Isovitexin alleviates myocardial oxidative stress injury in diabetic mice by enhancing myocardial SIRT3 expression and reducing oxidative stress

doi:10.12122/j.issn.1673-4254.2026.02.09

Abstract

Abstract:

Objective To investigate the protective effect of isovitexin (ISO) on the myocardium of diabetic mice and explore its mechanism. Methods Eighteen adult male C57 mice were randomly divided into control group, diabetes mellitus (DM) group and DM+ISO group (n=6). Primary cultures of neonatal mouse cardiomyocytes (NMCMs) were exposed to high glucose (33 mmol/L) and treated with ISO alone or in combination with 3-TYP (a SIRT3 inhibitor). HE staining was used to evaluate myocardial injury in the diabetic mice, and the levels of MDA, SOD and GSH-Px in the myocardium were detected with ELISA. The expressions of iNOS, NOX2 and 8-OHdG were detected using immunohistochemistry and fluorescence staining. Western blotting was used to analyze the expression levels of NRF2, NOX2, NQO1, and SIRT3, and ROS levels were determined with flow cytometry. Results The diabetic mice showed obvious inflammatory cell infiltration in the myocardium, increased myocardial levels of IL-1β, IL-6 and TNF‑α, decreased expression levels of NQO1, NRF2 and SIRT3 proteins, and increased expression levels of NOX2 and AC-SOD2 proteins. ISO treatment of the diabetic mice significantly reduced myocardial inflammatory cell infiltration, lowered the levels of IL-1β, IL-6 and TNF‑α, restored the protein levels of NQO1, NRF2 and SIRT3, and decreased the protein levels of NOX2 and AC-SOD2. Conclusion ISO can alleviate diabetic myocardial injury in mice by promoting SIRT3 expression and reducing oxidative stress.

Key words: isovitexin, diabetes mellitus, cardiomyopathy, oxidative stress, SIRT3

Yuce PENG, Yi JIANG, Dan MA, An HE, Dingyi LÜ, Minghao LUO, Suxin LUO. Isovitexin alleviates myocardial oxidative stress injury in diabetic mice by enhancing myocardial SIRT3 expression and reducing oxidative stress[J]. Journal of Southern Medical University, 2026, 46(2): 316-324.

Figures/Tables 5

Fig.1 Isovitexin (ISO) alleviates myocardial injury in diabetic mice. A: HE staining of the myocardial tissues of mice (Scale bar=50 μm). B: MDA content in mouse myocardial tissue (n=6). C: GSH-Px content in mouse myocardial tissue (n=6). D: SOD content in mouse myocardial tissue (n=6). E, F: Expression levels of NQO1, NRF2, AC-SOD2, SIRT3, and NOX2 proteins detected by Western blotting (n=4). DM: Diabetes mellitus. ***P<0.001 vs Sham group; #P<0.05, ##P<0.01, ###P<0.001 vs DM group.

Fig.2 ISO alleviates myocardial oxidative stress in diabetic mice. A: Expression and location of 8-OHdG determined by immunofluorescence staining (Scale bar=100 μm). B: Statistical analysis of fluorescence intensity (n=6). C: Expression and location of iNOS determined by immunohistochemistry (Scale bar=100 μm). D: Statistical analysis of iNOS-positive area (n=6). E: Expression and location of NOX4 determined by immunohistochemistry (Scale bar=100 μm). F: Statistical analysis of NOX4-positive area (n=6). ***P<0.001 vs Sham group; ##P<0.01, ###P<0.001 vs DM group.

Fig.3 ISO alleviates oxidative stress in primary cultures of neonatal mouse cardiomyocytes (NMCMs). A: Intracellular ROS levels determined by immunofluorescence staining (Scale bar=50 μm). B: Statistical analysis of fluorescence intensity (n=6). C, D: Expression levels of SIRT3, NQO1, AC-SOD2, NRF2, and NOX2 proteins detected by Western blotting (n=6). ***P<0.001 vs CON group; ##P<0.01, ###P<0.001 vs HG group.

Fig.4 ISO protects NMCMs against high glucose-induced injury by activating SIRT3. A, B: Expression levels of SIRT3, NQO1, AC-SOD2, NRF2, and NOX2 detected by Western blotting (n=6). C, D: Flow cytometry for quantitative analysis of cellular oxidative stress damage (n=6). ***P<0.001 vs CON group; ##P<0.01, ###P<0.001 vs HG group; &P<0.05, &&P<0.01 vs HG+ISO group.

Fig5 ISO protects myocardial tissue of diabetic mice against oxidative stress by activating SIRT3. A: MDA content in mouse myocardial tissues (n=6). B: GSH-Px content in mouse myocardial tissues (n=6). C: SOD content in mouse myocardial tissue (n=6). D: Expression levels of NQO1, NRF2, AC-SOD2, SIRT3, and NOX2 proteins in mouse myocardial tissues detected using Western blotting (n=6). Comparisons were performed using one-way ANOVA for multiple groups. ##P<0.01, ###P<0.001 vs DM group; &&P<0.01, &&&P<0.001 vs DM+ISO group.

References 31

[1]	Saeedi P, Petersohn I, Salpea P, et al. Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas, 9(th) edition [J]. Diabetes Res Clin Pract, 2019, 157: 107843. doi：10.1016/j.diabres.2019.107843
[2]	Qin FZ, Siwik DA, Luptak I, et al. The polyphenols resveratrol and S17834 prevent the structural and functional sequelae of diet-induced metabolic heart disease in mice[J]. Circulation, 2012, 125(14): 1757-64, S1-6. doi：10.1161/circulationaha.111.067801
[3]	Kang QZ, Yang CX. Oxidative stress and diabetic retinopathy: molecular mechanisms, pathogenetic role and therapeutic implications[J]. Redox Biol, 2020, 37: 101799. doi：10.1016/j.redox.2020.101799
[4]	Zhang GY, Wang XD, Li C, et al. Integrated stress response couples mitochondrial protein translation with oxidative stress control[J]. Circulation, 2021, 144(18): 1500-15. doi：10.1161/circulationaha.120.053125
[5]	Heinke L. Mitochondrial ROS drive cell cycle progression[J]. Nat Rev Mol Cell Biol, 2022, 23(9): 581. doi：10.1038/s41580-022-00523-5
[6]	Jin LG, Geng LL, Ying L, et al. FGF21-sirtuin 3 axis confers the protective effects of exercise against diabetic cardiomyopathy by governing mitochondrial integrity[J]. Circulation, 2022, 146(20): 1537-57. doi：10.1161/circulationaha.122.059631
[7]	Lv HM, Yu ZX, Zheng YW, et al. Isovitexin exerts anti-inflammatory and anti-oxidant activities on lipopolysaccharide-induced acute lung injury by inhibiting MAPK and NF‑κB and activating HO-1/Nrf2 pathways[J]. Int J Biol Sci, 2016, 12(1): 72-86. doi：10.7150/ijbs.13188
[8]	Lin JQ, Ran H, Feng QQ, et al. Unveiling the differences between vitexin and isovitexin: from the perspective of sources, green advanced extraction technologies, biological activities, and safety[J]. Food Chem, 2025, 485: 144600. doi：10.1016/j.foodchem.2025.144600
[9]	He M, Min JW, Kong WL, et al. A review on the pharmacological effects of vitexin and isovitexin[J]. Fitoterapia, 2016, 115: 74-85. doi：10.1016/j.fitote.2016.09.011
[10]	Peng F, Liao MR, Jin WK, et al. 2-APQC, a small-molecule activator of Sirtuin-3 (SIRT3), alleviates myocardial hypertrophy and fibrosis by regulating mitochondrial homeostasis[J]. Signal Transduct Target Ther, 2024, 9(1): 133. doi：10.1038/s41392-024-01816-1
[11]	Zhang J, Ye J, Zhu SO, et al. Context-dependent role of SIRT3 in cancer[J]. Trends Pharmacol Sci, 2024, 45(2): 173-90. doi：10.1016/j.tips.2023.12.005
[12]	Palomer X, Román-Azcona MS, Pizarro-Delgado J, et al. SIRT3-mediated inhibition of FOS through histone H3 deacetylation prevents cardiac fibrosis and inflammation[J]. Signal Transduct Target Ther, 2020, 5(1): 14. doi：10.1038/s41392-020-0114-1
[13]	Gao P, You M, Li L, et al. Salt-induced hepatic inflammatory memory contributes to cardiovascular damage through epigenetic modulation of SIRT3[J]. Circulation, 2022, 145(5): 375-91. doi：10.1161/circulationaha.121.055600
[14]	Jheng JR, Bai Y, Noda K, et al. Skeletal muscle SIRT3 deficiency contributes to pulmonary vascular remodeling in pulmonary hypertension due to heart failure with preserved ejection fraction[J]. Circulation, 2024, 150(11): 867-83. doi：10.1161/circulationaha.124.068624
[15]	Xie SY, Liu SQ, Zhang T, et al. USP28 serves as a key suppressor of mitochondrial morphofunctional defects and cardiac dysfunction in the diabetic heart[J]. Circulation, 2024, 149(9): 684-706. doi：10.1161/circulationaha.123.065603
[16]	Fan YN, Chen ZW, Wang HX, et al. Isovitexin targets SIRT3 to prevent steroid-induced osteonecrosis of the femoral head by modulating mitophagy-mediated ferroptosis[J]. Bone Res, 2025, 13(1): 18. doi：10.1038/s41413-024-00390-0
[17]	Peng YC, Guo MY, Luo MH, et al. Dapagliflozin ameliorates myocardial infarction injury through AMPKα-dependent regulation of oxidative stress and apoptosis[J]. Heliyon, 2024, 10(7): e29160. doi：10.1016/j.heliyon.2024.e29160
[18]	Rubler S, Dlugash J, Yuceoglu YZ, et al. New type of cardiomyopathy associated with diabetic glomerulosclerosis[J]. Am J Cardiol, 1972, 30(6): 595-602. doi：10.1016/0002-9149(72)90595-4
[19]	Kautzky-Willer A, Leutner M, Harreiter J. Sex differences in type 2 diabetes[J]. Diabetologia, 2023, 66(6): 986-1002. doi：10.1007/s00125-023-05891-x
[20]	Tan Y, Zhang ZG, Zheng C, et al. Mechanisms of diabetic cardiomyopathy and potential therapeutic strategies: preclinical and clinical evidence[J]. Nat Rev Cardiol, 2020, 17(9): 585-607. doi：10.1038/s41569-020-0339-2
[21]	Wang DZ, Yin Y, Wang SY, et al. FGF1^ΔHBS prevents diabetic cardiomyopathy by maintaining mitochondrial homeostasis and reducing oxidative stress via AMPK/Nur77 suppression[J]. Signal Transduct Target Ther, 2021, 6(1): 133. doi：10.1038/s41392-021-00542-2
[22]	Kyriazis ID, Hoffman M, Gaignebet L, et al. KLF5 is induced by FOXO1 and causes oxidative stress and diabetic cardiomyopathy[J]. Circ Res, 2021, 128(3): 335-57. doi：10.1161/circresaha.120.316738
[23]	Wang MY, Zhang SW, Tian JW, et al. Impaired iron-sulfur cluster synthesis induces mitochondrial PARthanatos in diabetic cardiomyopathy[J]. Adv Sci (Weinh), 2025, 12(1): e2406695. doi：10.1002/advs.202406695
[24]	Huang YS, Luo W, Chen SY, et al. Isovitexin alleviates hepatic fibrosis by regulating miR-21-mediated PI3K/Akt signaling and glutathione metabolic pathway: based on transcriptomics and metabolomics[J]. Phytomedicine, 2023, 121: 155117. doi：10.1016/j.phymed.2023.155117
[25]	Hao RL, Li MQ, Li F, et al. Protective effects of the phenolic compounds from mung bean hull against H₂O₂-induced skin aging through alleviating oxidative injury and autophagy in HaCaT cells and HSF cells[J]. Sci Total Environ, 2022, 841: 156669. doi：10.1016/j.scitotenv.2022.156669
[26]	Pal S, Singh M, Porwal K, et al. Adiponectin receptors by increasing mitochondrial biogenesis and respiration promote osteoblast differentiation: Discovery of isovitexin as a new class of small molecule adiponectin receptor modulator with potential osteoanabolic function[J]. Eur J Pharmacol, 2021, 913: 174634. doi：10.1016/j.ejphar.2021.174634
[27]	Zhu J, Sun RP, Sun KQ, et al. The deubiquitinase USP11 ameliorates intervertebral disc degeneration by regulating oxidative stress-induced ferroptosis via deubiquitinating and stabilizing Sirt3[J]. Redox Biol, 2023, 62: 102707. doi：10.1016/j.redox.2023.102707
[28]	Ning Y, Dou XY, Wang ZC, et al. SIRT3: a potential therapeutic target for liver fibrosis[J]. Pharmacol Ther, 2024, 257: 108639. doi：10.1016/j.pharmthera.2024.108639
[29]	D'Onofrio N, Prattichizzo F, Marfella R, et al. SIRT3 mediates the effects of PCSK9 inhibitors on inflammation, autophagy, and oxidative stress in endothelial cells[J]. Theranostics, 2023, 13(2): 531-42. doi：10.7150/thno.80289
[30]	Mihanfar A, Nejabati HR, Fattahi A, et al. SIRT3-mediated cardiac remodeling/repair following myocardial infarction[J]. Biomed Pharmacother, 2018, 108: 367-73. doi：10.1016/j.biopha.2018.09.079
[31]	Su H, Cantrell AC, Chen JX, et al. SIRT3 deficiency enhances ferroptosis and promotes cardiac fibrosis via p53 acetylation[J]. Cells, 2023, 12(10): 1428. doi：10.3390/cells12101428