Abstract
Objective
To investigate the effect and possible mechanism of hydroxysafflor yellow A (HSYA) on human immortalized keratinocyte cell proliferation and migration.
Methods
HaCaT cells were treated with HSYA. Cell proliferation was detected by the cell counting kit-8 assay, and cell migration was measured using wound healing assay and Transwell migration assay. The mRNA and protein expression levels of heparin-binding epidermal growth factor (EGF)-like growth factor (HBEGF), EGF receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR), and hypoxia-inducible factor-1α (HIF-1α) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Circ_0084443-overexpressing HaCaT cells and empty plasmid HaCaT cells were constructed using the lentiviral stable transfection and treated with HSYA. The expression of circ_0084443 was detected by qRT-PCR.
Results
HSYA (800 µmol/L) significantly promoted HaCaT cell proliferation and migration (P<0.05 or P<0.01). It also increased the mRNA and protein expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and increased the phosphorylation levels of PI3K and AKT (P<0.05 or P<0.01). Furthermore, HSYA promoted HaCaT cell proliferation and migration via the HBEGF/EGFR and PI3K/AKT/mTOR signaling pathways (P<0.01). Circ_0084443 attenuated the mRNA expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α (P<0.05). HSYA inhibited the circ_0084443 expression, further antagonized the inhibition of circ_0084443 on HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and promoted the proliferation of circ_0084443-overexpressing HaCaT cells (P<0.05 or P<0.01). However, HSYA could not influence the inhibitory effect of circ_0084443 on HaCaT cell migration (P>0.05).
Conclusion
HSYA played an accelerative role in HaCaT cell proliferation and migration, which may be attributable to activating HBEGF/EGFR and PI3K/AKT signaling pathways, and had a particular inhibitory effect on the keratinocyte negative regulator circ_0084443.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Rodrigues M, Kosaric N, Bonham CA, Gurtner GC. Wound healing: a cellular perspective. Physiol Rev 2019;99:665–706.
Dong Y, Yang Q, Sun X. Comprehensive analysis of cell therapy on chronic skin wound healing: a meta-analysis. Hum Gene Ther 2021;32:787–795.
Piipponen M, Li D, Landén NX. The immune functions of keratinocytes in skin wound healing. Int J Mol Sci 2020;21:8790.
Wang AX, Toma MA, Ma JX, Li DQ, Vij M, Chu TB, et al. Circular RNA hsa_circ_0084443 is upregulated in diabetic foot ulcer and modulates keratinocyte migration and proliferation. Adv Wound Care (New Rochelle) 2020;9:145–160.
Zhong H, Qian J, Xiao Z, Chen Y, He X, Sun C, et al. MicroRNA-133b inhibition restores EGFR expression and accelerates diabetes-impaired wound healing. Oxid Med Cell Longev 2021;2021:1–14.
Jere SW, Houreld NN, Abrahamse H. Role of the PI3K/AKT (mTOR and GSK3 β) signaling pathway and photobiomodulation in diabetic wound healing. Cytokine Growth Factor Rev 2019;50:52–59.
Fitsialos G, Bourget I, Augier S, Ginouvès A, Rezzonico R, Odorisio T, et al. HIF1 transcription factor regulates laminin-332 expression and keratinocyte migration. J Cell Sci 2008;121:2992–3001.
Zhou X, Guo Y, Yang K, Liu P, Wang J. The signaling pathways of traditional Chinese medicine in promoting diabetic wound healing. J Ethnopharmacol 2022;282:114662.
Ao H, Feng W, Peng C. Hydroxysafflor yellow A: a promising therapeutic agent for a broad spectrum of diseases. Evid Based Complement Alternat Med 2018;2018:1–17.
Gao SQ, Chang C, Niu XQ, Li LJ, Zhang Y, Gao JQ. Topical application of hydroxysafflor yellow A accelerates the wound healing in streptozotocin induced T1DM rats. Eur J Pharmacol 2018;823:72–78.
Gao SQ, Chang C, Li JJ, Li Y, Niu XQ, Zhang DP, et al. Co-delivery of deferoxamine and hydroxysafflor yellow A to accelerate diabetic wound healing via enhanced angiogenesis. Drug Deliv 2018;25:1779–1789.
Yang CC, Chang KW. Eicosanoids and HB-EGF/EGFR in cancer. Cancer Metastasis Rev 2018;37:385–395.
Yu D, Li X, Wang Z, Jiang S, Yan T, Fang K, et al. Role of AUF1 in modulating the proliferation, migration, and senescence of skin cells. Exp Ther Med 2021;23:45.
Zhi S, Li J, Kong X, Xie X, Zhang Q, Fang G. Insulin-like growth factor 2 mRNA binding protein 2 regulates proliferation, migration, and angiogenesis of keratinocytes by modulating heparanase stability. Bioengineered 2021;12:11267–11276.
Lu C, Kolbenschlag J, Nüssler AK, Ehnert S, McCaig CD, Cebron U, et al. Direct current electrical fields improve experimental wound healing by activation of cytokine secretion and Erk1/2 pathway stimulation. Life (Basel) 2021;11:1195.
Choi SH, Won KJ, Lee R, Cho HS, Hwang SH, Nah SY. Wound healing effect of gintonin involves lysophosphatidic acid receptor/vascular endothelial growth factor signaling pathway in keratinocytes. Int J Mol Sci 2021;22:10155.
Zheng T, Shao W, Tian J. Exosomes derived from ADSCs containing miR-378 promotes wound healing by targeting caspase-3. J Biochem Mol Toxicol 2021;35:e22881.
Liu Y, Xiong W, Wang CW, Shi JP, Shi ZQ, Zhou JD. Resveratrol promotes skin wound healing by regulating the miR-212/CASP8 axis. Lab Invest 2021;101:1363–1370.
Wu X, Xiao Y, Ma J, Wang A. Circular RNA: a novel potential biomarker for skin diseases. Pharmacol Res 2020;158:104841.
Stoll SW, Rittié L, Johnson JL, Elder JT. Heparin-binding EGF-like growth factor promotes epithelial-mesenchymal transition in human keratinocytes. J Invest Dermatol 2012;132:2148–2157.
Liu Y, Beyer A, Aebersold R. On the dependency of cellular protein levels on mRNA abundance. Cell 2016;165:535–550.
Correia de Sousa M, Gjorgjieva M, Dolicka D, Sobolewski C, Foti M. Deciphering miRNAs’ action through miRNA editing. Int J Mol Sci 2019;20:6249.
He Z, Xu X. Circ_0084443 inhibits wound healing via repressing keratinocyte migration through targeting the miR-17-3p/FOXO4 axis. Biochem Genet 2022;60:1236–1252.
Wang Q, Cang Z, Shen L, Peng W, Xi L, Jiang X, et al. Circ_0037128/miR-17-3p/AKT3 axis promotes the development of diabetic nephropathy. Gene 2021;765:145076.
Roudkenar MH, Fukumoto M, Roushandeh AM, Kuwahra Y, Uroshihara Y, Harada H, et al. Disturbance in the regulation of miR 17–92 cluster on HIF-1-α expression contributes to clinically relevant radioresistant cells: an in vitro study. Cytotechnology 2020;72:141–153.
Author information
Authors and Affiliations
Contributions
Zhang Y performed the experiments and wrote the manuscript. Wang AX designed the study, Xiao YW contributed the data analysis. Ma JX and Wang AX revised the manuscript. All authors contributed to the article and approved the final version.
Corresponding author
Ethics declarations
The authors declare no conflicts of interest.
Additional information
Supported by the Natural Science Fund of Liaoning Provincial Science and Technology Department (No. 20180530070), and Science and Technology Innovation Foundation of Dalian (No. 2020JJ27SN073)
Electronic Supplementary Material
Rights and permissions
About this article
Cite this article
Zhang, Y., Xiao, Yw., Ma, Jx. et al. Hydroxysafflor Yellow A Promotes HaCaT Cell Proliferation and Migration by Regulating HBEGF/EGFR and PI3K/AKT Pathways and Circ_0084443. Chin. J. Integr. Med. 30, 213–221 (2024). https://doi.org/10.1007/s11655-023-3607-2
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11655-023-3607-2