CC BY 4.0 · Chinese medicine and natural products 2023; 03(01): e41-e50
DOI: 10.1055/s-0043-1764132
Original Article

Wenyang-Yiqi Granule Suppresses Oxygen-Glucose Deprivation-Induced Cardiomyocyte Autophagy Through Mammalian Target of Rapamycin Activation in H9c2 Cells

Shuibo Gao
1   Laboratory of Cell Imaging, Second School of Clinical Medicine, Henan University of Chinese Medicine, Zhengzhou, Henan, China
Xiaofang Yu
2   Institute of Cardiovascular Disease, Henan University of Chinese Medicine, Zhengzhou, Henan, China
Lihua Han
2   Institute of Cardiovascular Disease, Henan University of Chinese Medicine, Zhengzhou, Henan, China
Hong Wu
1   Laboratory of Cell Imaging, Second School of Clinical Medicine, Henan University of Chinese Medicine, Zhengzhou, Henan, China
2   Institute of Cardiovascular Disease, Henan University of Chinese Medicine, Zhengzhou, Henan, China
› Author Affiliations
Funding This work was funded by the National Natural Science Foundation of China (81373610, 81673800).


Background Wenyang-Yiqi Granule (WYYQ) is a four-component herbal formula, widely used to treat heart failure in China. It is known to regulate autophagy, but the mechanism(s) are unknown.

Methods H9c2 cells were treated with WYYQ for 24 hours prior to oxygen-glucose deprivation (OGD). Expressions of the autophagy markers Beclin-1 and light chain 3 (LC3) were evaluated via quantitative polymerase chain reaction analysis. Protein levels of Beclin-1, LC3, p62, and mammalian targets of rapamycin (mTOR) were determined by Western blot analysis. Transmission electron microscopy was used to explore the effects of WYYQ on autophagosome formation.

Results Treatment with WYYQ dramatically restrained OGD-induced autophagy, which was characterized by an inhibition of Beclin-1 and increased LC3 mRNA expression. In addition, WYYQ decreased the expression of Beclin-1 and the ratio of LC3-II/LC3-I; however, the abundance of p62 was enhanced at the protein level. Manipulation of the LC3-II/LC3-I ratio, p62 abundance, and autophagosome formation in response to WYYQ were associated with mTOR activity.

Conclusions These findings show that WYYQ plays a protective role during hypoxic-ischemic stress through the suppression of excessive autophagy, which may be partially explained by its effects on mTOR. These data provide novel insight into the cardioprotective effects of WYYQ during cardiomyocyte autophagy.

CRediT Authorship Contribution Statement

L.H. and H.W. were responsible for conceptualization, methodology, and formal analysis. S.G., X.Y., and H.W. were responsible for funding acquisition, writing original draft, and writing—review and editing.

Publication History

Received: 07 September 2022

Accepted: 20 October 2022

Article published online:
14 March 2023

© 2023. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

  • References

  • 1 Papanagnou ED, Gumeni S, Sklirou AD. et al. Autophagy activation can partially rescue proteasome dysfunction-mediated cardiac toxicity. Aging Cell 2022; 21 (11) e13715
  • 2 Cui X, Wang Y, Liu H, Shi M, Wang J, Wang Y. The molecular mechanisms of defective copper metabolism in diabetic cardiomyopathy. Oxid Med Cell Longev 2022; 2022: 5418376
  • 3 Li Z, Wang J, Yang X. Functions of autophagy in pathological cardiac hypertrophy. Int J Biol Sci 2015; 11 (06) 672-678
  • 4 Gatica D, Chiong M, Lavandero S, Klionsky DJ. Molecular mechanisms of autophagy in the cardiovascular system. Circ Res 2015; 116 (03) 456-467
  • 5 Peng L, Zhuang X, Liao L. et al. Changes in cell autophagy and apoptosis during age-related left ventricular remodeling in mice and their potential mechanisms. Biochem Biophys Res Commun 2013; 430 (02) 822-826
  • 6 Yang M, Ji X, Zuo Z. Relationships between the toxicities of Radix aconiti lateralis preparata (Fuzi) and the toxicokinetics of its main diester-diterpenoid alkaloids. Toxins (Basel) 2018; 10 (10) 391
  • 7 Sun L, Liu LN, Li JC. et al. The essential oil from the twigs of Cinnamomum cassia Presl inhibits oxytocin-induced uterine contraction in vitro and in vivo. J Ethnopharmacol 2017; 206: 107-114
  • 8 Shahzad M, Shabbir A, Wojcikowski K, Wohlmuth H, Gobe GC. The antioxidant effects of Radix astragali (Astragalus membranaceus and related species) in protecting tissues from injury and disease. Curr Drug Targets 2016; 17 (12) 1331-1340
  • 9 Li J, Liu S, Wang J. et al. Fungal elicitors enhance ginsenosides biosynthesis, expression of functional genes as well as signal molecules accumulation in adventitious roots of Panax ginseng C. A. Mey. J Biotechnol 2016; 239: 106-114
  • 10 Han L, Wang Z, Chai S, Xing G, Wang H. Effects of comparison of warming yang prescription with invigorating qi prescription on renin, angiotensin II, and idosterone at different time phase in rats with heart failure after myocardial infarction. CJTCMP 2015; 30 (11) 4103-4105
  • 11 Dai DF, Rabinovitch P. Mitochondrial oxidative stress mediates induction of autophagy and hypertrophy in angiotensin-II treated mouse hearts. Autophagy 2011; 7 (08) 917-918
  • 12 Ichihara A. (Pro)renin receptor and autophagy in podocytes. Autophagy 2012; 8 (02) 271-272
  • 13 Porrello ER, Delbridge LM. Cardiomyocyte autophagy is regulated by angiotensin II type 1 and type 2 receptors. Autophagy 2009; 5 (08) 1215-1216
  • 14 Zheng S, Han F, Shi Y, Wen L, Han D. Single-prolonged-stress-induced changes in autophagy-related proteins beclin-1, LC3, and p62 in the medial prefrontal cortex of rats with post-traumatic stress disorder. J Mol Neurosci 2017; 62 (01) 43-54
  • 15 Masuda GO, Yashiro M, Kitayama K. et al. Clinicopathological correlations of autophagy-related proteins LC3, beclin 1 and p62 in gastric cancer. Anticancer Res 2016; 36 (01) 129-136
  • 16 Ma LL, Ma X, Kong FJ. et al. Mammalian target of rapamycin inhibition attenuates myocardial ischaemia-reperfusion injury in hypertrophic heart. J Cell Mol Med 2018; 22 (03) 1708-1719
  • 17 Hang P, Zhao J, Su Z. et al. Choline inhibits ischemia-reperfusion-induced cardiomyocyte autophagy in rat myocardium by activating Akt/mTOR signaling. Cell Physiol Biochem 2018; 45 (05) 2136-2144
  • 18 Weinberg MA. RES-529: a PI3K/AKT/mTOR pathway inhibitor that dissociates the mTORC1 and mTORC2 complexes. Anticancer Drugs 2016; 27 (06) 475-487
  • 19 Noda T, Ohsumi Y. Tor, a phosphatidylinositol kinase homologue, controls autophagy in yeast. J Biol Chem 1998; 273 (07) 3963-3966
  • 20 Watanabe-Asano T, Kuma A, Mizushima N. Cycloheximide inhibits starvation-induced autophagy through mTORC1 activation. Biochem Biophys Res Commun 2014; 445 (02) 334-339
  • 21 Settembre C, Fraldi A, Medina DL, Ballabio A. Signals from the lysosome: a control centre for cellular clearance and energy metabolism. Nat Rev Mol Cell Biol 2013; 14 (05) 283-296
  • 22 Chiang MH, Liang CJ, Liu CW. et al. Aliskiren improves ischemia- and oxygen glucose deprivation-induced cardiac injury through activation of autophagy and AMP-activated protein kinase. Front Pharmacol 2017; 8: 819
  • 23 Guan ZF, Zhang XM, Tao YH. et al. EGb761 improves the cognitive function of elderly db/db-/- diabetic mice by regulating the beclin-1 and NF-κB signaling pathways. Metab Brain Dis 2018; 33 (06) 1887-1897
  • 24 Wu H, Gao S, Fu M, Sakurai T, Terakawa S. Fucoidan inhibits Ca2+ responses induced by a wide spectrum of agonists for G‑protein‑coupled receptors. Mol Med Rep 2018; 17 (01) 1428-1436
  • 25 Wu H, Gao H, Gao S. et al. A Chinese 4-herb formula, Yiqi-Huoxue granule, alleviates H2O2-induced apoptosis by upregulating uncoupling protein 2 in H9c2 cells. Phytomedicine 2019; 53: 171-181
  • 26 Moliner-Abós C, Rivas-Lasarte M, Pamies Besora J. et al. Sacubitril/valsartan in real-life practice: experience in patients with advanced heart failure and systematic review. Cardiovasc Drugs Ther 2019; 33 (03) 307-314
  • 27 Ghosh R, Gillaspie JJ, Campbell KS, Symons JD, Boudina S, Pattison JS. Chaperone-mediated autophagy protects cardiomyocytes against hypoxic-cell death. Am J Physiol Cell Physiol 2022; 323 (05) C1555-C1575
  • 28 Wang S, Yao T, Deng F. et al. LncRNA MALAT1 promotes oxygen-glucose deprivation and reoxygenation induced cardiomyocytes injury through sponging miR-20b to enhance beclin1-mediated autophagy. Cardiovasc Drugs Ther 2019; 33 (06) 675-686
  • 29 Calis S, Dogan B, Durdagi S. et al. A novel BH3 mimetic Bcl-2 inhibitor promotes autophagic cell death and reduces in vivo Glioblastoma tumor growth. Cell Death Discov 2022; 8 (01) 433
  • 30 Saxton RA, Sabatini DM. mTOR signaling in growth, metabolism, and disease. Cell 2017; 168 (06) 960-976
  • 31 Schüttler D, Clauss S, Weckbach LT, Brunner S. Molecular mechanisms of cardiac remodeling and regeneration in physical exercise. Cells 2019; 8 (10) 1128
  • 32 Shende P, Plaisance I, Morandi C. et al. Cardiac raptor ablation impairs adaptive hypertrophy, alters metabolic gene expression, and causes heart failure in mice. Circulation 2011; 123 (10) 1073-1082
  • 33 Ramos FJ, Chen SC, Garelick MG. et al. Rapamycin reverses elevated mTORC1 signaling in lamin A/C-deficient mice, rescues cardiac and skeletal muscle function, and extends survival. Sci Transl Med 2012; 4 (144): 144ra103
  • 34 Wang H, Hua J, Chen S, Chen Y. SERPINB1 overexpression protects myocardial damage induced by acute myocardial infarction through AMPK/mTOR pathway. BMC Cardiovasc Disord 2022; 22 (01) 107
  • 35 Zhu H, Tannous P, Johnstone JL. et al. Cardiac autophagy is a maladaptive response to hemodynamic stress. J Clin Invest 2007; 117 (07) 1782-1793
  • 36 Zou Y, Lin L, Ye Y. et al. Qiliqiangxin inhibits the development of cardiac hypertrophy, remodeling, and dysfunction during 4 weeks of pressure overload in mice. J Cardiovasc Pharmacol 2012; 59 (03) 268-280
  • 37 Shi X, Zhu H, Zhang Y, Zhou M, Tang D, Zhang H. XuefuZhuyu decoction protected cardiomyocytes against hypoxia/reoxygenation injury by inhibiting autophagy. BMC Complement Altern Med 2017; 17 (01) 325
  • 38 Yu X, Zhao XD, Bao RQ, Yu JY, Zhang GX, Chen JW. The modified Yi qi decoction protects cardiac ischemia-reperfusion induced injury in rats. BMC Complement Altern Med 2017; 17 (01) 330
  • 39 Cui H, Li X, Li N. et al. Induction of autophagy by Tongxinluo through the MEK/ERK pathway protects human cardiac microvascular endothelial cells from hypoxia/reoxygenation injury. J Cardiovasc Pharmacol 2014; 64 (02) 180-190
  • 40 Liu L, Wu Y, Huang X. Orientin protects myocardial cells against hypoxia-reoxygenation injury through induction of autophagy. Eur J Pharmacol 2016; 776: 90-98
  • 41 Zhao H, Zhang M, Zhou F. et al. Cinnamaldehyde ameliorates LPS-induced cardiac dysfunction via TLR4-NOX4 pathway: the regulation of autophagy and ROS production. J Mol Cell Cardiol 2016; 101: 11-24
  • 42 Yin B, Hou XW, Lu ML. Astragaloside IV attenuates myocardial ischemia/reperfusion injury in rats via inhibition of calcium-sensing receptor-mediated apoptotic signaling pathways. Acta Pharmacol Sin 2019; 40 (05) 599-607
  • 43 Chen W, Sun Q, Ju J. et al. Effect of Astragalus polysaccharides on cardiac dysfunction in db/db mice with respect to oxidant stress. BioMed Res Int 2018; 2018: 8359013
  • 44 Sun S, Yang S, An N. et al. Astragalus polysaccharides inhibits cardiomyocyte apoptosis during diabetic cardiomyopathy via the endoplasmic reticulum stress pathway. J Ethnopharmacol 2019; 238: 111857
  • 45 Chen X, Wang Q, Shao M. et al. Ginsenoside Rb3 regulates energy metabolism and apoptosis in cardiomyocytes via activating PPARα pathway. Biomed Pharmacother 2019; 120: 109487