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

Hu,Yanwu; Liu,Kai; Yan,Mengtong; Zhang,Yang; Wang,Yadi; Ren,Liqun

doi:10.3892/mmr.2016.4813

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

Authors:
- Yanwu Hu
- Kai Liu
- Mengtong Yan
- Yang Zhang
- Yadi Wang
- Liqun Ren
View Affiliations

Affiliations: Department of Experimental Pharmacology and Toxicology, School of Pharmaceutical Science, Jilin University, Changchun, Jilin 130021, P.R. China
Published online on: January 27, 2016 https://doi.org/10.3892/mmr.2016.4813
Pages: 2899-2903

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Icariin, a flavonoid isolated from the traditional Chinese herbal medicine Epimedium brevicornum Maxim, has been shown to possess anti-inflammatory, anti‑oxidant and anti-atherosclerotic activities in vivo and in vitro. The aim of the present study was to investigate the effects of icariin on oxidized low‑density lipoprotein (ox-LDL)-induced proliferation of vascular smooth muscle cells (VSMCs) and the possible underlying mechanism. VSMCs were cultured and pre‑treated with various concentrations of icariin (0, 10, 20 or 40 µm) prior to stimulation by ox‑LDL (50 µg/ml). Cell proliferation was evaluated by an MTT assay. Flow cytometry was used to study the influence of icariin on the cell cycle. Proliferating cell nuclear antigen (PCNA) expression and phosphorylation levels of extracellular signal-regulated kinase (ERK)1/2 were detected by western blot analysis. The results indicated that icariin significantly inhibited ox‑LDL‑induced proliferation of VSMCs and phosphorylation of ERK1/2. Furthermore, icariin also blocked the ox‑LDL‑induced cell‑cycle progression at G1/S‑interphase and downregulated the expression of PCNA in VSMCs. In conclusion, the present study indicated for the first time that icariin reduced the amount of ox‑LDL‑induced proliferation of VSMCs through suppression of PCNA expression and inactivation of ERK1/2.

View Figures

View References

1	Dell'omo G, Penno G, Pucci L, Lucchesi D, Fotino C, Del Prato S and Pedrinelli R: ACE gene insertion/deletion polymorphism modulates capillary permeability in hypertension. Clin Sci (Lond). 111:3573–64. 2006. View Article : Google Scholar
2	Zhu F, Li C, Jin XP, Weng SX, Fan LL, Zheng Z, Li WL, Wang F, Wang WF, Hu XF, et al: Celastrol may have an anti-atherosclerosis effect in a rabbit experimental carotid atherosclerosis model. Int J Clin Exp Med. 7:1684–1691. 2014.PubMed/NCBI
3	Boehm M and Nabel EG: Cell cycle and cell migration: New pieces to the puzzle. Circulation. 103:2879–2881. 2001. View Article : Google Scholar : PubMed/NCBI
4	Chang WC, Yu YM, Chiang SY and Tseng CY: Ellagic acid suppresses oxidised low-density lipoprotein-induced aortic smooth muscle cell proliferation: Studies on the activation of extracellular signal-regulated kinase 1/2 and proliferating cell nuclear antigen expression. Br J Nutr. 99:709–714. 2008. View Article : Google Scholar : PubMed/NCBI
5	Xie F, Wu CF, Lai WP, Yang XJ, Cheung PY, Yao XS, Leung PC and Wong MS: The osteoprotective effect of Herba epimedii (HEP) extract in vivo and in vitro. Evid Based Complement Alternat Med. 2:353–361. 2005. View Article : Google Scholar : PubMed/NCBI
6	Hidaka S, Okamoto Y, Yamada Y, Kon Y and Kimura T: A Japanese herbal medicine, Chujo-to, has a beneficial effect on osteoporosis in rats. Phytother Res. 13:14–19. 1999. View Article : Google Scholar : PubMed/NCBI
7	Sakamoto S, Sassa S, Kudo H, Suzuki S, Mitamura T and Shinoda H: Preventive effects of a herbal medicine on bone loss in rats treated with a GnRH agonist. Eur J Endocrinol. 143:139–142. 2000. View Article : Google Scholar : PubMed/NCBI
8	Xiao-Hong D, Chang-Qin X, Jian-Hua H, Wen-Jiang Z and Bing S: Icariin delays homocysteine-induced endothelial cellular senescence involving activation of the PI3K/AKT-eNOS signaling pathway. Pharm Biol. 51:433–440. 2013. View Article : Google Scholar : PubMed/NCBI
9	Zeng KW, Fu H, Liu GX and Wang XM: Icariin attenuates lipo-polysaccharide-induced microglial activation and resultant death of neurons by inhibiting TAK1/IKK/NF-kappaB and JNK/p38 MAPK pathways. Int Immunopharmacol. 10:668–678. 2010. View Article : Google Scholar : PubMed/NCBI
10	Zhang G, Qin L, Sheng H, Yeung KW, Yeung HY, Cheung WH, Griffith J, Chan CW, Lee KM and Leung KS: Epimedium-derived phytoestrogen exert beneficial effect on preventing steroid-associated osteonecrosis in rabbits with inhibition of both thrombosis and lipid-deposition. Bone. 40:685–692. 2007. View Article : Google Scholar
11	Zhang DC, Liu JL, Ding YB, Xia JG and Chen GY: Icariin potentiates the antitumor activity of gemcitabine in gallbladder cancer by suppressing NF-κB. Acta Pharmacol Sin. 34:301–308. 2013. View Article : Google Scholar : PubMed/NCBI
12	Yap SP, Shen P, Li J, Lee LS and Yong EL: Molecular and pharmacodynamic properties of estrogenic extracts from the traditional Chinese medicinal herb, Epimedium. J Ethnopharmacol. 113:218–224. 2007. View Article : Google Scholar : PubMed/NCBI
13	Chen Y, Huang JH, Ning Y and Shen ZY: Icariin and its pharmaceutical efficacy: Research progress of molecular mechanism. J Chin Integr Med. 9:1179–1184. 2011.In Chinese. View Article : Google Scholar
14	Xu CQ, Liu BJ, Wu JF, Xu YC, Duan XH, Cao YX and Dong JC: Icariin attenuates LPS-induced acute inflammatory responses: Involvement of PI3K/Akt and NF-kappaB signaling pathway. Eur J Pharmacol. 642:146–153. 2010. View Article : Google Scholar : PubMed/NCBI
15	Xu HB and Huang ZQ: Vasorelaxant effects of icariin on isolated canine coronary artery. J Cardiovasc Pharmacol. 49:207–213. 2007. View Article : Google Scholar : PubMed/NCBI
16	Nurgeldiyeva MJ, Hojakuliyev BG and Muhammedov MB: Correlation of atherogenesis with an infection of Candida albicans. Int J Clin Exp Med. 7:2137–2143. 2014.PubMed/NCBI
17	Butcher MJ, Herre M, Ley K and Galkina E: Flow cytometry analysis of immune cells within murine aortas. J Vis Exp. pii: 2848. 2011. View Article : Google Scholar : PubMed/NCBI
18	Li Q, Wang Y, Li H, Shen G and Hu S: Ox-LDL influences peripheral Th17/Treg balance by modulating Treg apoptosis and Th17 proliferation in atherosclerotic cerebral infarction. Cell Physiol Biochem. 33:1849–1862. 2014. View Article : Google Scholar : PubMed/NCBI
19	Yang CM, Chien CS, Hsiao LD, Pan SL, Wang CC, Chiu CT and Lin CC: Mitogenic effect of oxidized low-density lipoprotein on vascular smooth muscle cells mediated by activation of Ras/Raf/MEK/MAPK pathway. Br J Pharmacol. 132:1531–1541. 2001. View Article : Google Scholar : PubMed/NCBI
20	Abate-Daga D, Hanada K, Davis JL, Yang JC, Rosenberg SA and Morgan RA: Expression profiling of TCR-engineered T cells demonstrates overexpression of multiple inhibitory receptors in persisting lymphocytes. Blood. 122:1399–1410. 2013. View Article : Google Scholar : PubMed/NCBI
21	Tian R, Li Y and Gao M: Shikonin causes cell cycle arrest and induces apoptosis by regulating the EGFR/NF-κB signaling pathway in human epidermoid carcinoma A431 cells. Biosci Rep. 35:pii: e00189. 2015. View Article : Google Scholar
22	Li QY, Zhu YF, Zhang M, Chen L, Zhang Z, Du YL, Ren GQ, Tang JM, Zhong MK and Shi XJ: Chlorogenic acid inhibits hypoxia-induced pulmonary artery smooth muscle cells proliferation via c-Src and Shc/Grb2/ERK2 signaling pathway. Eur J Pharmacol. 751:81–88. 2015. View Article : Google Scholar : PubMed/NCBI
23	Lu W, Du S and Wang J: Berberine inhibits the proliferation of prostate cancer cells and induces G₀/G₁ or G₂/M phase arrest at different concentrations. Mol Med Rep. 11:3920–3924. 2015.PubMed/NCBI
24	Elledge SJ: Cell cycle checkpoints: Preventing an identity crisis. Science. 274:1664–1672. 1996. View Article : Google Scholar : PubMed/NCBI
25	Kim JY, Kim KH, Lee WR, An HJ, Lee SJ, Han SM, Lee KG, Park YY, Kim KS and Lee YS: Apamin inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and migration through suppressions of activated Akt and Erk signaling pathway. Vascul Pharmacol. 70:8–14. 2015. View Article : Google Scholar : PubMed/NCBI
26	He H, Tan CK, Downey KM and So AG: A tumor necrosis factor alpha- and interleukin 6-inducible protein that interacts with the small subunit of DNA polymerase delta and proliferating cell nuclear antigen. Proc Natl Acad Sci USA. 98:11979–11984. 2001. View Article : Google Scholar : PubMed/NCBI
27	Li YY, Wang L and Lu CD: An E2F site in the 5′-promoter region contributes to serum-dependent up-regulation of the human proliferating cell nuclear antigen gene. Febs Lett. 544:112–118. 2003. View Article : Google Scholar : PubMed/NCBI
28	Roskoski R Jr: ERK1/2 MAP kinases: Structure, function and regulation. Pharmacol Res. 66:105–143. 2012. View Article : Google Scholar : PubMed/NCBI
29	Zhang Z, Zhang M, Li Y, Liu S, Ping S, Wang J, Ning F, Xie F and Li C: Simvastatin inhibits the additive activation of ERK1/2 and proliferation of rat vascular smooth muscle cells induced by combined mechanical stress and oxLDL through LOX-1 pathway. Cell Signal. 25:332–340. 2013. View Article : Google Scholar
30	Farrokhi E, Samani KG and Chaleshtori MH: Oxidized low-density lipoprotein increases bone sialoprotein expression in vascular smooth muscle cells via runt-related transcription factor 2. Cell Signal. 349:240–243. 2015.