异牡荆素通过促进SIRT3表达减轻糖尿病小鼠的心肌氧化应激损伤

doi:10.12122/j.issn.1673-4254.2026.02.09

摘要/Abstract

摘要：

目的研究异牡荆素（ISO）对糖尿病小鼠心肌的保护作用及机制。方法选用成年雄性C57小鼠18只，随机分为对照组、糖尿病组（DM）和DM+ISO组（n=6）。分离新生小鼠原代心肌细胞（NMCMs），分为对照组、高糖组（HG，33 mmol/L葡萄糖）、HG+ISO组及HG+ISO+3-TYP组（SIRT3抑制剂）。HE染色评估心肌损伤；ELISA检测心肌丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）水平；免疫组化和荧光检测iNOS、NOX2、8-OHdG表达；Western blotting分析NRF2、NOX2、NQO1、SIRT3等通路蛋白；流式细胞术定量活性氧水平。结果与对照组相比，DM组心肌细胞炎症浸润增加，白细胞介素-1β（IL-1β），白细胞介素-6（IL-6），α-肿瘤坏死因子（TNF-α）水平升高（P<0.01），NQO1，NRF2，SIRT3蛋白水平下降（P<0.01），NOX2，AC-SOD2蛋白水平增加（P<0.01），与DM组相比，DM+ISO组心肌细胞炎症浸润减少，IL-1β，IL-6，TNF-α水平下降（P<0.01），NQO1，NRF2，SIRT3蛋白水平恢复（P<0.01），NOX2，AC-SOD2蛋白水平减少（P<0.01）。结论 ISO可通过促进SIRT3表达，有效减轻氧化应激来减缓糖尿病心肌损伤，从而显著改善糖尿病心肌病。

关键词: 异牡荆素, 糖尿病, 心肌病, 氧化应激, SIRT3

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

彭煜策, 姜毅, 马丹, 何岸, 吕鼎一, 罗明昊, 罗素新. 异牡荆素通过促进SIRT3表达减轻糖尿病小鼠的心肌氧化应激损伤[J]. 南方医科大学学报, 2026, 46(2): 316-324.

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.

图/表 5

图1 ISO减轻糖尿病小鼠心肌损伤

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.

图2 ISO减轻糖尿病小鼠心肌氧化应激

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.

图3 ISO减轻新生小鼠心肌细胞氧化应激

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.

图4 ISO通过激活SIRT3在体外发挥保护作用

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.

图5 ISO通过激活SIRT3在体内发挥保护作用

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.

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