Bioactive molecule Icariin inhibited proliferation while enhanced apoptosis and autophagy of rat airway smooth muscle cells in vitro

  • Lihong Yang
  • Bo Xiao
  • Lixia Hou
  • Guiming Zhou
  • Biwen Mo
  • Dong YaoEmail author
Original Article


Icariin is the main active compound extracted from epimedium Flavonoids (EFs) and involved in regulation of cell behaviors (proliferation, apoptosis, and autophagy etc.) for many cell types, but the effect of Icariin on airway smooth muscle cells (ASMCs) is still unknown. The aim of the present study is to examine the role of Icariin on rat ASMCs proliferation, apoptosis and autophagy. CKK8 assay showed that Icariin inhibited rat ASMCs growth in dose-time-dependent manner, and the flow cytometry assay showed that the Icariin interfered with ASMCs cell cycle, when treated with Icariin, S phase shortened while G2 phase extended, cyclin E1 and cyclinA1 gene and protein expression decreased. Next apoptosis was detected, Flow cytometry and TdTmediated dUTP Nick-End Labeling (TUNEL) assay showed that Icariin promoted ASMCs apoptosis, and enhanced apoptosis protein cleaved-caspase-3 expression. Finally, it was found Icariin induced rat ASMCs autophagy, with enhancement expression of autophagy marker LC3 II. In conclusion, Icariin inhibited ASMCs proliferation while promoted apoptosis and autophagy, revealing its potential role in treatment of airway remodeling in asthma.


Icariin Airway smooth muscle cells Proliferation Apoptosis Autophagy 



We thank the staffs of Science Laboratory Center of Guilin Medical University for technical supports.


This study was supported by National Natural Science Foundation of China (Grant Nos. 81560005, 81760007) and Natural Science Foundation of Guangxi Province (2016GXNSFBA380003)

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. Abrams JM, White MA (2004) Coordination of cell death and the cell cycle: linking proliferation to death through private and communal couplers. Curr Opin Cell Biol 16:634–638CrossRefGoogle Scholar
  2. Chen MF, Qi L, Li Y, Zu XB, Dai YQ, Zhang P (2010) Icaritin induces growth inhibition and apoptosis of human prostatic smooth muscle cells in an estrogen receptor-independent manner. Amino Acids 38:1505–1513CrossRefGoogle Scholar
  3. Du WJ, Dong JC, Cai C (2011) Effects of icariin on Bcl-2 and Bax protein expressions and eosinophils apoptosis in bronchial asthmatic mice. Zhongguo Zhong Xi Yi Jie He Za Zhi 31:1248–1253PubMedGoogle Scholar
  4. Gu ZF, Zhang ZT, Wang JY, Xu BB (2017) Icariin exerts inhibitory effects on the growth and metastasis of KYSE70 human esophageal carcinoma cells via PI3 K/AKT and STAT3 pathways. Environ Toxicol Pharmacol 54:7CrossRefGoogle Scholar
  5. He F, Li B, Zhao Z, Zhou Y, Hu G, Zou W, Hong W, Zou Y, Jiang C, Zhao D (2014) The pro-proliferative effects of nicotine and its underlying mechanism on rat airway smooth muscle cells. PLoS ONE 9:e93508CrossRefGoogle Scholar
  6. Henglein B, Chenivesse X, Wang J, Eick D, Bréchot C (1994) Structure and cell cycle-regulated transcription of the human cyclin A gene. Proc Natl Acad Sci USA 91:5490–5494CrossRefGoogle Scholar
  7. Hinds PW, Mittnacht S, Dulic V, Arnold A, Reed SI, Weinberg RA (1992) Regulation of retinoblastoma protein functions by ectopic expression of human cyclins. Cell 70:993–1006CrossRefGoogle Scholar
  8. Hirst SJ (1996) Airway smooth muscle cell culture: application to studies of airway wall remodelling and phenotype plasticity in asthma. Eur Respir J 9:808–820CrossRefGoogle Scholar
  9. Hu Y, Liu K, Yan M, Zhang Y, Wang Y, Ren L (2016) Icariin inhibits oxidized low-density lipoprotein-induced proliferation of vascular smooth muscle cells by suppressing activation of extracellular signal-regulated kinase 1/2 and expression of proliferating cell nuclear antigen. Mol Med Rep 13:2899–2903CrossRefGoogle Scholar
  10. Hubbi ME, Semenza GL (2015) An essential role for chaperone-mediated autophagy in cell cycle progression. Autophagy 11:850–851CrossRefGoogle Scholar
  11. Lambert M, Thybo C, Lykkeboe S, Rasmussen L, Frette X, Christensen L, Jeppesen P (2017) Combined bioavailable isoflavones and probiotics improve bone status and estrogen metabolism in postmenopausal osteopenic women: a randomized controlled trial. Am J Clin Nutr 106:909–920PubMedGoogle Scholar
  12. Li J, Gang D, Yu X, Hu Y, Yue Y, Cheng W, Pan X, Zhang P (2013) Genistein: the potential for efficacy in rheumatoid arthritis. Clin Rheumatol 32:535–540CrossRefGoogle Scholar
  13. Li ZJ, Yao C, Liu SF, Chen L, Xi YM, Zhang W, Zhang GS (2014) Cytotoxic effect of icaritin and its mechanisms in inducing apoptosis in human burkitt lymphoma cell line. Biomed Res Int 2014:391512PubMedPubMedCentralGoogle Scholar
  14. Li Z, Zhang Y, Chen L, Li H (2018) The dietary compound luteolin inhibits pancreatic cancer growth by targeting BCL-2. Food Funct 9:3018–3027CrossRefGoogle Scholar
  15. Mariño G, Nisosantano M, Baehrecke EH, Kroemer G (2014) Self-consumption: the interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol 15:81–94CrossRefGoogle Scholar
  16. Oak C, Khalifa A, Isali I, Bhaskaran N, Walker E, Shukla S (2018) Diosmetin suppresses human prostate cancer cell proliferation through the induction of apoptosis and cell cycle arrest. Int J Oncol 53:835–843PubMedPubMedCentralGoogle Scholar
  17. Pagano M, Pepperkok R, Verde F, Ansorge W, Draetta G (1992) Cyclin A is required at two points in the human cell cycle. EMBO J 11:961–971CrossRefGoogle Scholar
  18. Phillips B (1994) Cyclin E controls S phase progression and its down-regulation during Drosophila embryogenesis is required for the arrest of cell proliferation. Cell 77:107–120CrossRefGoogle Scholar
  19. Prakash YS (2013) Airway smooth muscle in airway reactivity and remodeling: what have we learned? Am J Physiol Lung Cell Mol Physiol 305:912–933CrossRefGoogle Scholar
  20. Qiao J, Sun S, Yuan L, Wang J (2017) Effects of icariin on asthma mouse model are associated with regulation of prostaglandin D2 level. Allergol Immunopathol (Madr) 45:567–572CrossRefGoogle Scholar
  21. Qin L, Zhang G, Sheng H, Wang XL, Wang YX, Yeung KW, Griffith JF, Li ZR, Leung KS, Yao XS (2008) Phytoestrogenic compounds for prevention of steroid-associated osteonecrosis. J Musculoskelet Neuronal Interact 8:18–21PubMedGoogle Scholar
  22. Shi DB, Li XX, Zheng HT, Li DW, Cai GX, Peng JJ, Gu WL, Guan ZQ, Xu Y, Cai SJ (2014) Icariin-mediated inhibition of NF-κB activity enhances the in vitro and in vivo antitumour effect of 5-fluorouracil in colorectal cancer. Cell Biochem Biophys 69:523–530CrossRefGoogle Scholar
  23. Song L, Zhao J, Zhang X, Li H, Zhou Y (2013) Icariin induces osteoblast proliferation, differentiation and mineralization through estrogen receptor-mediated ERK and JNK signal activation. Eur J Pharmacol 714:15–22CrossRefGoogle Scholar
  24. Song YH, Cai H, Zhao ZM, Chang WJ, Gu N, Cao SP, Wu ML (2016) Icariin attenuated oxidative stress induced-cardiac apoptosis by mitochondria protection and ERK activation. Biomed Pharmacother 83:1089–1094CrossRefGoogle Scholar
  25. Sun Y, Sun XH, Fan WJ, Jiang XM, Li AW (2016) Icariin induces S-phase arrest and apoptosis in medulloblastoma cells. Cell Mol Biol (Noisy-le-grand) 62:123–129Google Scholar
  26. Suzan R, Sander VDH (2016) Coordinating cell proliferation and differentiation: antagonism between cell cycle regulators and cell type-specific gene expression. Cell Cycle 15:196–212CrossRefGoogle Scholar
  27. Vermeulen K, Berneman ZN, Bockstaele DRV (2003) Cell cycle and apoptosis. Cell Prolif 36:165–175CrossRefGoogle Scholar
  28. Wang XF, Wang J (2014) Icaritin suppresses the proliferation of human osteosarcoma cells in vitro by increasing apoptosis and decreasing MMP expression. Acta Pharmacol Sin 35:531–539CrossRefGoogle Scholar
  29. Wang L, Zhang L, Chen ZB, Wu JY, Zhang X, Xu Y (2009) Icariin enhances neuronal survival after oxygen and glucose deprivation by increasing SIRT1. Eur J Pharmacol 609:40–44CrossRefGoogle Scholar
  30. Wang Q et al (2011) Icariin induces apoptosis in mouse MLTC-10 Leydig tumor cells through activation of the mitochondrial pathway and down-regulation of the expression of piwil4 International. J Oncol 39:973–980Google Scholar
  31. Wang ZM, Song N, Ren YL (2015) Anti-proliferative and cytoskeleton-disruptive effects of icariin on HepG2 cells. Mol Med Rep 12:6815–6820CrossRefGoogle Scholar
  32. Wiegman CH, Michaeloudes C, Haji G, Narang P, Clarke CJ, Russell KE, Bao W, Pavlidis S, Barnes PJ, Kanerva J (2015) Oxidative stress–induced mitochondrial dysfunction drives inflammation and airway smooth muscle remodeling in patients with chronic obstructive pulmonary disease. J Allergy Clin Immunol 136:769–780CrossRefGoogle Scholar
  33. Wright DB, Trian T, Siddiqui S, Pascoe CD, Johnson JR, Dekkers BG, Dakshinamurti S, Bagchi R, Burgess JK, Kanabar V (2013) Phenotype modulation of airway smooth muscle in asthma. Pulm Pharmacol Ther 26:42–49CrossRefGoogle Scholar
  34. Xu CQ, Le JJ, Duan XH, Du WJ, Liu BJ, Wu JF, Cao YX, Dong JC (2011) Molecular mechanism of icariin on rat asthmatic model. Chin Med J (Engl) 124:2899–2906Google Scholar
  35. Yang R, Muller C, Huynh V, Fung YK, Yee AS, Koeffler HP (1999) Functions of cyclin A1 in the cell cycle and its interactions with transcription factor E2F-1 and the Rb family of proteins. Mol Cell Biol 19:2400–2407CrossRefGoogle Scholar
  36. Zhou C, Chen Z, Lu X, Wu H, Yang Q, Xu D (2016) Icaritin activates JNK-dependent mPTP necrosis pathway in colorectal cancer cells. Tumour Biol 37:3135–3144CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Respiratory & Critical Care MedicineThe Affiliated Hospital of Guilin Medical UniversityGuilinChina
  2. 2.Key Laboratory of Respiratory Diseases of Colleges and Universities Affiliated Education Department of Guangxi Zhuang Autonomous RegionGuilinChina
  3. 3.Laboratory of Respiratory Disease, The Affiliated Hospital of Guilin Medical UniversityGuilinChina

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