Korean J Physiol Pharmacol.  2021 Mar;25(2):147-157. 10.4196/kjpp.2021.25.2.147.

Puerarin pretreatment attenuates cardiomyocyte apoptosis induced by coronary microembolization in rats by activating the PI3K/Akt/GSK-3β signaling pathway

• Chen ZQ
• Zhou Y
• Huang JW
• Chen F
• Zheng J
• Li HL
• Li T
• Li L

• KMID: 2512958
• DOI: http://doi.org/10.4196/kjpp.2021.25.2.147

Abstract

Coronary microembolization (CME) is associated with cardiomyocyte apoptosis and cardiac dysfunction. Puerarin confers protection against multiple cardiovascular diseases, but its effects and specific mechanisms on CME are not fully known. Hence, our study investigated whether puerarin pretreatment could alleviate cardiomyocyte apoptosis and improve cardiac function following CME. The molecular mechanism associated was also explored. A total of 48 Sprague-Dawley rats were randomly divided into CME, CME + Puerarin (CME + Pue), sham, and sham + Puerarin (sham + Pue) groups (with 12 rats per group). A CME model was established in CME and CME + Pue groups by injecting 42 μm microspheres into the left ventricle of rats. Rats in the CME + Pue and sham + Pue groups were intraperitoneally injected with puerarin at 120 mg/kg daily for 7 days before operation. Cardiac function, myocardial histopathology, and cardiomyocyte apoptosis index were determined via cardiac ultrasound, hematoxylin-eosin (H&E) and hematoxylin-basic fuchsin-picric acid (HBFP) stainings, and TdT-mediated dUTP nick-end labeling (TUNEL) staining, respectively. Western blotting was used to measure protein expression related to the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway. We found that, puerarin significantly ameliorated cardiac dysfunction after CME, attenuated myocardial infarct size, and reduced myocardial apoptotic index. Besides, puerarin inhibited cardiomyocyte apoptosis, as revealed by decreased Bax and cleaved caspase-3, and up-regulated Bcl-2 and PI3K/Akt/GSK-3β pathway related proteins. Collectively, puerarin can inhibit cardiomyocyte apoptosis and thus attenuate myocardial injury caused by CME. Mechanistically, these effects may be achieved through activation of the PI3K/Akt/GSK-3β pathway.

Keyword

Apoptosis; Coronary microembolization; Myocardial injury; PI3K/Akt/GSK-3; Puerarin

Figure

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  Fig. 1 The chemical structure of puerarin.

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  Fig. 2 Echocardiographic analysis of rats in four groups. (A–D) Sham, sham + Pue, CME, CME + Pue group, respectively. Left ventricular fractional shortening (LVFS), left ventricular end-diastolic diameter (LVEDd), left ventricular ejection fraction (LVEF), and left ventricular end-systolic diameter (LVESd) were quantitatively analyzed. CME, coronary microembolization; Pue, puerarin. ap < 0.05 in comparison with the sham group; bp < 0.05 in comparison with the CME group.

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  Fig. 3 Myocardial infarct size as shown by HBFP staining. (A–D) Sham, sham + Pue, CME, CME + Pue group, respectively. Ischemic myocardium was stained red. The arrows in (C and D) indicate the microinfarct size. CME, coronary microembolization; Pue, puerarin. ap < 0.05 in comparison with the sham group; bp < 0.05 in comparison with the CME group.

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  Fig. 4 Histopathological examination of myocardial tissue by H&E staining. (A–D) Sham, sham + Pue, CME, CME + Pue group, respectively. Microspheres with inflammatory cells infiltration were detected in both the CME and CME + Pue groups, but these pathological findings were not observed in the sham and sham + Pue groups. The arrows in (C and D) indicate microspheres. CME, coronary microembolization; Pue, puerarin.

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  Fig. 5 TUNEL staining of myocardial apoptosis in the four groups. (A–D) Sham, sham + Pue, CME, CME + Pue group, respectively. Data are expressed as mean ± standard deviation. Nuclei of apoptotic cardiomyocytes were yellow-brown, whereas the normal cardiomyocytes that did not show apoptosis were light blue. The arrows in (C and D) indicate the nuclei of apoptotic cardiomyocytes. CME, coronary microembolization; Pue, puerarin. ap < 0.05 in comparison with the sham group; bp < 0.05 in comparison with the CME group.

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  Fig. 6 Puerarin reduced cardiomyocyte apoptosis by regulating cleaved caspase-3, Bax as well as Bcl-2 after CME. Values are presented as mean ± standard deviation. The data were determined by at least three independent experiments in the present study. CME, coronary microembolization; Pue, puerarin. ap < 0.05 in comparison with the sham group; bp < 0.05 in comparison with the CME group.

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  Fig. 7 Effects of puerarin on PI3K/Akt/GSK-3β pathway. Values are expressed as mean ± standard deviation. The data were determined by at least three independent experiments in the present study. CME, coronary microembolization; Pue, puerarin. ap < 0.05 in comparison with the sham group; bp < 0.05 in comparison with the CME group.

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  Fig. 8 The graphical abstract shows that puerarin inhibits cardiomyocyte apoptosis induced by coronary microembolization in rats by activating the PI3K/Akt/GSK-3β signaling pathway.

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