-activated protein kinase (AMPK) signaling pathways whereas pretreatment with AMPK inhibitor Compound C and application of transfection with AMPKα siRNA attenuated the anti-apoptotic effect of YQFM. Our results indicated that YQFM could provide significant cardioprotection against MI/R injury, and potential mechanisms might suppress cardiomyocytes apoptosis, at least in part, through activating the AMPK signaling pathways.

Cardioprotection by combination of three compounds from ShengMai preparations in mice with myocardial ischemia/reperfusion injury through AMPK activation-mediated mitochondrial fission

“…Consistent with our previous study10, the present study demonstrated the protective effect of Shengmai preparations YQFM on MI/R injury. Similar to YQFM, the active ingredients combination GRS also exerted cardioprotective effect with the same potency, as shown by improved LV function, decreased infarct size, cell viability loss and LDH activities.…”

“…At the end of the experiment, blood was collected and serum was separated by centrifugation, the LDH was detected as previously described10. After collecting the blood samples, the hearts were removed.…”

“…In brief, the cells were transfected with 80 nmol/L of total siRNA by using Transfection Reagent as previously described10. To evaluated the efficiency of the AMPK siRNA, total cell lysates were subjected to SDS-PAGE for immunoblotting analysis with β-actin as a reference.…”

“…Plenty literatures have reported that AMPK is cardioprotective during MI/R via promoting glucose uptake and glucose transporter translocation, inhibiting apoptosis and decreasing myocardial infarction789. In our previous study, we found that the YiQiFuMai powder injection, a traditional Chinese medicine prescription re-developed based on the well-known TCM formula Sheng-maisan, could provide significant cardioprotection against MI/R injury, and potential mechanisms might to suppression of cardiomyocytes apoptosis at least in part through activating the AMPK signaling pathways10. Recently, regulation of the cytoplasmic dynamin-related protein 1 (Drp1) emerges as a potential molecular target by which AMPK activation inhibits mitochondrial dysfunction and alleviates endoplasmic reticulum stress-associated endothelial dysfunction11.…”

Extract of Sheng-Mai-San Ameliorates Myocardial Ischemia-Induced Heart Failure by Modulating Ca2+-Calcineurin-Mediated Drp1 Signaling Pathways

“…Treatments consisting of multiple drugs directed against multiple pathways and targets have stronger therapeutic efficacy for complex multifactorial diseases [ 7 , 8 ]. The effective evidence from clinical trials contributes to the acceptance of TCM can be useful in the treatment of cardiovascular diseases [ 9 , 10 , 11 ].…”