Abstract
Tianxiangdan(TXD),a traditional Chinese herbal remedy,demonstrates efficacy in mitigating myocardial ischemia-reperfusion(I/R)-induced damage.This study employed network phar-macology to evaluate the therapeutic targets and mechanisms of TXD in treating I/R.High-performance liquid chromatography-mass spectrometry(HPLC-MS)identified 86 com-pounds in TXD.Network pharmacological analysis predicted potential target genes and their modes of action.Cardiac function,ischaemic ST changes,lactate dehydrogenase(LDH),malondialdehyde(MDA),superoxide dismutase(SOD)activity,myocardial fiber,and infarct size were assessed using in vivo and in vitro I/R injury models.Estrogen receptor alpha(ERα)protein expression and estradiol(E2)levels were measured to confirm TXD's impact on estro-gen levels and ERα expression.To examine if TXD reduces I/R injury through ERα,an AZD group(300 nmol·L-1 AZD9496 and 15％TXD serum)was compared to a TXD group(15％TXD serum).The study hypothesized that TXD upregulates the ERα-mediated iron metamorphosis pathway.I/R injury-induced ferroptosis was identified using a Fer-1 group(1.0 μmol·L-1 Fer-1 and 15％TXD serum)to elucidate the potential association between ferroptosis and ERαproteins.A DCFH-DA probe detected reactive oxygen species(ROS)and Fe2+,while Western blotting assessed target protein expression.Both in vitro and in vivo experiments demon-strated that TXD attenuated I/R injury by reducing elevated ST-segment levels,improving cardiac injury biomarkers(LDH,MDA,and SOD),alleviating pathological features,and pre-venting I/R-induced loss of cell viability in vitro.The effects and mechanisms of TXD on I/R in-jury-associated ferroptosis were investigated using I/R-induced H9c2 cells.The TXD group showed significantly decreased ROS and Fe2+levels,while the AZ group(treated with AZD9496)exhibited increased levels.The TXD group demonstrated enhanced expression of ERα and glutathione peroxidase 4(GPX4),with reduced levels of P53 protein and ferritin-heavy polypeptide 1(FTH1).The AZ group exhibited contrasting effects on these expression levels.The literature indicated a novel connection between ERα and ferroptosis.TXD activ-ates the ERα signaling pathway,promoting protection against I/R-induced myocardial cell ferroptosis.This study provides evidence supporting TXD use for myocardial ischemia treat-ment,particularly in older female patients who may benefit from its therapeutic outcomes.
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