Hesperetin alleviates high glucose and high lipid induced mitochondrial damage and apoptosis in AC16 cardiomyocytes via the AMPKα/SIRT1/PGC‐1α signaling pathway

XU Feixue; ZHANG Meng; LI Wanyi; GUO Yingying; XU Man; TANG Qizhu

doi:10.14188/j.1671-8852.2025.0781

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Hesperetin alleviates high glucose and high lipid induced mitochondrial damage and apoptosis in AC16 cardiomyocytes via the AMPKα/SIRT1/PGC‐1α signaling pathway

更新时间：2026-11-05

- Hesperetin alleviates high glucose and high lipid induced mitochondrial damage and apoptosis in AC16 cardiomyocytes via the AMPKα/SIRT1/PGC‐1α signaling pathway
- Medical Journal of Wuhan University Vol. 47, Issue 10, Pages: 1302-1309(2026)
- 作者机构：
  
  武汉大学人民医院心血管内科/代谢与相关慢病湖北省重点实验室湖北武汉 430060
- 作者简介：
- 基金信息：
- DOI：10.14188/j.1671-8852.2025.0781
  CLC： R54
- Received：15 July 2025，
  
  Published：25 October 2026
- 稿件说明：
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徐飞雪,张梦,李婉怡等.橙皮素通过AMPKα/SIRT1/PGC⁃1α通路改善高糖高脂诱导的AC16心肌细胞线粒体损伤与凋亡[J].武汉大学学报（医学版）,2026,47(10):1302-1309.

XU Feixue,ZHANG Meng,LI Wanyi,et al.Hesperetin alleviates high glucose and high lipid induced mitochondrial damage and apoptosis in AC16 cardiomyocytes via the AMPKα/SIRT1/PGC‐1α signaling pathway[J].Medical Journal of Wuhan University,2026,47(10):1302-1309.
徐飞雪,张梦,李婉怡等.橙皮素通过AMPKα/SIRT1/PGC⁃1α通路改善高糖高脂诱导的AC16心肌细胞线粒体损伤与凋亡[J].武汉大学学报（医学版）,2026,47(10):1302-1309. DOI： 10.14188/j.1671-8852.2025.0781.

XU Feixue,ZHANG Meng,LI Wanyi,et al.Hesperetin alleviates high glucose and high lipid induced mitochondrial damage and apoptosis in AC16 cardiomyocytes via the AMPKα/SIRT1/PGC‐1α signaling pathway[J].Medical Journal of Wuhan University,2026,47(10):1302-1309. DOI： 10.14188/j.1671-8852.2025.0781.

摘要

目的

研究橙皮素（HES）在高糖高脂诱导AC16心肌细胞氧化损伤中的影响及机制。

方法

体外培养AC16心肌细胞，使用不同浓度橙皮素处理细胞24 h后通过CCK‐8检测橙皮素对细胞活性的影响。使用300 μmol·L

-1

棕榈酸与4.5 g·L

-1

‐葡萄糖构建AC16细胞损伤模型。不同浓度橙皮素干预高糖高脂损伤细胞24 h后通过CCK‐8试验及ELISA法检测细胞上清液乳酸脱氢酶(LDH)水平选取有效的干预浓度。随后细胞随机分为5组：对照组、橙皮素组、高糖高脂组、高糖高脂+橙皮素组、高糖高脂+橙皮素+AMPK抑制剂Compound C（CC）组。通过DHE染色法检测细胞活性氧（ROS）蓄积，TUNEL染色法检测细胞凋亡率，JC‐1荧光探针染色法检测细胞线粒体膜电位，MitoSOX RED荧光探针染色法检测线粒体氧化应激，ELISA法检测细胞上清液丙二醛（MDA）水平和超氧化物歧化酶（SOD）活性，ATP试剂盒检测细胞ATP水平，Western Blot检测B细胞淋巴瘤‐2（Bcl‐2）、Bcl‐2相关X蛋白（Bax）、

AMPK、p‐AMPK、沉默信息调节因子1(SIRT1)、过氧化物酶体增殖物激活受体γ共激活因子‐1α(PGC‐1α)、解偶联蛋白3（UCP3）蛋白表达变化。

结果

和对照组比较，高糖高脂组心肌细胞活性显著降低，且心肌损伤标志物水平显著升高(

0.05)；细胞凋亡增加，Bax水平升高而Bcl‐2水平降低，活性氧增多、线粒体膜电位降低、线粒体氧化应激升高、MDA水平升高、SOD活性降低、ATP水平降低(

0.05)，AMPKα磷酸化水平、SIRT1、PGC‐1α、UCP3表达降低(

0.05)。与高糖高脂组比较，高糖高脂+橙皮素组心肌细胞活性明显增加，且心肌损伤标志物水平降低(

0.05)；细胞凋亡减少，Bax水平降低而Bcl‐2水平升高，活性氧减少、线粒体膜电位升高、线粒体氧化应激降低、MDA水平降低、SOD活性升高、ATP水平升高(

0.05)，且AMPKα磷酸化水平、SIRT1、PGC‐1α、UCP3表达升高(

0.05)，而使用AMPK抑制剂Compound C能削弱HES对上述通路蛋白的激活作用(

0.05)。

结论

橙皮素能够改善高糖高脂诱导的AC16心肌细胞线粒体损伤、氧化应激及细胞凋亡，其作用机制可能与激活AMPKα/SIRT1/PGC‐1α/UCP3信号通路有关。

Abstract

Objective

To investigate the effect and mechanism of hesperetin (HES) on oxidative damage induced by high glucose and high fat in AC16 cardiomyocytes.

Methods

AC16 cardiomyocytes were cultured

in vitro

and treated with different concentrations of hesperetin for 24 h. The effect of hesperetin on cell viability was detected by CCK⁃8. A cellular damage model was established using 300 μmol·L

¹ palmitic acid and 4.5 g·L

⁃glucose. After treating high glucose⁃ and high fat⁃damaged cells with different concentrations of hesperetin for 24 h

effective concentrations were selected based on CCK⁃8 assay and ELISA measurement of lactate dehydrogenase (LDH) levels in the cell supernatant. Subsequently

cells were randomly divided into 5 groups: control group

hesperetin alone group

high glucose and high fat group

high glucose and high fat+hesperetin group

and high glucose and high fat+hesperetin+AMPK inhibitor Compound C (CC) group. Reactive oxygen species (ROS) accumulation was detected by DHE staining

apoptosis rate was detected by TUNEL staining

mitochondrial membrane potential was detected

by JC⁃1 fluorescent probe staining

mitochondrial oxidative stress was detected by MitoSOX Red fluorescent probe staining

malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity in the cell supernatant were detected by ELISA

cellular ATP levels were detected by an ATP assay kit

and protein expression of B⁃cell lymphoma 2 (Bcl⁃2)

Bcl⁃2⁃associated X protein (Bax)

AMPK

p⁃AMPK

silent information regulator 1 （SIRT1)

peroxisome proliferator⁃activated receptor gamma coactivator 1⁃alpha (PGC⁃1α)

and uncoupling protein 3 (UCP3） was detected by Western Blot.

Results

Compared with the control group

the high glucose and high fat group showed significantly reduced cardiomyocyte viability and increased levels of myocardial injury markers (

0.05)

as well as increased apoptosis

increased Bax but decreased Bcl⁃2 levels

increased reactive oxygen species

decreased mitochondrial membrane potential

increased mitochondrial oxidative stress

increased MDA levels

decreased SOD activity

decreased ATP levels (

0.05)

and decreased phosphorylation of AMPKα and expression of SIRT1

PGC⁃1α

and UCP3 (

0.05). Compared with the high glucose and high fat group

the high glucose and high fat+hesperetin group showed significantly increased cardiomyocyte viability and decreased levels of myocardial injury markers (

0.05)

as well as decreased apoptosis

decreased Bax but increased Bcl⁃2 levels

decreased reactive oxygen species

increased mitochondrial membrane potential

decreased mitochondrial oxidative stress

decreased MDA levels

increased SOD activity

increased ATP levels (

0.05)

and increased phosphorylation of AMPKα and expression of SIRT1

PGC⁃1α

and UCP3 (

0.05). Furthermore

the use of the AMPK inhibitor Compound C attenuated the activating effect of HES on the above pathway proteins (

0.05).

Conclusion

Hesperetin can ameliorate mitochondrial damage

oxidative stress

and apoptosis induced by high glucose and high fat in AC16 cardiomyocytes

and its mechanism may involve the activation of the AMPKα/SIRT1/PGC⁃1α/UCP3 signaling pathway.

关键词

Keywords

references

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