Kirenol inhibits TNF-α-induced proliferation and migration of HaCaT cells by regulating NF-κB pathway

Main Article Content

Jin Li
Fang Ren
Wenliang Yan
Hong Sang

Keywords

kirenol, psoriasis, proliferation, migration, inflammation

Abstract

Psoriasis is a common chronic, inflammatory skin disease possessing properties of inflammatory cell infiltration and excessive proliferation of keratinocytes, the occurrence and development of which remain fully elucidated. Therefore, the study was designed to determine the effects of kirenol (50, 100 and 200 μg/mL) on Cultured Human Keratinocytes (cells) (HaCaT) in vitro and reveal its molecular mechanism. The in vitro psoriasis model was established utilizing tumor necrosis factor-α (TNF-α)-stimulated HaCaT cells. Kirenol, a diterpenoid compound, was applied at different concentrations (50, 100 and 200 μg/mL) to HaCaT cells for 24 h. The Cell Counting Kit-8 (CCK-8) and thymidine monobromodeoxyuridine (BrdU) assays were used to assess cell viability and proliferation, followed by assessment of cell migration by Transwell assay. Subsequently, inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA), and Western blot assay was used to evaluate expressions of p65, p-p65, IκBα and p-IκBα. Activities of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and malondialdehyde (MDA) contents were measured spectrophotometrically. The results demonstrated that TNF-α induced a significant increase in cell viability and inflammatory cytokines, including expressions of Interleukin (IL)-6, IL-8, IL-22 and IL-1β in HaCaT cells, which was dose-dependently inhibited by kirenol. Similarly, TNF-α-induced cell migration was also suppressed by kirenol treatment. Furthermore, TNF-α stimuli induced the upregulation of phosphorylation levels of p65 and IκBα as well as p-p65–p65 and p-IκBα–IκBα ratios, whereas kirenol significantly suppressed the activation of cellular nuclear factor-kappa B (NF-κB) signaling pathway. In addition, kirenol significantly decreased the level of MDA but increased the levels of SOD, CAT and GSH in a dose-dependent manner. These results proposed that kirenol could inhibit the proliferation, migration, expression of inflammatory factors, and oxidative stress in HaCaT cells via suppressing NF-κB signaling pathway.

Abstract 252 | PDF Downloads 96 HTML Downloads 62 XML Downloads 59

References

Allergologia et Immunopathologia., 2020. Allergologia et immunopathologia: A new journey. Editorial. Allergologia et Immunopathologia (Madr) 48: 521–522. 10.1016/j.aller.2020.10.001

Aquino, T.M., Calvarido, M.G. and North, J.P., 2021. Interleukin 36 expression in psoriasis variants and other dermatologic diseases with psoriasis-like histopathologic features. Journal of Cutaneous Pathology. Online ahead of print. 10.1111/cup.14115

Benhadou, F., Mintoff, D. and Del Marmol, V., 2019. Psoriasis: keratinocytes or immune cells—which is the trigger? Dermatology (Basel, Switzerland) 235(2): 91–100. 10.1159/000495291

Boehncke, W.-H. and Schön, M.P., 2015. Psoriasis. Lancet (London, England) 386: 983–994. 10.1016/S0140-6736(14)61909-7

Callis Duffin, K., Gottlieb, A.B., Merola, J.F., Latella, J., Garg, A. and Armstrong, A.W., 2017. Defining outcome measures for psoriasis: the IDEOM report from the GRAPPA 2016 annual meeting. Journal of Rheumatology 44: 701–702. 10.3899/jrheum.170151

Cho, J.-W., Lee, K.-S. and Kim, C.-W., 2007. Curcumin attenuates the expression of IL-1β, IL-6, and TNF-α as well as cyclin E in TNF-α-treated HaCaT cells; NF-κB and MAPKs as potential upstream targets, International Journal of Molecular Medicine 19: 469–474. PMid: 17273796

Choi, D.H. and Hwang, H.S., 2019. Anti-inflammation activity of Brazilin in TNF-α-induced human psoriasis dermatitis skin model. Applied Biological Chemistry 62: 1–9. 10.1186/s13765-019-0455-z

Cooper, K.D., 1990. Psoriasis. Leukocytes and cytokines. Dermatologic Clinics 8(4): 737–745. PMid: 2249364

Evans, E.A., Sayers, S.R., Kodji, X., Xia, Y., Shaikh, M., Rizvi, A., et al. 2020. Psoriatic skin inflammation induces a pre-diabetic phenotype via the endocrine actions of skin secretome. Molecular Metabolism 41: 101047. 10.1016/j.molmet.2020.101047

Griffiths, C.E. and Barker, J.N., 2007. Pathogenesis and clinical features of psoriasis. Lancet (London, England) 370: 263–271. 10.1016/S0140-6736(07)61128-3

Hayden, M.S. and Ghosh, S., 2011. NF-κB in immunobiology. Cell Research 21: 223–244. 10.1038/cr.2011.13

Hoegler, K.M., John, A.M., Handler, M.Z. and Schwartz, R.A., 2018. Generalized pustular psoriasis: a review and update on treatment. Journal of the European Academy of Dermatology and Venereology (JEADV) 32: 1645–1651. 10.1111/jdv.14949

Ibrahim, S.R.M., Altyar, A.E., Sindi, I.A., El-Agamy, D.S., Abdallah, H.M., Mohamed, S.G.A. and Mohamed, G.A., 2021. Kirenol: a promising bioactive metabolite from siegesbeckia species: a detailed review. Journal of Ethnopharmacology 5(281): 114552. 10.1016/j.jep.2021.114552

Kim, M.Y., Choi, Y.W. and Hwang, H.S., 2021. Regulatory effect on skin differentiation by mevastatin in psoriasis model using TNF-α and IL-17 induced HaCaT cells. Biotechnology and Bioprocess Engineering 26: 348–358. 10.1007/s12257-020-0368-z

Kim, T.-G., Kim, D.S., Kim, H.-P. and Lee, M.-G., 2014a. The pathophysiological role of dendritic cell subsets in psoriasis. BMB Reports 47: 60–68. 10.5483/bmbrep.2014.47.2.014

Kim, M.-B., Song, Y., Kim, C. and Hwang, J.-K., 2014b. Kirenol inhibits adipogenesis through activation of the Wnt/β-catenin signaling pathway in 3T3-L1 adipocytes. Biochemical and Biophysical Research Communications 445: 433–438. 10.1016/j.bbrc.2014.02.017

Langley, R.G.B., Krueger, G.G. and Griffiths, C.E.M., 2005. Psoriasis: epidemiology, clinical features, and quality of life. Annals of the Rheumatic Diseases 64(Suppl 2): ii18–23; discussion ii24–25. 10.1136/ard.2004.033217

Lawrence, T., 2009. The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harbor Perspectives in Biology 1: a001651. 10.1101/cshperspect.a001651

Lee, N., Chung, Y.C., Kang, C.I., Park, S.-M. and Hyun, C.-G., 2019. 7, 8-Dimethoxycoumarin attenuates the expression of IL-6, IL-8, and CCL2/MCP-1 in TNF-α-treated HaCaT cells by potentially targeting the NF-κB and MAPK pathways. Cosmetics 6(3): 41. 10.3390/cosmetics6030041

Lin, Z.-M., Ma, M., Li, H., Qi, Q., Liu, Y.-T., Yan, Y.-X., et al. 2018. Topical administration of reversible SAHH inhibitor ameliorates imiquimod-induced psoriasis-like skin lesions in mice via suppression of TNF-α/IFN-γ-induced inflammatory response in keratinocytes and T cell-derived IL-17. Pharmacological Research 129: 443–452. 10.1016/j.phrs.2017.11.012

Liu, W., Li, Y. and Li, C., 2020a. Kirenol exhibits the protective role against N-methyl-N-nitrosourea-induced gastric cancer in rats via modulating the oxidative stress and inflammatory markers. Journal of Environmental Pathology, Toxicology and Oncology: Official Organ of the International Society for Environmental Toxicology and Cancer 39: 345–355. 10.1615/JEnvironPatholToxicolOncol.2020035475

Liu, A., Zhao, W., Zhang, B., Tu, Y., Wang, Q. and Li, J., 2020b. Cimifugin ameliorates imiquimod-induced psoriasis by inhibiting oxidative stress and inflammation via NF-κB/MAPK pathway. Bioscience Reports 40(6): BSR20200471. 10.1042/BSR20200471

Lu, Y., Xiao, J., Wu, Z.-W., Wang, Z.-M., Hu, J., Fu, H.-Z., et al. 2012. Kirenol exerts a potent anti-arthritic effect in collagen-induced arthritis by modifying the T cells balance. Phytomedicine : International Journal of Phytotherapy and Phytopharmacology 19: 882–889. 10.1016/j.phymed.2012.04.010

Na Takuathung, M., Wongnoppavich, A., Pitchakarn, P., Panthong, A., Khonsung, P., Chiranthanut, N., Soonthornchareonnon, N. and Sireeratawong, S., 2017. Effects of wannachawee recipe with antipsoriatic activity on suppressing inflammatory cytokine production in hacat human keratinocytes. Evidence-Based Complementary and Alternative Medicine 2017: 5906539. 10.1155/2017/5906539.

Nair, R.P., Duffin, K.C., Helms, C., Ding, J., Stuart, P.E., Goldgar, D., et al. 2009. Genome-wide scan reveals association of psoriasis with IL-23 and NF-kappaB pathways. Nature Genetics 41: 199–204. 10.1038/ng.311

Nickoloff, B.J., 2006. Keratinocytes regain momentum as instigators of cutaneous inflammation. Trends in Molecular Medicine 12(3): 102–106. 10.1016/j.molmed.2006.01.001

Patel, M., Horgan, P.G., McMillan, D.C. and Edwards, J., 2018. NF-κB pathways in the development and progression of colorectal cancer. Translational Research : The Journal of Laboratory and Clinical Medicine 197: 43–56. 10.1016/j.trsl.2018.02.002

Ren, J., Yang, M., Chen, J., Ma, S. and Wang, N., 2020. Anti-inflammatory and wound healing potential of kirenol in diabetic rats through the suppression of inflammatory markers and matrix metalloproteinase expressions. Biomedicine & Pharmacotherapy 129: 110475. 10.1016/j.biopha.2020.110475

Srivastava, A.K., Chand Yadav, T., Khera, H.K., Mishra, P., Raghuwanshi, N., Pruthi, V. and Prasad, R., 2021. Insights into interplay of immunopathophysiological events and molecular mechanistic cascades in psoriasis and its associated comorbidities. Journal of Autoimmunity 118: 102614. 10.1016/j.jaut.2021.102614

Subhan, F., Kang, H.Y., Lim, Y., Ikram, M., Baek, S.-Y., Jin, S., et al. 2017. Fish scale collagen peptides protect against CoCl2/TNF-α-induced cytotoxicity and inflammation via inhibition of ROS, MAPK, and NF-κB pathways in HaCaT cells. Oxidative Medicine and Cellular Longevity 2017: 9703609. 10.1155/2017/9703609.

Wang, Q., Liang, Y.-Y., Li, K.-W., Li, Y., Niu, F.-J., Zhou, S.-J., et al. 2021a. Herba siegesbeckiae: a review on its traditional uses, chemical constituents, pharmacological activities and clinical studies. Journal of Ethnopharmacology 275: 114117. 10.1016/j.jep.2021.114117

Wang, A., Wei, J., Lu, C., Chen, H., Zhong, X., Lu, Y., et al. 2019. Genistein suppresses psoriasis-related inflammation through a STAT3-NF-κB-dependent mechanism in keratinocytes. International Immunopharmacology 69: 270–278. 10.1016/j.intimp.2019.01.054

Wang, Y., Xu, J., Alarifi, S. and Wang, H., 2021b. Kirenol inhibited the cell survival and induced apoptosis in human thyroid cancer cells by altering PI3K/AKT and MAP kinase signaling pathways. Environmental Toxicology 36: 811–820. 10.1002/tox.23083

Wang, J.-P., Zhou, Y.-M. and Zhang, Y.-H., 2012. Kirenol production in hairy root culture of siegesbeckea orientalis and its antimicrobial activity. Pharmacognosy Magazine 8: 149–155. 10.4103/0973-1296.96569

Wang, Z.-M., Zhu, S.-G., Wu, Z.-W., Lu, Y., Fu, H.-Z. and Qian, R.-Q., 2011. Kirenol upregulates nuclear annexin-1 which interacts with NF-κB to attenuate synovial inflammation of collagen-induced arthritis in rats. Journal of Ethnopharmacology 137: 774–782. 10.1016/j.jep.2011.06.037

Winchell, S.A. and Watts, R.A., 1988. Relaxation therapies in the treatment of psoriasis and possible pathophysiologic mechanisms. Journal of the American Academy of Dermatology 18: 101–104.

Wolk, K., Haugen, H.S., Xu, W., Witte, E., Waggie, K., Anderson, M., et al. 2009. IL-22 and IL-20 are key mediators of the epidermal alterations in psoriasis while IL-17 and IFN-γ are not. Journal of Molecular Medicine 87: 523–536.

Wu, J., Li, Q., Jin, L., Qu, Y., Liang, B.-B., Zhu, X.-T., et al. 2019. Kirenol Inhibits the function and inflammation of fibroblast-like synoviocytes in rheumatoid arthritis in vitro and in vivo. Frontiers in Immunology 10: 1304. 10.3389/fimmu.2019.01304

Xiuhong, L., Yajun, D.U., Guoxing, L., Guomei, D., Xin, T. and Juan, X., 2019. Kirenol relieves dextran sulfate sodium-induced ulcerative colitis in mice by inhibiting inflammatory cytokines and inducing CD4 T lymphocyte apoptosis. Nan Fang Yi Ke Da Xue Xue Bao (Journal of Southern Medical University) 39: 1387–1392. 10.12122/j.issn.1673-4254.2019.12.01

Xu, F., Xu, J., Xiong, X. and Deng, Y., 2019. Salidroside inhibits MAPK, NF-κB, and STAT3 pathways in psoriasis-associated oxidative stress via SIRT1 activation. Redox Report: Communications in Free Radical Research 24: 70–74. 10.1080/13510002.2019.1658377

Yang, H. and Zheng, J., 2020. Influence of stress on the development of psoriasis. Clinical and Experimental Dermatology 45: 284–288. 10.1111/ced.14105

Yi, W., Wen, Y., Tan, F., Liu, X., Lan, H., Ye, H. and Liu, B., 2019. Impact of NF-κB pathway on the apoptosis-inflammation-autophagy crosstalk in human degenerative nucleus pulposus cells. Aging 11: 7294–7306. 10.18632/aging.102266

Zhang, Q., Lenardo, M.J. and Baltimore, D., 2017. 30 Years of NF-κB: a blossoming of relevance to human pathobiology. Cell 168: 37–57. 10.1016/j.cell.2016.12.012

Zhou, S., Li, Q., Wu, H. and Lu, Q., 2020. The pathogenic role of innate lymphoid cells in autoimmune-related and inflammatory skin diseases. Cellular & Molecular Immunology 17: 335–346. 10.1038/s41423-020-0399-6