Equol interferes with palmitic acid-induced muscle tube atrophy in L6 cells by regulating autophagy

Shengcai YU; Xiangmin NI; Heng LU; Guiming ZHANG; Jian WANG

doi:10.16016/j.2097-0927.202304058

您当前的位置：

首页 >

文章列表页 >

Equol interferes with palmitic acid-induced muscle tube atrophy in L6 cells by regulating autophagy

Monographic Reports: Nutritional intervention for chronic diseases | 更新时间：2026-01-09

- Equol interferes with palmitic acid-induced muscle tube atrophy in L6 cells by regulating autophagy
- Journal of Army Medical University Vol. 45, Issue 15, Pages: 1604-1611(2023)
- 作者机构：
  
  400037 重庆，陆军军医大学(第三军医大学)第二附属医院营养科
- 作者简介：
- 基金信息：
  
  the General Program of National Nature Science Foundation of China(81973040)
- DOI：10.16016/j.2097-0927.202304058
  CLC： R322.74;R592;R977.12
- Received：13 April 2023，
  
  Revised：2023-05-20，
  
  Published：15 August 2023
- 稿件说明：
移动端阅览
Shengcai YU, Xiangmin NI, Heng LU, et al. Equol interferes with palmitic acid-induced muscle tube atrophy in L6 cells by regulating autophagy[J]. Journal of Army Medical University, 2023, 45(15): 1604-1611.
DOI：

Shengcai YU, Xiangmin NI, Heng LU, et al. Equol interferes with palmitic acid-induced muscle tube atrophy in L6 cells by regulating autophagy[J]. Journal of Army Medical University, 2023, 45(15): 1604-1611. DOI： 10.16016/j.2097-0927.202304058.

摘要

目的

观察雌马酚(equol

Eq)干预对棕榈酸(palmitic acid，PA)诱导的L6大鼠成肌细胞肌管萎缩的影响，并探讨其机制。

方法

体外培养L6细胞，以不同浓度的Eq、PA处理24 h，通过CCK-8法检测相对细胞活力确定Eq、PA干预的适宜浓度。将细胞分为对照组、模型组(PA: 0.25 mmol/L)、干预组(PA: 0.25 mmol/L+Eq: 1 μmol/L)和自噬抑制剂组(PA: 0.25 mmol/L+Eq: 1 μmol/L+3-MA: 1 mmol/L)。干预24 h后检测各组细胞葡萄糖摄取能力；HE染色观察细胞肌管生长情况；透射电镜观察细胞自噬小体数量；激光共聚焦显微镜观察经线粒体荧光探针标记的线粒体形态；Western blot检测MyHC、p62蛋白、LC3蛋白、MFN2和DRP1表达水平。

结果

干预24 h后，与对照组比较，模型组L6细胞葡萄糖消耗明显减低(

＜0.05)，肌管直径明显减小(

＜0.05)，自噬小体数量减少，线粒体裂变增加，MyHC、MFN2蛋白表达水平降低(

＜0.05)，而p62、DRP1表达水平及LC3-Ⅰ/LC3-Ⅱ比值增加(

＜0.05)；与模型组相比，干预组L6细胞葡萄糖消耗明显增加(

＜0.05)，肌管直径明显增大(

＜0.05)，自噬小体数量增加，线粒体裂变减少，MyHC、MFN2蛋白表达水平升高(

＜0.05)，而p62、DRP1表达水平及LC3-Ⅰ/LC3-Ⅱ比值降低(

＜0.05)；Eq对PA诱导的L6细胞葡萄糖消耗，肌管直径，自噬小体数量，线粒体裂变，MyHC、p62、MFN2、DRP1蛋白表达水平及LC3-Ⅰ/LC3-Ⅱ比值的影响可被3-MA有效抑制。

结论

Eq可有效改善PA诱导的L6细胞肌管萎缩，其部分机制与Eq通过促进自噬，减轻线粒体裂变有关。

Abstract

Objective

To observe the effect of equol (Eq) intervention on palmitic acid (PA)-induced myoblast atrophy in rat myoblast cell line L6 and to explore its underlying mechanism.

Methods

L6 cells were divided into control group

model group (0.25 mmol/L PA)

intervention group (0.25 mmol/L PA+1 μmol/L Eq) and autophagy inhibitor (3-methyladenine

3-MA) group (0.25 mmol/L PA+1 μmol/L Eq+1 mmol/L 3-MA). After 24 h of intervention

glucose uptake capacity of cells was detected

muscle tube growth was observed by HE staining

autophagosome number was observed with transmission electron microscopy

and mitochondrial morphology labeled by fluorescent probe was observed by laser confocal microscopy. The expression levels of MyHC

p62

LC3

MFN2 and DRP1 were detected by Western blotting.

Results

Compared with the control group

24 h of PA treatment resulted in significantly decreased glucose consumption (

0.05)

decreased muscle tube diameter (

0.05)

reduced number of autophagosomes

increased mitochondrial fission

and reduced expression levels of MyHC and MFN2 (

0.05)

and elevated DRP1

p62 level and LC3-Ⅰ/LC3-Ⅱ ratio (

0.05). While Eq intervention reserved above changes induced by PA treatment (

0.05). However

the effects of Eq on PA-induced glucose consumption

myotube diameter

autophagosome number

mitochondrial fission

protein levels of MyHC

p62

MFN2

DRP1 and LC3-Ⅰ/LC3-Ⅱ ratio in L6 cells could be effectively inhibited by 3-MA treatment.

Conclusion

Eq effectively ameliorates PA-induced muscle tube atrophy in L6 cells

which may partially be related to Eq alleviating mitochondrial fission by promoting autophagy.

关键词

Keywords

references

FU J L, SUN L N, MU Z J, et al . Free fatty acids are associated with muscle dysfunction in Chinese adults with type 2 diabetes[J ] . Endocrine, 2022, 77(1): 41-47. DOI: 10.1007/s12020-022-03053-4.

IZZO A, MASSIMINO E, RICCARDI G, et al . A narrative review on sarcopenia in type 2 diabetes mellitus: prevalence and associated factors[J ] . Nutrients, 2021, 13(1): 183. DOI: 10.3390/nu13010183.

ANAGNOSTIS P, GKEKAS N K, ACHILLA C, et al . Type 2 diabetes mellitus is associated with increased risk of sarcopenia: a systematic review and meta-analysis[J ] . Calcif Tissue Int, 2020, 107(5): 453-463. DOI: 10.1007/s00223-020-00742-y.

AL SAEDI A, DEBRUIN D A, HAYES A, et al . Lipid metabolism in sarcopenia[J ] . Bone, 2022, 164: 116539. DOI: 10.1016/j.bone.2022.116539.

BELLANTI F, BUGLIO A L, VENDEMIALE G. Mitochondrial impairment in sarcopenia[J]. Biology, 2021, 10(1): 31. DOI: 10.3390/biology10010031.

CRUZ-JENTOFT A J, SAYER A A. Sarcopenia[J]. Lancet, 2019, 393(10191): 2636-2646. DOI: 10.1016/S0140-6736(19)31138-9.

HESSELINK M K C, SCHRAUWEN-HINDERLING V, SCHRAUWEN P. Skeletal muscle mitochondria as a target to prevent or treat type 2 diabetes mellitus[J]. Nat Rev Endocrinol, 2016, 12(11): 633-645. DOI: 10.1038/nrendo.2016.104.

NG M Y W, WAI T, SIMONSEN A. Quality control of the mitochondrion[J]. Dev Cell, 2021, 56(7): 881-905. DOI: 10.1016/j.devcel.2021.02.009.

BERGMAN R N, ADER M. Free fatty acids and pathogenesis of type 2 diabetes mellitus[J]. Trends Endocrinol Metab, 2000, 11(9): 351-356. DOI: 10.1016/S1043-2760(00)00323-4.

TORRES M J, RYAN T E, LIN C T, et al . Impact of 17β-estradiol on complex Ⅰ kinetics and H 2 O 2 production in liver and skeletal muscle mitochondria[J ] . J Biol Chem, 2018, 293(43): 16889-16898. DOI: 10.1074/jbc.RA118.005148..

SEKO D, FUJITA R, KITAJIMA Y, et al . Estrogen receptor β controls muscle growth and regeneration in young female mice[J ] . Stem Cell Rep, 2020, 15(3): 577-586. DOI: 10.1016/j.stemcr.2020.07.017.

JACKSON R L, GREIWE J S, SCHWEN R J. Emerging evidence of the health benefits of S-equol, an estrogen receptor β agonist[J]. Nutr Rev, 2011, 69(8): 432-448. DOI: 10.1111/j.1753-4887.2011.00400.x.

SETCHELL K D R, CLERICI C, LEPHART E D, et al . S-equol, a potent ligand for estrogen receptor beta, is the exclusive enantiomeric form of the soy isoflavone metabolite produced by human intestinal bacterialflora[J ] . Am J Clin Nutr, 2005, 81(5): 1072-1079. DOI: 10.1093/ajcn/81.5.1072.

YOSHINO M, NAKA A, SAKAMOTO Y, et al . Dietary isoflavone daidzein promotes Tfam expression that increases mitochondrial biogenesis in C2C12 muscle cells[J ] . J Nutr Biochem, 2015, 26(11): 1193-1199. DOI: 10.1016/j.jnutbio.2015.05.010.

杨宁宁, 王佑民, 陈冬, 等. 棕榈酸致大鼠L6肌细胞胰岛素抵抗与JNK1活化的关系[J]. 安徽医科大学学报, 2012, 47(3): 241-244. DOI: 10.3969/j.issn.1000-1492.2012.03.003.

YANG N N, WANG Y M, CHEN D, et al . Association of JNK1 activation with palmitic acid-induced insulin resistance in L6 rat muscle cells[J ] . Acta Univ Med Anhui, 2012, 47(3): 241-244. DOI: 10.3969/j.issn.1000-1492.2012.03.003.

IZZO A, MASSIMINO E, RICCARDI G, et al . A nar rative review on sarcopenia in type 2 diabetes mellitus: prevalence and associated factors[J ] . Nutrients, 2021, 13(1): 183. DOI: 10.3390/nu13010183.

DE FREITAS M M, DE OLIVEIRA V L P, GRASSI T, et al . Difference in sarcopenia prevalence and associated factors according to 2010 and 2018 European consensus (EWGSOP) in elderly patients with type 2 diabetes mellitus[J ] . Exp Gerontol, 2020, 132: 110835. DOI: 10.1016/j.exger.2020.110835.

LEE H, LIM J Y, CHOI S J. Oleate prevents palmitate-induced atrophy via modulation of mitochondrial ROS production in skeletal myotubes[J ] . Oxidative Med Cell Longev, 2017, 2017: 1-11. DOI: 10.1155/2017/2739721.

FEALY C E, GREVENDONK L, HOEKS J, et al . Skeletal muscle mitochondrial network dynamics in metabolic disorders and aging[J ] . Trends Mol Med, 2021, 27(11): 1033-1044. DOI: 10.1016/j.molmed.2021.07.013.

YU L, CHEN Y, TOOZE S A. Autophagy pathway: cellular and molecular mechanisms[J]. Autophagy, 2018, 14(2): 207-215. DOI: 10.1080/15548627.2017.1378838.

TANIDA I. Autophagosome formation and molecular mechanism of autophagy[J]. Antioxid Redox Signal, 2011, 14(11): 2201-2214. DOI: 10.1089/ars.2010.3482.

TEREŠAK P, LAPAO A N, SUBIC N, et al . Regulation of PRKN-independent mitophagy[J ] . Autophagy, 2022, 18(1): 24-39. DOI: 10.1080/15548627.2021.1888244.

PICKLES S, VIGIÉ P, YOULE R J. Mitophagy and quality control mechanisms in mitochondrial maintenance[J]. Curr Biol, 2018, 28(4): R170-R185. DOI: 10.1016/j.cub.2018.01.004.

KLINGE C M. Estrogenic control of mitochondrial function[J]. Redox Biol, 2020, 31: 101435. DOI: 10.1016/j.redox.2020.101435.

IKEDA K, HORIE-INOUE K, INOUE S. Functions of estrogen and estrogen receptor signaling on skeletal muscle[J]. J Steroid Biochem Mol Biol, 2019, 191: 105375. DOI: 10.1016/j.jsbmb.2019.105375.

KLINGE C M. Estrogenic control of mitochondrial function and biogenesis[J]. J Cell Biochem, 2008, 105(6): 1342-1351. DOI: 10.1002/jcb.21936.

RIBAS V, DREW B G, ZHOU Z Q, et al . Skeletal muscle action of e strogen receptor α is critical for the maintenance of mitochondrial function and metabolic homeostasis in females[J ] . Sci Transl Med, 2016, 8(334): 334ra54. DOI: 10.1126/scitranslmed.aad3815.

MAYO B, VÁZQUEZ L, FLÓREZ A B. Equol: a bacterial metabolite from the daidzein isoflavone and its presumed beneficial health effects[J]. Nutrients, 2019, 11(9): 2231. DOI: 10.3390/nu11092231.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

RANKL通过ROS介导HIF-1α下调促进自噬和破骨细胞分化

转录因子EB增强巨噬细胞自噬功能及逆转内毒素耐受的作用

Autophagy inducer spermidine improves brain injury after intracerebral hemorrhage

AKT1-mediated autophagy of hepatocellular carcinoma cells enhances cell sensitivity to ¹²⁵I seed irradiation

Effect of equol intervention on high-fat diet-induced non-alcoholic fatty liver disease in ovariectomized rats

Related Author

LI Songtao

SUN Jing

CHU Tongwei

YANG Ting

LIU Xin

JIANG Qingsong

WANG Yujie

SHI Xinhui

Related Institution

Department of Cerebrovascular Diseases， Second Affiliated Hospital of Zhengzhou University, Zhengzhou University

Academy of Medical Sciences, Zhengzhou University

Department of Nuclear Medicine， First Affiliated Hospital， Army Medical University （Third Military Medical University）

Department of Clinical Microbiology and Immunology, Faculty of Pharmacy and Medical Laboratory, Army Medical University (Third Military Medical University)

⁰