Cucurbitacins mitigate vascular neointimal hyperplasia by suppressing cyclin A2 expression and inhibiting VSMC proliferation

Abstract

Background: Restenosis frequently occurs after percutaneous angioplasty in patients with vascular occlusion and seriously threatens their health. Substantial evidence has revealed that preventing vascular smooth muscle cell proliferation using a drug-eluting stent is an effective approach to improve restenosis. Cucurbitacins have been demonstrated to exert an anti-proliferation effect in various tumors and a hypotensive effect. This study aims to investigate the role of cucurbitacins extracted from Cucumis melo L. (CuECs) and cucurbitacin B (CuB) on restenosis.

Methods: C57BL/6 mice were subjected to left carotid artery ligation and subcutaneously injected with CuECs or CuB for 4 weeks. Hematoxylin–Eosin, immunofluorescence and immunohistochemistry staining were used to evaluate the effect of CuECs and CuB on neointimal hyperplasia. Western blot, real-time PCR, flow cytometry analysis, EdU staining and cellular immunofluorescence assay were employed to measure the effects of CuECs and CuB on cell proliferation and the cell cycle in vitro. The potential interactions of CuECs with cyclin A2 were performed by molecular docking.

Results: The results demonstrated that both CuECs and CuB exhibited significant inhibitory effects on neointimal hyperplasia and proliferation of vascular smooth muscle cells. Furthermore, CuECs and CuB mediated cell cycle arrest at the S phase. Autodocking analysis demonstrated that CuB, CuD, CuE and CuI had high binding energy for cyclin A2. Our study also showed that CuECs and CuB dramatically inhibited FBS-induced cyclin A2 expression. Moreover, the expression of cyclin A2 in CuEC- and CuB-treated neointima was downregulated.

Conclusions: CuECs, especially CuB, exert an anti-proliferation effect in VSMCs and may be potential drugs to prevent restenosis.

Keywords

CuB / Cucurbitacin / cyclin A2 / restenosis / vascular smooth muscle cell

References

[1] Malakar AK, Choudhury D, Halder B, Paul P, Uddin A, Chakraborty S. A review on coronary artery disease, its risk factors, and therapeutics. J Cell Physiol. 2019;234:16812-16823.

[2] Bauersachs R, Zeymer U, Brière JB, Marre C, Bowrin K, Huelsebeck M. Burden of coronary artery disease and peripheral artery disease: a literature review. Cardiovasc Ther. 2019;2019:8295054.

[3] Simard T, Hibbert B, Ramirez FD, Froeschl M, Chen YX, O’Brien ER. The evolution of coronary stents: a brief review. Can J Cardiol. 2014;30:35-45.

[4] Zhang X, Wang Q, Wang X, et al. Tanshinone IIA protects against heart failure post-myocardial infarction via AMPKs/mTOR-dependent autophagy pathway. Biomed Pharmacother. 2019;112:108599.

[5] Pei F, Pei H, Su C, et al. Fisetin alleviates Neointimal hyperplasia via PPARγ/PON2 Antioxidative pathway in SHR rat artery injury model. Oxidative Med Cell Longev. 2021;2021:6625517.

[6] Mills B, Robb T, Larson DF. Intimal hyperplasia: slow but deadly. Perfusion. 2012;27:520-528.

[7] Liu P, Xiang Y, Liu X, et al. Cucurbitacin B induces the lysosomal degradation of EGFR and suppresses the CIP2A/PP2A/Akt signaling Axis in Gefitinib-resistant non-small cell lung cancer. Molecules (Basel, Switzerland). 2019;24:647.

[8] Guo H, Kuang S, Song QL, Liu M, Sun XX, Yu Q. Cucurbitacin I inhibits STAT3, but enhances STAT1 signaling in human cancer cells in vitro through disrupting Actin filaments. Acta Pharmacol Sin. 2018;39:425-437.

[9] Zhang YZ, Wang CF, Zhang LF. Cucurbitacin D impedes gastric cancer cell survival via activation of the iNOS/NO and inhibition of the Akt signalling pathway. Oncol Rep. 2018;39:2595-2603.

[10] Cheng AC, Hsu YC, Tsai CC. The effects of cucurbitacin E on GADD45β-trigger G2/M arrest and JNK-independent pathway in brain cancer cells. J Cell Mol Med. 2019;23:3512-3519.

[11] Peng Y, Chen T, Luo L, et al. Isoforskolin and Cucurbitacin IIa promote the expression of anti-inflammatory regulatory factor SIGIRR in human macrophages stimulated with Borrelia burgdorferi basic membrane protein a. Int Immunopharmacol. 2020;88:106914.

[12] Silvestre GFG, de Lucena RP, da Silva AH. Cucurbitacins and the immune system: update in research on anti-inflammatory, antioxidant, and immunomodulatory mechanisms. Curr Med Chem. 2022;29:3774-3789.

[13] Alghasham AA. Cucurbitacins-a promising target for cancer therapy. Int J Health Sci (Qassim). 2013;7:77-89.

[14] Yuan RQ, Qian L, Yun WJ, et al. Cucurbitacins extracted from Cucumis melo L. (CuEC) exert a hypotensive effect via regulating vascular tone. Hypertens Res. 2019;42:1152-1161.

[15] Martínez-Alonso D, Malumbres M. Mammalian cell cycle cyclins. Semin Cell Dev Biol. 2020;107:28-35.

[16] Kozicka Z, Suchyta DJ, Focht V, et al. Design principles for cyclin K molecular glue degraders. Nat Chem Biol. 2024;20:93-102.

[17] Cui Y, Chen W, Du L, He Z. OIP5 interacts with NCK2 to mediate human Spermatogonial stem cell self-renewal and apoptosis through cell cyclins and cycle progression and its abnormality is correlated with male infertility. Research (Washington, DC). 2023;6:0162.

[18] Yasmeen A, Berdel WE, Serve H, Müller-Tidow C. E-and A-type cyclins as markers for cancer diagnosis and prognosis. Expert Rev Mol Diagn. 2003;3:617-633.

[19] Liu M, Gomez D. Smooth muscle cell phenotypic diversity. Arterioscler Thromb Vasc Biol. 2019;39:1715-1723.

[20] Sandison ME, Dempster J, McCarron JG. The transition of smooth muscle cells from a contractile to a migratory, phagocytic phenotype: direct demonstration of phenotypic modulation. J Physiol. 2016;594:6189-6209.

[21] Díez-Juan A, Andrés V. Coordinate control of proliferation and migration by the p27Kip1/cyclin-dependent kinase/retinoblastoma pathway in vascular smooth muscle cells and fibroblasts. Circ Res. 2003;92:402-410.

[22] Karpurapu M, Wang D, Singh NK, Li Q, Rao GN. NFATc1 targets cyclin a in the regulation of vascular smooth muscle cell multiplication during restenosis. J Biol Chem. 2008;283:26577-26590.

[23] Tammali R, Saxena A, Srivastava SK, Ramana KV. Aldose reductase regulates vascular smooth muscle cell proliferation by modulating G1/S phase transition of cell cycle. Endocrinology. 2010;151:2140-2150.

[24] Guan H, Gao L, Zhu L, et al. Apigenin attenuates neointima formation via suppression of vascular smooth muscle cell phenotypic transformation. J Cell Biochem. 2012;113:1198-1207.

[25] McPherson JA, Barringhaus KG, Bishop GG, et al. Adenosine a(2A) receptor stimulation reduces inflammation and neointimal growth in a murine carotid ligation model. Arterioscler Thromb Vasc Biol. 2001;21:791-796.

[26] Kamimura M, Bea F, Akizawa T, Katus HA, Kreuzer J, Viedt C. Platelet-derived growth factor induces tissue factor expression in vascular smooth muscle cells via activation of Egr-1. Hypertension (Dallas, Tex:1979). 2004;44:944-951.

[27] Chen Y, Chen Y, Jiang X, et al. Vascular adventitial fibroblasts-derived FGF10 promotes vascular smooth muscle cells proliferation and migration in vitro and the Neointima formation in vivo. J Inflamm Res. 2021;14:2207-2223.

[28] Heo KS, Cushman HJ, Akaike M, et al. ERK5 activation in macrophages promotes efferocytosis and inhibits atherosclerosis. Circulation. 2014;130:180-191.

[29] Park HS, Quan KT, Han JH, et al. Rubiarbonone C inhibits platelet-derived growth factor-induced proliferation and migration of vascular smooth muscle cells through the focal adhesion kinase, MAPK and STAT3 Tyr(705) signalling pathways. Br J Pharmacol. 2017;174:4140-4154.

[30] Yu N, Shen A, Chu J, et al. Qingda granule inhibits angiotensin II induced VSMCs proliferation through MAPK and PI3K/AKT pathways. J Ethnopharmacol. 2020;258:112767.

[31] Wang R, Sun Y, Yu W, et al. Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT. J Exp Clin Cancer Res. 2019;38:20.

[32] Xu H, Fang B, Du S, et al. Endothelial cell prostaglandin E2 receptor EP4 is essential for blood pressure homeostasis. JCI Insight. 2020;5:e138505.

[33] Lee JJ, Zhang WY, Yi H, et al. Anti-proliferative actions of 2-decylamino-5, 8-dimethoxy-1, 4-naphthoquinone in vascular smooth muscle cells. Biochem Biophys Res Commun. 2011;411:213-218.

[34] Yun W, Qian L, Cheng Y, Tao W, Yuan R, Xu H. Periplocymarin plays an efficacious Cardiotonic role via promoting calcium influx. Front Pharmacol. 2020;11:1292.

[35] Khan N, Jajeh F, Khan MI, Mukhtar E, Shabana SM, Mukhtar H. Sestrin-3 modulation is essential for therapeutic efficacy of cucurbitacin B in lung cancer cells. Carcinogenesis. 2017;38:184-195.

[36] Goel SA, Guo LW, Liu B, Kent KC. Mechanisms of post-intervention arterial remodelling. Cardiovasc Res. 2012;96:363-371.

[37] Heo KS, Fujiwara K, Abe J. Shear stress and atherosclerosis. Mol Cells. 2014;37:435-440.

[38] Liu M, Yan Q, Peng B, et al. Use of cucurbitacins for lung cancer research and therapy. Cancer Chemother Pharmacol. 2021;88:1-14.

[39] Ríos JL, Andújar I, Escandell JM, Giner RM, Recio MC. Cucurbitacins as inducers of cell death and a rich source of potential anticancer compounds. Curr Pharm Des. 2012;18:1663-1676.

[40] Ahmed MS, Halaweish FT. Cucurbitacins: potential candidates targeting mitogen-activated protein kinase pathway for treatment of melanoma. J Enzyme Inhib Med Chem. 2014;29:162-167.

[41] Roskoski R Jr. Cyclin-dependent protein serine/threonine kinase inhibitors as anticancer drugs. Pharmacol Res. 2019;139:471-488.

[42] Zheng Q, Liu Y, Liu W, et al. Cucurbitacin B inhibits growth and induces apoptosis through the JAK2/STAT3 and MAPK pathways in SH-SY5Y human neuroblastoma cells. Mol Med Rep. 2014;10:89-94.

[43] Chan KT, Meng FY, Li Q, et al. Cucurbitacin B induces apoptosis and S phase cell cycle arrest in BEL-7402 human hepatocellular carcinoma cells and is effective via oral administration. Cancer Lett. 2010;294:118-124.

[44] Kim SR, Seo HS, Choi HS, et al. Trichosanthes kirilowii ethanol extract and cucurbitacin D inhibit cell growth and induce apoptosis through inhibition of STAT3 activity in breast cancer cells. Evid Based Complement Alternat Med. 2013;2013:975350.

[45] Kim HJ, Park JH, Kim JK. Cucurbitacin-I, a natural cell-permeable triterpenoid isolated from Cucurbitaceae, exerts potent anticancer effect in colon cancer. Chem Biol Interact. 2014;219:1-8.

[46] Bicknell KA, Surry EL, Brooks G. Targeting the cell cycle machinery for the treatment of cardiovascular disease. J Pharm Pharmacol. 2003;55:571-591.

RIGHTS & PERMISSIONS

2024 The Author(s). Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.