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The cardioprotective effects of acteoside in myocardial ischemia reperfusion injury and the underlying mechanism

BMC Cardiovascular Disorders volume 25, Article number: 329 (2025) Cite this article

Abstract

Introduction

We observed the cardioprotective effects of Acteoside (AC) on myocardial ischemia reperfusion injury (MIRI) and discussed the possible mechanisms.

Methods

Before MIRI model was established successfully, AC was administrated to SD rats by gastric route for 7 d. Punctuate paw withdrawal threshold (PWT) was recorded to reflect the pain threshold. Blood samples were collected to measure the levels of oxidative stress, myocardial enzymes and Norepinephrine (NE). Hematoxylin and eosin (HE) staining was performed to observe the pathological changes of myocardial tissues. Apoptosis of myocardial cell was determined by transferase-mediated dUTP nick end labeling (TUNEL) assay, and the expressions of Bcl-2 and Bax were determined by Western blotting. Using network pharmacological analysis, the PI3K/Akt signaling pathway was screened to be associated with both AC and MIRI. Subsequently, the expressions of PI3K, p-Akt and caspase-3 were detected by immunochemistry in myocardial tissues.

Results

We found that pre-administration of AC improved pain threshold and pathological change of myocardial structure caused by MIRI. AC reduced serum levels of myocardial enzymes and NE in MIRI. Compared with the Sham group, rats in MIRI group showed enhanced oxidative stress levels. These changes were partly reversed by AC. In addition, AC inhibited apoptosis, regulated the expression of apoptosis-related proteins. Immunochemistry analysis confirmed that AC increased the expressions of PI3K and p-Akt in myocardial tissue.

Conclusion

The cardioprotective effects of AC in MIRI were related with pain alleviation, oxidative stress, apoptosis and sympathetic nerve activity inhibition, the PI3K/Akt signal pathway activation.

Clinical trial number

Not applicable.

Peer Review reports

Introduction

According to the World Health Organization (WHO) (2021), cardiovascular diseases (CVDs), including myocardial infarction (MI), are the number one cause of death globally, accounting for an estimated 17.9 million deaths each year, which represents 32% of all global deaths [1, 2]. MI accounts for a significant proportion of these cases and cause severe complications that significantly impair quality of life and lead to long-term disability [3]. Restoring coronary blood flow as soon as feasible is the goal of MI therapy to protect myocardial tissue and function. Myocardial ischemia and reperfusion injury (MIRI) is the term used to describe how reperfusion can negate the positive effects of cardiac reflow and cause secondary myocardial injury, which leads to further cell death, an increase in the infarct region, and even heart failure [4]. MIRI has grown in importance as a factor in determining how well patients respond to reperfusion therapy. However, even with scientific and medical advancements, there is still no proven cure for MIRI.

Acteoside (AC), also known as Verbascoside, belongs to the class of phenylethanol glycosides and is naturally occurring in various medicinal plants, such as Echinacea and Cistanche [5, 6]. It has been demonstrated that AC exhibits no cytotoxic, genotoxic, and phototoxic properties even at relatively high concentrations, and is safe for oral administration in both animal and human diets [7, 8]. AC has emerged as a promising lead compound for clinical translation, demonstrating antitumor efficacy in diverse preclinical models [6, 9]. Beyond its antitumor potential, AC’s clinical rationale is strengthen by pleiotropic pharmacological properties, including antioxidant activity, anti-inflammatory effects, neuroprotection, cardiometabolic benefits, and hepatoprotection [10, 11]. These multifaceted actions, coupled with favorable pharmacokinetic profiles in recent trials, position AC as a versatile therapeutic candidate warranting rigorous clinical validation [12]. Moreover, AC has a protective effect on the cardiovascular system by preventing platelet aggregation in individuals with cardiovascular risk factors, decreasing myocardial cell apoptosis, repairing mitochondrial alterations induced by sepsis and suppressing the expression of angiotensin converting enzyme (ACE) in rats that develop spontaneous hypertension [13, 14].

The present study aimed to observe the effect of AC on pain threshold, serum levels of creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH) and cardiac troponin l (cTnl), as well as sympathetic nerve activity and pathological changes in the myocardial tissue of MIRI rats. Additional evidence was also provided regarding the impact of AC on oxidative stress and myocardial cell apoptosis. Furthermore, network pharmacology and immunohistochemistry analyses were used to verify that the cardioprotective effects of AC were associated with the enhanced expressions of PI3K and p-Akt.

Materials and methods

Animals and drug intervention

30 male SPF SD rats, weighing 230–250 g (10–12 weeks), were purchased from the Experimental Animal Center of Harbin Medical University. After adaptive feeding under controlled laboratory conditions for 7 d, the SD rats were randomly divided into the Sham, MIRI and AC + MIRI groups. The rats in AC + MIRI group were treated with AC (80 mg/kg/d) by the gastric route for 7 d, the rats in the Sham group and MIRI group received the equal volume of saline. The experimental protocol is shown in Fig. 1. AC was obtained from Jiangsu Yongjian Pharmaceutical Co., Ltd (61276-17-3).

Fig. 1
figure 1

The experimental protocol of the study. The MIRI rat model was estblished via ligation of LAD coronary artery for 30 min followed by 120 min reperfusion. In order to observe the effect of AC to MIRI, a total of 30 rats were evenly categorized into three groups: Sham group, MIRI group and AC + MIRI group (intragastric administration of 80 mg/kg/d AC for 7d before MIRI model establishment). The Sham and MIRI groups were pretreated with saline for the same period by intragastric administration

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Establishment of the MIRI rat model

The MIRI rat model was established through thoracotomy and ligation of the left anterior descending (LAD) coronary artery under anesthesia with 2% isoflurane [15]. The rats in the Sham group underwent the same thoracotomy but without LAD coronary artery ligation. The development of MI was verified when elevated ST segment and/or T waves appeared in the lead II ECG recorded by the BL-420 Biological Signal Acquisition and Processing System (Chengdu TME Technology Co., Ltd.). After the LAD coronary artery was blocked for 30 min, the blood flow recovery was 120 min.

Behavioral observation

The behavioral test, punctuate paw withdrawal threshold (PWT), was performed according to the methods of Segelcke and Luans [16, 17]. PWT was considered to be an indicator of mechanical pain threshold.

Biochemistry detection

The serum contents of CK-MB, NE and cTnl were determined by ELISA according to the manufacturer’s instructions (Nanjing Jiancheng Bioengineering Institute, E006-1-1, H096-1, 194-1-1). The LDH, superoxide dismutase (SOD) and malondialdehyde (MDA) levels in the serum were detected in line with the instructions of the kits (Nanjing Jiancheng Bioengineering Institute, A020-1-2, A001-2, A003-1-2).

Hematoxylin-eosin (HE) staining

Myocardial tissue was carefully harvested from the SD rats for HE staining. The myocardial tissue was fixed in 4% paraformaldehyde solution, processed by graded alcohols and xylene and embedded in paraffin, then transverse 2-µm-thick sections of tissue were cut and stained. The morphological study of the myocardial tissue was performed under the light microscope.

Terminal deoxynucleotidyl-transferase-mediated dUTP Nick end labeling (TUNEL) assay

Apoptotic myocardial cells were evaluated by TUNEL assay. The TUNEL staining procedure (paraffin-slides) was conducted according to the manufacturer’s instructions (Wuhan Servicebio Technology Co., Ltd, C1091). Briefly, the sections were washed with PBS following dewaxing and protease K repair. Subsequently, the appropriate amount of TUNEL reaction mixture was dropped onto the sample and incubated in the dark for 1 h at 37℃. After DAPI staining, the sections were sealed and observed under a fluorescence microscope (DMI 4000; Leica Microsystems GmbH). TUNEL-positive cells emitted green fluorescence, while DAPI stained the cell nucleus with blue fluorescence. Image J software (version 1.51n; National Institutes of Health) was employed to assess and compare the relative fluorescence intensity of TUNEL staining across the different groups.

Western blotting assay

The total protein of the myocardial tissue was extracted by RIPA lysis solution. The supernatant was collected and boiled to denature the protein. A BCA kit (Beyotime Institute of Biotechnology, P00110S) was adopted to measure the protein concentration of the samples. Equal quantities (20 µg) of protein were separated by 10-12.5% SDS-PAGE (Beyotime Institute of Biotechnology, P0015F)and transferred to polyvinylidene fluoride membrane. The membrane was blocked and sealed and then incubated with primary antibodies against Bax (1:2,000; cat. no. BA0315-2-Ig; Wuhan Boster, Biological Technology, Ltd.), Bcl-2 (1:2,000; cat. no. 12789-1-AP; Wuhan Boster, Biological Technology, Ltd.), and β-actin (1:5,000; cat. 81115-1-RR; Proteintech) overnight at 4°C. Next, the membrane was incubated with the secondary antibody labeled with HRP(1:10,000; cat. no. AS014; ABclonal, Biotech Co., Ltd.) for 1 h at 37℃. The results were observed with a gel imaging System (5200 Multi; Tanon Science and Technology Co., Ltd.) and analyzed by ImageJ software (version 1.51n).

Network pharmacology analysis

Screening MIRI-related genes and predicting the potential targets

MIRI pathogenic targets (restricted to Homo sapiens) were collected from the GeneCards (https://www.genecards.org/) and DisGeNET (https://www.disgenet.org/) databases. The two-dimensional structure of AC was searched in PubChem database (https://pubchem.ncbi.nlm.nih.gov). The SEA (https://sea.bkslab.org/) database was used to obtain the predicted targets for AC. The UniProt database (https://www.uniprot.org/) was applied for standardization of the gene names.

Predicting the targets of AC in MIRI

An online drawing tool (https://hiplotacademic.com) was used to plot a Venn diagram, in which the overlapping genes were the potential therapeutic targets of AC for MIRI. The protein-protein interaction (PPI) networks of the common targets were searched in the STRING 11.0 database (https://string-db.org/), restricted to H. sapiens. Cytoscape (version 3.7.0) was used to identify the key genes. The key genes were input into the DAVID database for Gene Ontology (GO) functional analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The results of the GO analysis were plotted against the KEGG bubble map, and the key targets and signaling pathways of AC in MIRI were further analyzed.

Immunohistochemistry

The expressions of PI3K(1:500; cat. no. GB11769; Wuhan Servicebio Technology Co., Ltd.) p-Akt (1:200; cat. no. GB150002; Wuhan Servicebio Technology Co., Ltd.) and caspase-3 (1:500; cat. no. GB11532; Wuhan Servicebio Technology Co., Ltd.) in myocardial tissue were examined by immunohistochemistry. The procedures regarding the immunohistochemical labeling, including dewaxing, antigen repair, blocking endogenous peroxidase activity, close serum, incubation with primary and secondary antibodies, DAB color development and staining nuclei, have been described previously. After dehydration and mounting, the results were interpreted under a light microscope and analyzed by Image J software (version 1.51n).

Statistical analysis

The results are presented as the mean ± standard deviation and were analyzed by SPSS19.0 software. One-way ANOVA followed by Tukey’s tests was used to estimate the significance of the measured parameters among the groups in the present study. P < 0.05 was considered indicate a statistically significant difference.

Results

MIRI rat model was successfully established based on ECG detection

Lead II ECG was recorded by the BL-420 Biological Signal Acquisition and Processing System. The ECG before LAD coronary artery ligation was normal in each group and ST segment elevation appeared after LAD coronary artery ligation in the MIRI group. (Fig. 2A-C).

Fig. 2
figure 2

The establishment of MIRI rat model, and AC preconditioning alleviated the pain threshold of MIRI rats. (A) The rat in the Sham group underwent thoracotomy without LAD coronary artery ligation. (B) The rat in MIRI group underwent thoracotomy and LAD coronary artery ligation. (C) Representative ECG in each group. (D) The pain threshold of each group. Results were expressed as mean ± standard deviation (n = 10). **P < 0.01; ***P < 0.001

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AC improves the pain threshold in MIRI rats

As shown in Fig. 2D, the MIRI rats had the lowest mechanical pain threshold among the three groups. The mechanical pain thresholds in the AC + MIRI group were improved compared with MIRI group, and the difference was statistically significant.

AC ameliorates myocardial injury and inhibits oxidative stress in MIRI rats

The serum levels of CK-MB, LDH and cTnl were higher in the MIRI group compared with the Sham group. Pre-administration of AC notably decreased the serum levels of CK-MB, LDH and cTnI compared with the MIRI group (Fig. 3A, B and D). These results indicated that AC could mitigate myocardial injury in MIRI rats. Compared with the Sham group, rats in the MIRI group had enhanced MDA levels and declined SOD levels (Fig. 3E and F). Furthermore, the NE serum level was higher in the MIRI group than in the Sham group (Fig. 3C). These changes were notably prevented by pre-administration of AC.

Fig. 3
figure 3

AC attenuated MIRI and inhibited oxidative stress in rats. (A) Serum level of CK-MB. (B) Serum level of cTnl. (C) Serum level of NE. (D) Serum level of LDH. (E) Serum level of MDA. (F) Serum level of SOD. Data was expressed as mean ± standard deviation (n = 10). *P < 0.05; **P < 0.01; *** P < 0.001. (F) Representative sections for HE staining were used to evaluate the pathological changes of myocardial tissue. Magnification×200, scale bar,100 μm (n = 5)

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AC improves the pathological changes in the myocardial tissues of MIRI rats

As shown in Fig. 3G, rats in the Sham group had normal myocardial structures. MIRI notably resulted in pathological features of myocardial destruction such as irregular myocardial bundles, ruptured muscle fibers, inflammatory infiltration and even necrosis. These pathological changes were ameliorated by pre-administration of AC.

AC inhibits the apoptosis of myocardial cells in MIRI rats

The results demonstrated that there was a significant increase in the relative TUNEL fluorescence ratio in the MIRI group compared with the Sham group. However, the TUNEL relative fluorescence ratio was lower in the AC + MIRI group than in the MIRI group, which indicated AC ameliorated the apoptosis of myocardial cells (Fig. 4A and B).

Fig. 4
figure 4

AC attenuated MIRI by inhibiting apoptosis, and regulting the expressions of Bcl-2, Bax and caspase-3 in myocardial tissue. (A) Representative TUNEL assay sections of myocardial tissue in each group. Magnification×200, scale bar, 100 μm. (B) The relative fluorescence ratio of TUNEL in each group was quantified using Image J. (C) Representative immunohistochemistry sections of caspase-3 expression in myocardial tissue of each group. Magnification×200, scale bar:100 μm; Magnification×400, scale bar:50 μm. (D) The expression of caspase-3 in each group was illustrated by statistical histograms. (E) The protein expressions of Bcl-2 and Bax in myocardial tissue were presented by representative Western blot bands. (F and G) The protein expressions of Bcl-2 and Bax in each group were illustrated by statistical histograms. Data was expressed as mean ± standard deviation (n = 5). *P < 0.05; **P < 0.01; ***P < 0.001

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AC regulates the expressions of apoptosis-related proteins in the myocardial tissues of MIRI rats

The expression levels of apoptosis-related proteins Bcl-2 and Bax, in myocardial tissues were analyzed by Western blotting (Fig. 4E-G). In addition, the expression of caspase-3 was analyzed by immunohistochemistry (Fig. 4C and D). The data showed that the expression levels of Bax and caspase-3 were significantly higher in the MIRI group than in the Sham group. Meanwhile, decreased expression of the anti-apoptotic protein, Bcl-2, was observed in the MIRI group compared with the Sham group. Changes in the expression of these apoptosis-related proteins in the MIRI group were significantly ameliorated in the AC + MIRI group.

Network pharmacology analysis

Predicting the targets of AC against MIRI

In total, 35 targets of AC and 2,875 targets in MIRI were retrieved from the databases. A total of 16 targets were selected as the potential targets of AC against MIRI (Fig. 5A).

Fig. 5
figure 5

Network pharmacology analysis screened and predicted the potential targets of AC against MIRI. (A) Venn diagram identified the intersecting genes of AC and MIRI.(B and C) Construction of PPI network to illustrate the interaction of the intersecting genes and screen the key targets. (D) GO analyzed the intersecting genes of AC and MIRI. (E) KEGG analyzed the pathway of intersecting genes related both AC and MIRI

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Identifying the main targets of AC against MIRI

A PPI network was constructed to identify potential interactions among the 16 targets (Fig. 5B and C). GO enrichment analysis of the key potential therapeutic targets was showed in Fig. 5D. KEGG enrichment analysis was also performed to investigate the potential signaling pathways of AC against MIRI. The PI3K/Akt signaling pathway had a high gene enrichment (Fig. 5E).

AC enhances the expressions of PI3K and p-Akt in the myocardial tissue of MIRI rats

Immunohistochemical analysis revealed the concurrent expression of PI3K and p-Akt in both the left and right ventricles. Elevated expression of PI3K and p-Akt was observed in the myocardial tissue samples from the MIRI group compared with the Sham group. Pre-administration of AC further amplified the myocardial expression levels of PI3K and p-Akt in the MIRI group, as illustrated in Fig. 6A-F.

Fig. 6
figure 6

AC enhanced the expressions of PI3K and p-Akt in myocardial tissue during MIRI. (A) Representative immunohistochemistry sections of PI3K expression in left ventricle and right ventricle. Magnification×200, scale bar:100 μm; Magnification×400, scale bar:50 μm. (B and C) The expressions of PI3K in left ventricle and right ventricle were illustrated by statistical histograms. (D) Representative immunohistochemistry sections of p-Akt expression in left ventricle and right ventricle. Magnification×200, scale bar, 100 μm; Magnification×400, scale bar, 50 μm. (E and F) The expressions of p-Akt in left ventricle and right ventricle were illustrated by statistical histograms. Data was expressed as mean ± standard deviation (n = 5). *P < 0.05; **P < 0.01; *** P < 0.001

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Discussion

MIRI is a complex phenomenon that threatens human health and life during the treatment of cardiovascular diseases. Effort has been made to explore the molecular and cellular mechanisms of MIRI and post-ischemic remodeling. The underlying mechanisms of MIRI are complex, and systematic networks are involved, including rapid changes in pH, oxidative stress, intracellular Ca2+ overload, mitochondrial dysfunction, apoptosis, endoplasmic reticulum stress, autophagy and altered metabolism [18, 19]. Evidence has suggested that AC has a cardiovascular protective effect by inhibiting ACE expression, reducing blood pressure in spontaneously hypertensive rats and ameliorating inflammation and oxidative stress to enhance cardiac function in mice and the H9c2 cardiomyocyte cells [13, 20].

Here, the influence of AC on MIRI in a rat model was observed. HE staining indicated that AC pre-administration partially prevented the myocardial destruction induced by MIRI. Clinically, CK-MB, LDH and cTnl are recognized as specific biomarkers in the diagnosis and evaluation of myocardial injury. In the present study, it was observed that the CK-MB, LDH and cTnl serum levels were significantly enhanced in MIRI rats, which were notably decreased in the AC + MIRI group. Meanwhile, the MDA level was increased in the myocardial tissue of MIRI rats accompanied by a decrease in the SOD level. As is well-known, oxidative stress serves a key role in the structural and functional alterations of myocardial cells in the pathogenesis of MIRI [21, 22]. Cellular structures, particularly cellular and mitochondrial membranes, are directly assaulted by oxidation products, resulting in membrane rupture, mitochondrial impairment and the ultimate initiation of cell apoptosis [23]. Moreover, mitochondrial dysfunction exacerbates the generation of oxidation products, notably free radicals, which in turn elicits oxidative stress responses [24]. In the present study, pre-administration of AC significantly attenuated the oxidative stress level to exert a protective effect on the heart. Similarity, AC was confirmed to attenuate oxidative stress in rats with focal cerebral ischemia reperfusion injury [25]. Serum NE measurement provides a useful method to evaluate sympathetic nerve activity [26]. The results of the study showed that AC decreased the serum level of NE, but the underlying mechanism needs further investigation. Based on its biological activities, which include anti-inflammation, antioxidant and neuroprotective effects, we consider that AC may indirectly affect the secretion and metabolism of NE by suppressing oxidative stress and the inflammatory responses in MIRI.

Emerging evidence indicates that apoptosis is one of the most important pathological mechanisms involved in the complex system networks of MIRI [27, 28]. Mitochondrial dysfunction and oxidative stress collaboratively induce the apoptosis of myocardial cells by activating apoptosis-related genes and influencing intracellular signaling transduction pathways [29]. Myocardial cells have minimal regenerative capacity. Therefore, improving cell viability and reducing infarct size have emerged as important methods to limit MIRI and the main predictors of patient prognosis. In the present study, TUNEL assay showed that there was a significant increase in apoptosis in the MIRI group compared with the Sham group. However, the rats in the AC + MIRI group exhibited a decrease in apoptosis. To further explore the impact of AC on apoptosis, the levels of the apoptosis-related targets, Bax and Bcl-2, in myocardial tissue were detected by western blotting, and the expression of caspase-3 was detected by immunohistochemistry. The results demonstrated that AC had a favorable effect in ameliorating MIRI, which was closely associated with its antioxidant and anti-apoptosis properties as described in previous studies [30, 31].

PI3K is a cytoplasmic lipid and serine/threonine kinases, and activation of PI3K directly phosphorylates its downstream target, Akt, which is a central signaling messenger in this signaling pathway. Akt actives a large number of downstream targets, including mammalian target of rapamycin (mTOR), and nuclear factor signaling erythroid 2-related factor 2 (NRF-2) [32, 33]. Recent evidence has demonstrated that activation of the PI3K/Akt signaling pathway lightens the negative influence of MIRI by synergistic augmentation of antioxidant properties, inhibition of cardiomyocyte apoptosis and inflammation, maintaining the integrity of mitochondria and promoting cell survival [34]. It has been reported that the PI3K/Akt signaling cascades are closely associated with other pathways, such as NF-κB and JNK, to mediate the development of MIRI [35]. In addition, activity of PI3K/Akt signaling pathway has been proven to be related to ischemic pre-conditioning and ischemic post-conditioning [36]. AC interacts with the PI3K/Akt pathway to influence various cellular processes, including autophagy, apoptosis, and cellular metabolism, with implications for the treatment of diseases such as diabetic kidney disease, glaucoma, osteoporosis, and possibly cancer [37, 38]. These effects are mediated through the modulation of key proteins and pathways, highlighting the multifaceted role of AC in cellular signaling. For instance, Li et al. [39] showed that AC derived from Cistanche promotes bone health by activating the PI3K/Akt/mTOR pathway, thereby mitigating dexamethasone-induced osteoporosis in vivo and in vitro. Furthermore, AC has been found to alleviate the loss of retinal ganglion cells and oxidative stress by upregulating caveolin 1, which subsequently activates the PI3K/Akt signaling pathway [40]. By targeting the PI3K/AKT/mTOR pathways, AC effectively prevents autophagy-induced apoptosis in retinal ganglion cells, offering a promising therapeutic strategy for glaucoma patients, either as a monotherapy or in combination with other autophagy inhibitors [41]. According to network pharmacological analysis in the present study, we identified the PI3K/Akt signaling pathway as the main target of AC against MIRI. Subsequent immunohistochemistry confirmed that the PI3K and p-Akt expression levels were upregulated in the AC + MIRI group compared with the MIRI group which indicated that AC ameliorated MIRI by activating the PI3K/Akt signaling pathway.

Pain is one of the common symptoms in patients diagnosed with MIRI, especially among elderly individuals. The physiological reaction to pain is the activation of the sympathetic nervous system. The enhanced sympathoexcitation results in increased cardiovascular workload, oxygen demand, and other detrimental effects which in turn aggravate the pain. During myocardial ischemia and reperfusion periods, multiple sources of algogenic substances, such as hydroxyl radicals, serotonin, hydrogen ions, potassium ions and cytokines, lead to activation of sympathetic nerve endings [42]. Based on the results of behavioral testing and serum NE measurement results in the study, AC improved the threshold of mechanical pain, which may be due to decreasing sympathetic nerve activity. As previously mentioned, AC exhibits anti-inflammation and antioxidant properties which are partly associated with amelioration of pain threshold and enhancing the tolerance to MIRI.

Collectively, the results of the present study indicated that AC pre-administration ameliorated pain and pathological changes in myocardial structure and decreased the serum levels of CK-MB, LDH and cTnl, as well as NE in MIRI rats. AC activated a variety of protective mechanisms against MIRI including suppression of apoptosis and oxidative stress, as well as regulation of the PI3K/Akt pathway (Fig. 7). As an initial exploration of AC’s cardioprotective effects against MIRI, this study has limitations including the mechanisms were confined to oxidative stress, apoptosis, and the PI3K/Akt pathway activation, the lack of in vitro validation, and the absence of a positive control. Future work will focus on identifying the novel pathways, the downstream of the PI3K/Akt pathway, and the interaction among key potential targets. In addition, the absence of a positive control weakens comparative validity and vitro experiment, which will be addressed in future studies.

Fig. 7
figure 7

A schematic representation outlines the potential impacts of AC on MIRI

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Data availability

Data is provided within the manuscript or supplementary information files.

Abbreviations

AC:

Acteoside

ACE:

Angiotensin converting enzyme

CK-MB:

Creatine kinase isoenzyme

cTnl:

Cardiac troponin l

GO:

Gene Ontology (GO)

HE:

Hematoxylin and eosin

KEGG:

Kyoto encyclopedia of genes and genomes

LAD:

Left anterior descending

LDH:

Lactate dehydrogenase

MDA:

Malondialdehyde

MI:

Myocardial infarction

MIRI:

Myocardial ischemia reperfusion injury

mTOR:

Mammalian target of rapamycin

NE:

Norepinephrine

NRF-2:

Nuclear factor signaling erythroid 2-related factor 2

PPI:

Protein-protein interaction

PWT:

Paw withdrawal threshold

SOD:

Superoxide dismutase

TUNEL:

Transferase-mediated dUTP nick end labeling

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Acknowledgements

Not applicable.

Funding

This study was supported by Natural Science Foundation of Heilongjiang Province (No. LH2022H091); Team Project of Fundamentals and Clinical Application about Cardiovascular Disease in the First Affiliated Hospital of Jiamusi University (No. 202301); Dongji academic team of Jiamusi University (DJXSTD202409); Open Project of Key Laboratory of Myocardial Ischemia, Ministry of Education (KF202116).

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Author notes

  1. Jing Li and Yuxin Guo contributed equally to this study.

Authors and Affiliations

  1. Department of Physiology, Basic medical college, Jiamusi University, 148 Xuefu Street, Jiamusi, Heilongjiang Province, 154000, P. R. China

    Jing Li, Yuxin Guo, Yang Yang & Lin-Lin Jia

  2. Department of Cardiology, the First affiliated hospital to Jiamusi University, 348 Dexiang Street Jiamusi, Jiamusi, Heilongjiang Province, 154000, P. R. China

    Qing Xue, Hong Cao, Guangyuan Yang & Hai-Bo Yu

  3. Zhangzhou Health Vocational College, No.29 Xiyangping Road, Xiangcheng District, Zhangzhou, Fujian Province, 363000, P. R. China

    Lin-Lin Jia

  4. Daqing Oilfield General Hospital, No. 195, Zhongqiao Road, Saltu District, Daqing City, Heilongjiang Province, 163000, P. R. China

    Zhiqi Sun

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  1. Jing Li
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Contributions

LLJ, HBY and GYY contributed towards the experimental design; JL, YXG, YY, QX and HC performed the experiments; HC JL and YXG analyzed the data; and JL, JJL, YXG, YY, QX, HC, HBY, GYY and ZQS wrote the manuscript. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Lin-Lin Jia or Hai-Bo Yu.

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Ethics approval and consent to participate

The experimental procedures for the care and use of laboratory animals were approved by the Biology and Animal Ethical Committee of the Basic Medical College of Jiamusi University (Jiamusi, China; approval no. 20240021). The procedures were conducted in accordance with the Guide for the Care and Use of Laboratory Animals from the National Institutes of Health.

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Li, J., Guo, Y., Yang, Y. et al. The cardioprotective effects of acteoside in myocardial ischemia reperfusion injury and the underlying mechanism. BMC Cardiovasc Disord 25, 329 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12872-025-04762-0

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Keywords

BMC Cardiovascular Disorders

ISSN: 1471-2261

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