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Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway

  • Authors:
    • Xiaoyu Zheng
    • Jinping Li
    • Tianyang Hu
    • Li Tan
    • Ding Lan
    • Ying Deng
  • View Affiliations / Copyright

    Affiliations: Department of Cardiovascular and Cerebrovascular Diseases, School of Clinical Medicine, Chongqing Medical and Pharmaceutical College, Chongqing 401331, P.R. China, Department of Science and Education, The First People's Hospital of Zhaotong City and Zhaotong Hospital Affiliated to Kunming Medical University, Zhaotong, Yunnan 657000, P.R. China, Department of Precision Medicine Center, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China, Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China
    Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 114
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    Published online on: February 11, 2026
       https://doi.org/10.3892/mmr.2026.13824
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Abstract

Myocardial ischemia‑reperfusion injury (MIRI) can trigger inflammatory responses and cause pyroptosis. Puerarin (Pue), as a traditional medicine, exhibits potential value in cardiac protection. However, the mechanism by which Pue regulates pyroptosis in MIRI remains to be fully elucidated. The present study aimed to explore the cardioprotective effects of Pue against MIRI and reveal the underlying mechanisms of these effects. Sprague‑Dawley rats were used to establish in vivo models of MIRI, while H9C2 rat embryonic cardiomyocytes were employed as in vitro models. Echocardiography was performed to measure cardiac function. Triphenyltetrazolium chloride/Evans blue staining, hematoxylin‑eosin staining, Masson's trichrome staining and immunohistochemistry were employed to assess the pharmacodynamic effects of Pue. The expression of molecules related to pyroptosis, such as nuclear factor E2‑related factor 2 (NRF2) and heme oxygenase‑1 (HO‑1) were detected by immunofluorescence, Hoechst 33342/PI staining, reverse transcription‑quantitative PCR and western blot analyses. The results of the present study showed that Pue pretreatment reduced the area of myocardial infarction and decreased the expression of pyroptosis‑related molecules. Additionally, Pue was shown to reverse H2O2‑induced mitochondrial dysfunction in cardiomyocytes and inhibit nucleotide‑binding oligomerization domain‑like receptor family pyrin domain‑containing 3 (NLRP3)/caspase‑1/gasdermin D (GSDMD)‑mediated pyroptosis. Pue was also shown to stimulate the nuclear translocation of NRF2 and increase the expression of HO‑1. Furthermore, Pue further demonstrated its anti‑pyroptotic effects by activating the NRF2/HO‑1 pathway. The present study revealed that Pue can protect injured myocardium after MIRI by inhibiting NLRP3/caspase‑1/GSDMD‑mediated pyroptosis. The mechanism of action for these cardioprotective effects relied upon downregulation of the NRF2/HO‑1 signaling pathway. The findings of the present study provided a novel strategy for the clinical application of puerarin in the treatment of MIRI.
View Figures

Figure 1

Pue displays cardioprotective effects
against myocardial ischemia-reperfusion injury in vivo. (A)
Timeline of Pue administration and schematic of the rat I/R injury
model. (B) Changes in cardiac function (LVIDd, LVIDs, LVEF) and
heart rate were observed in each group at 2 h after myocardial I/R.
(C) Triphenyltetrazolium chloride staining of myocardial tissues.
(D) Quantitative comparison of infarction areas among each group
(n=8). (E) Masson's trichrome staining of myocardial tissues (scale
bar, 100 µm). **P<0.01, ***P<0.001 and ****P<0.0001 vs.
sham group; ##P<0.01, ###P<0.001 and
####P<0.0001 vs. I/R group. Pue, puerarin; LVIDd,
left ventricular internal dimension at end-diastole; LVIDs, left
ventricular internal dimension at end-systole; LVEF, left
ventricular ejection fraction; I/R, ischemia/reperfusion.

Figure 2

Pue inhibits the key proteins of the
inflammasome and pyroptosis in myocardial ischemia-reperfusion
injury. (A) Hematoxylin-eosin staining showed the inflammatory
condition of myocardial tissue (n=8; scale bar, 100 µm). (B)
Immunohistochemistry detected the positive expression of
inflammasome markers in myocardial tissues of each group (scale
bar, 100 µm). (C) The AOD values of inflammasome markers obtained
by immunohistochemical staining. (D) Expression of pyroptotic
proteins and HO-1. (E) Quantification of relative mRNA levels of
key pyroptotic markers. (F) Semi-quantification of relative
expression levels of pyroptotic proteins and HO-1. (G) Protein
expression of nuclear NRF2 was assessed by western blotting. (H)
Semi-quantification of relative expression levels of NRF2 obtained
by western blotting. **P<0.01, ***P<0.001 and ****P<0.0001
vs. sham group; ####P<0.0001 vs. I/R group.. AOD,
average optical density; HO-1, heme oxygenase-1; ASC,
apoptosis-associated speck-like protein; NLRP3, nucleotide-binding
oligomerization domain-like receptor family pyrin domain-containing
3; GSDMD, gasdermin D; Pue, puerarin; I/R,
ischemia/reperfusion.

Figure 3

Pue inhibits pyroptosis in
cardiomyocytes. (A) JC-1 was used to detect the mitochondrial
membrane potential of cardiomyocytes (scale bar, 100 µm). (B)
Quantitative analysis of the ratio of red to green fluorescence.
(C) Western blot analysis showed the expression levels of key
proteins involved in pyroptosis in cardiomyocytes. (D)
Representative morphology of mitochondria obtained by transmission
electron microscopy (scale bar, 1 µm). The green arrow showed
normal mitochondria, while the red arrow indicated organelle
vacuolation. (E) Semi-quantification of the relative expression
levels of key pyroptotic proteins in cardiomyocytes from western
blotting results. ****P<0.0001 vs. control group;
##P<0.01 and ####P<0.0001 vs.
H2O2 group. Pue, puerarin; ASC,
apoptosis-associated speck-like protein; GSDMD, gasdermin D; NLRP3,
nucleotide-binding oligomerization domain-like receptor family
pyrin domain-containing 3.

Figure 4

Pue activates the NRF2 pathway to
alleviate cardiomyocyte pyroptosis. (A) Western blot assays were
used to detect NRF2 expression in cardiomyocytes of the OE-NC group
and the OE-NRF2 group (n=3). (B) Semi-quantification of NRF2
protein levels detected in western blotting. (C) Immunofluorescence
was used to detect nuclear translocation of NRF2 in each group of
cells (n=3). (D) HO-1 expression was detected by immunofluorescence
staining in each group of cells (n=3). (E) Quantitative analysis of
the fluorescence intensities of NRF2 and HO-1 immunofluorescence
staining. (F) Activity of LDH in each group. **P<0.01 and
****P<0.0001 vs. OE-NC group; ###P<0.001 and
####P<0.0001 vs. H2O2 + OE-NC
group. OE, overexpression; NC, negative control; NRF2, nuclear
factor E2-related factor 2; Pue, puerarin; HO-1, heme oxygenase-1;
LDH, lactate dehydrogenase.

Figure 5

Pue alleviates cardiomyocyte
pyroptosis via the NRF2/HO-1 signaling pathway. (A) Hoechst
33342/PI staining detected the apoptotic state of H9C2 cells
(magnification, ×400) (n=3). (B) Quantitative analysis of
PI-positive cells. (C) Western blotting of nuclear NRF2 expression
in each group (n=3). (D) Western blotting images of key pyroptotic
proteins and HO-1 in each treatment group (n=3). (E)
Semi-quantification of the expression of pyroptotic proteins and
HO-1 in each group of cells. (F) Semi-quantification of NRF2
protein expression. ****P<0.0001 vs. OE-NC group;
###P<0.001 and ####P<0.0001 vs.
H2O2 + OE-NC group. Pue, puerarin; OE,
overexpression; NC, negative control; NRF2, nuclear factor
E2-related factor 2; ASC, apoptosis-associated speck-like protein;
NLRP3, nucleotide-binding oligomerization domain-like receptor
family pyrin domain-containing 3; GSDMD, gasdermin D; HO-1, heme
oxygenase-1.

Figure 6

Schematic diagram of the molecular
mechanism of Pue-mediated pyroptosis inhibition. NRF2, nuclear
factor E2-related factor 2; HO-1, heme oxygenase-1; ARE,
antioxidant response element; NLRP3, nucleotide-binding
oligomerization domain-like receptor family pyrin domain-containing
3; ASC, apoptosis-associated speck-like protein; GSDMD, gasdermin
D.
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Copy and paste a formatted citation
Spandidos Publications style
Zheng X, Li J, Hu T, Tan L, Lan D and Deng Y: Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway. Mol Med Rep 33: 114, 2026.
APA
Zheng, X., Li, J., Hu, T., Tan, L., Lan, D., & Deng, Y. (2026). Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway. Molecular Medicine Reports, 33, 114. https://doi.org/10.3892/mmr.2026.13824
MLA
Zheng, X., Li, J., Hu, T., Tan, L., Lan, D., Deng, Y."Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway". Molecular Medicine Reports 33.4 (2026): 114.
Chicago
Zheng, X., Li, J., Hu, T., Tan, L., Lan, D., Deng, Y."Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway". Molecular Medicine Reports 33, no. 4 (2026): 114. https://doi.org/10.3892/mmr.2026.13824
Copy and paste a formatted citation
x
Spandidos Publications style
Zheng X, Li J, Hu T, Tan L, Lan D and Deng Y: Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway. Mol Med Rep 33: 114, 2026.
APA
Zheng, X., Li, J., Hu, T., Tan, L., Lan, D., & Deng, Y. (2026). Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway. Molecular Medicine Reports, 33, 114. https://doi.org/10.3892/mmr.2026.13824
MLA
Zheng, X., Li, J., Hu, T., Tan, L., Lan, D., Deng, Y."Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway". Molecular Medicine Reports 33.4 (2026): 114.
Chicago
Zheng, X., Li, J., Hu, T., Tan, L., Lan, D., Deng, Y."Puerarin attenuates myocardial ischemia‑reperfusion injury by inhibiting myocardium pyroptosis via the NRF2/HO‑1 signaling pathway". Molecular Medicine Reports 33, no. 4 (2026): 114. https://doi.org/10.3892/mmr.2026.13824
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