Journal of Shandong University (Health Sciences) ›› 2026, Vol. 64 ›› Issue (1): 88-98.doi: 10.6040/j.issn.1671-7554.0.2025.0937

• Preclinical Medicine • Previous Articles     Next Articles

Mechanism of skin aging induced by cigarette smoke extract via the Klotho-CHRNA5 axis

WANG Yibo1, WANG Yingchao1, XU Linlin1, HUANG Shuhong2, DANG Ningning1   

  1. 1. Department of Dermatology, Shandong Provincial Hospital Affiliated to Shandong First Medical University;
    2. School of Clinical and Basic Medicine, Shandong First Medical University, Jinan 250021, Shandong, China
  • Published:2026-01-27

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Abstract: Objective To investigate how cigarette smoke accelerates skin aging by focusing on the interaction between the anti-aging protein Klotho and the tobacco-associated receptor CHRNA5, and explore potential targets for skin rejuvenation strategies. Methods HaCaT cells were treated with CSE(cigarette smoke extract). Cellular senescence and the expression of CHRNA5 and Klotho proteins were assessed by β-galactosidase staining and Western blotting. Subsequently, skin samples from mice aged 3-5, 6-7, and 14 months were analyzed using H&E staining, immunohistochemistry, Western blotting, and RT-qPCR to determine age-dependent changes in CHRNA5, Klotho and senescence-associated markers. Furthermore, in Klotho-knockdown mice, we assessed the efficiency of Klotho silencing and the effects of Klotho on both CHRNA5 expression and senescence markers using immunohistochemistry, Western blotting and RT-qPCR. Finally, HaCaT cells were transfected with siRNAs targeting Klotho or CHRNA5, and the expression of both genes and related senescence markers was measured by Western blotting and RT-qPCR to verify their regulatory relationship. Results CSE exposure induced senescence in HaCaT cells, as evidenced by a significant increase in β-galactosidase-positive cells(P=0.000 4), accompanied by the upregulation of CHRNA5(P=0.000 2)and the downregulation of Klotho(P=0.002 5). In 14-month-old mice, compared with 3-5-month-old controls, the epidermis was thinner(P<0.000 1), CHRNA5 was elevated(P=0.000 9), and Klotho was markedly decreased(P=0.000 1). In Klotho-knockdown mice, accelerated skin aging phenotypes were observed along with increased CHRNA5 expression(P=0.010 7). Similarly, Klotho-knockdown HaCaT cells exhibited enhanced senescence markers and elevated CHRNA5 expression(P=0.034 7). In contrast, CHRNA5 knockdown significantly reduced the expression of senescence-associated markers p16 and p21 in HaCaT cells(P=0.002 3; P=0.005 4). Conclusion CSE downregulates Klotho and upregulates CHRNA5, thereby promoting keratinocyte senescence andaccelerating skin aging.

Key words: Cigarette smoke extract, Klotho, CHRNA5, HaCaT, Skin aging

CLC Number: 

  • R574
[1] Furman D, Auwerx J, Bulteau AL, et al. Skin health and biological aging[J]. Nat Aging, 2025, 5(7): 1195-1206.
[2] 刘贺磊, 都日娜, 董俊娟, 等. 光老化与自噬机制的研究进展[J]. 内蒙古医学杂志, 2024, 56(3): 322-325.
[3] 付雨, 姜伟, 朱高培, 等. 基于多水平模型的山东省老年人健康相关生命质量及其影响因素[J]. 山东大学学报(医学版), 2025, 63(9): 57-64. FU Yu, JIANG Wei, ZHU Gaopei, et al. Health-related quality of life and its influencing factors of the elderly in Shandong Province based on multilevel models[J]. Journal of Shandong University(Health Science), 2025, 63(9): 57-64.
[4] Neves Cruz J, Santana de Oliveira M, Gomes Silva S, et al. Insight into the interaction mechanism of nicotine, NNK, and NNN with cytochrome P450 2A13 based on molecular dynamics simulation[J]. J Chem Inf Model, 2020, 60(2): 766-776.
[5] Böhm M, Stegemann A, Paus R, et al. Endocrine controls of skin aging[J]. Endocr Rev, 2025, 46(3): 349-375.
[6] Krutmann J, Bouloc A, Sore G, et al. The skin aging exposome[J]. J Dermatol Sci, 2017, 85(3): 152-161.
[7] Kuro-o M, Matsumura Y, Aizawa H, et al. Mutation of the mouse klotho gene leads to a syndrome resembling ageing[J]. Nature, 1997, 390(6655): 45-51.
[8] Espuch-Oliver A, Vázquez-Lorente H, Jurado-Fasoli L, et al. References values of soluble α-klotho serum levels using an enzyme-linked immunosorbent assay in healthy adults aged 18-85 years[J]. J Clin Med, 2022, 11(9): 2415. doi:10.3390/jcm11092415
[9] 任颖洁, 田庆华, 姜亦欣, 等. Koltho与脑血管疾病的关系及其相关生物学机制[J]. 中风与神经疾病杂志, 2023, 40(8): 763-768. REN Yingjie, TIAN Qinghua, JIANG Yixin, et al. Association of Klotho with cerebrovascular disease and asso-ciated biological mechanisms[J]. Journal of Apoplexy and Nervous Diseases, 2023, 40(8): 763-768.
[10] Kuro-O M. The Klotho proteins in health and disease[J]. Nat Rev Nephrol, 2019, 15(1): 27-44.
[11] Yamashita K, Yotsuyanagi T, Yamauchi M, et al. Klotho mice: a novel wound model of aged skin[J]. Plast Reconstr Surg Glob Open, 2014, 2(1): e101. doi:10.1097/GOX.0000000000000045
[12] Jin XH, Pi LQ, Lee WS. Expression pattern and role of klotho in human hair follicles[J]. Ann Dermatol, 2019, 31(5): 511-517.
[13] Pechlivanidou M, Ninou E, Karagiorgou K, et al. Autoimmunity to neuronal nicotinic acetylcholine receptors[J]. Pharmacol Res, 2023, 192: 106790. doi:10.1016/j.phrs.2023.106790
[14] 蔡佳莹, 王靖婷, 王增萍, 等. α5-nAChR对慢性应激肺腺癌荷瘤小鼠疑核c-Fos表达的影响[J]. 山东大学学报(医学版), 2025, 63(4): 69-74. CAI Jiaying, WANG Jingting, WANG Zengping, et al. Effect of α5-nAChR on the expression of c-Fos in nucleus ambiguus in chronic stress-induced lung adenocarcinoma mice[J]. Journal of Shandong University(Health Science), 2025, 63(4): 69-74.
[15] Wang J, Li XY, Zhang P, et al. CHRNA5 is over-expressed in patients with psoriasis and promotes psoriasis-like inflammation in mouse models[J]. J Investig Dermatol, 2022, 142(11): 2978-2987.
[16] Su Y, Han W, Giraldo C, et al. Effect of cigarette smoke extract on nitric oxide synthase in pulmonary artery endothelial cells[J]. Am J Respir Cell Mol Biol, 1998, 19(5): 819-825.
[17] Colucci M, Sarill M, Maddalena M, et al. Senescence in cancer[J]. Cancer Cell, 2025, 43(7): 1204-1226.
[18] OReilly S, Tsou PS, Varga J. Senescence and tissue fibrosis: opportunities for therapeutic targeting[J]. Trends Mol Med, 2024, 30(12): 1113-1125.
[19] Rahman A, Rehmani R, Pirvu DG, et al. Unlocking the therapeutic potential of marine collagen: a scientific exploration for delaying skin aging[J]. Mar Drugs, 2024, 22(4): 159. doi:10.3390/md22040159
[20] Li Z, Li J, Li L, et al. Klotho enhances stability of chronic kidney disease atherosclerotic plaques by inhibiting GRK2/PLC-β-mediated endoplasmic reticulum stress in macrophages via modulation of the ROS/SHP1 pathway[J]. Sci Rep, 2024, 14(1): 32091. doi:10.1038/s41598-024-83596-w
[21] Awasthi R, Manger PT, Khare RK, et al. Klotho protein: a new insight into the pathogenesis of essential hypertension[J]. Clin Hypertens, 2024, 30(1): 36. doi:10.1186/s40885-024-00294-5
[22] Chen P, Tang YF, Luo L, et al. Lower serum Klotho level and higher systemic immune-inflammation index: an inverse correlation[J]. BMC Geriatr, 2023, 23(1): 650. doi:10.1186/s12877-023-04349-4
[23] Jin MH, Lv PF, Chen GY, et al. Klotho ameliorates cyclosporine A-induced nephropathy via PDLIM2/NF-kB p65 signaling pathway[J]. Biochem Biophys Res Commun, 2017, 486(2): 451-457.
[24] Yan F, Zhao Q, Li Y, et al. The role of oxidative stress in ovarian aging: a review[J]. J Ovarian Res, 2022, 15(1): 100. doi:10.1186/s13048-022-01032-x
[25] Zhang Y, Jia YF, Li P, et al. Reciprocal activation of α5-nAChR and STAT3 in nicotine-induced human lung cancer cell proliferation[J]. J Genet Genomics, 2017, 44(7): 355-362.
[26] Ernster VL, Grady D, Miike R, et al. Facial wrinkling in men and women, by smoking status[J]. Am J Public Health, 1995, 85(1): 78-82.
[27] Morita A. Tobacco smoke causes premature skin aging[J]. J Dermatol Sci, 2007, 48(3): 169-175.
[28] Ogrodnik M, Carlos Acosta J, Adams PD, et al. Guidelines for minimal information on cellular senescence experimentation in vivo[J]. Cell, 2024, 187(16): 4150-4175.
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