RSK2通过调控人皮肤成纤维细胞的增殖与迁移促进皮肤创面修复

doi:10.3969/j.issn.2095-9400.2020.07.001

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摘要目的：探究RSK2对皮肤创面修复的影响及其可能机制。方法：采用Western blot与免疫荧光染色实验检测皮肤创伤组织中RSK2的表达差异。通过人原代真皮成纤维细胞（HDF）划痕实验、CCK-8增殖实验及Western blot实验，在细胞水平探究RSK2抑制剂Bix 02565对HDF的作用。通过HE染色、Masson染色与免疫荧光染色，在体内水平观察RSK2抑制剂对皮肤创面修复的影响。结果：Western blot及免疫荧光实验结果显示皮肤创伤并不影响皮肤成纤维细胞中RSK2总蛋白的表达，而在体外水平上利用Bix 02565抑制RSK2的活性可显著抑制成纤维细胞的增殖与迁移，抑制G1/S-特异性周期蛋白-D1（Cyclin D1）与纤维粘连蛋白（Fibronectin）的表达，在动物水平RSK2抑制剂可抑制成纤维细胞的增殖细胞核抗原（PCNA）与胶原纤维生成，抑制血小板-内皮细胞黏附分子（CD31）的表达，阻滞血管的新生，延缓小鼠皮肤创面修复速率。结论：RSK2可通过促进成纤维细胞的增殖与迁移、血管新生，进而促进皮肤创面修复。

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	汪佳男
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	丛维涛

关键词 ： RSK2, Bix 02565, 成纤维细胞, 创面修复

Abstract：Objective: To investigate the effect of RSK2 on skin wound healing and its mechanism. Methods: Western blot and immunofluorescence staining were used to detect the expression of RSK2 under wound conditions. Scratch test, CCK-8 detection and Western blot were used to investigate the role of RSK2 inhibitor Bix 02565 on human primary dermal fibroblasts (HDF) at the cellular level. In vivo, HE staining, Masson staining and immunofluorescence staining were used to detect the effect of RSK2 inhibitor on skin wound repair. Results: Western blot and immunofluorescence staining showed that the skin trauma modal did not affect the total protein expression of RSK2 in fibroblasts, while the use of Bix 02565 to inhibit the activity of RSK2 in vitro could significantly inhibit the proliferation and migration of fibroblasts, and inhibit the expression of G1/S-specific cyclin-D1 and Fibronectin. In vivo, the inhibitor restrained proliferating cell nuclear antigen (PCNA) and collagen fibers production of fibroblasts, inhibited platelet-endothelial cell adhesion molecule (CD31), blocked vessels regeneration and delayed the repair rate of wound healing in mice. Conclusion: RSK2 facilitates skin wound healing mainly by promoting the proliferation and migration of fibroblasts and via angiogenesis.

Key words： RSK2 Bix 02565 fibroblast wound healing

基金资助:国家自然科学基金资助项目（81673077）。

通讯作者: 丛维涛，教授，硕士生导师，Email：cwt97126@126.com。

作者简介: 汪佳男（1994-），男，浙江宁波人，硕士生。

链接本文:

https://xb.wmu.edu.cn/CN/10.3969/j.issn.2095-9400.2020.07.001 或 https://xb.wmu.edu.cn/CN/Y2020/V50/I7/517

[1] 李小琼, 魏志平, 刘彦群. 皮肤屏障功能与皮肤病[J]. 中国麻风皮肤病杂志, 2014, 30(12): 731-733.
[2] LAGARES D, GHASSEMI-KAKROODI P, TREMBLAY C, et al. ADAM10-mediated ephrin-B2 shedding promotes myofibroblast activation and organ fibrosis[J]. Nat Med, 2017, 23(12): 1405-1415.
[3] RODRIGUES M, KOSARIC N, BONHAM C A, et al. Wound healing: A cellular perspective[J]. Physiol Rev, 2019, 99(1): 665-706.
[4] JINNIN M. Mechanisms of skin fibrosis in systemic sclerosis[J]. J Dermatol, 2010, 37(1): 11-25.
[5] CHO Y Y. RSK2 and its binding partners in cell proliferation, transformation and cancer development[J]. Arch Pharm Res, 2017, 40(3): 291-303.
[6] LIU K, CHO Y Y, YAO K, et al. Eriodictyol inhibits RSK2-ATF1 signaling and suppresses EGF-induced neoplastic cell transformation[J]. J Biol Chem, 2011, 286(3): 2057-2066.
[7] PENG C, CHO Y Y, ZHU F, et al. Phosphorylation of caspase-8 (Thr-263) by ribosomal S6 kinase 2 (RSK2) mediates caspase-8 ubiquitination and stability[J]. J Biol Chem, 2011, 286(9): 6946-6954.
[8] KIM J E, HEO Y S, LEE K W. Osajin inhibits solar UV-induced cyclooxygenase-2 expression through direct inhibition of RSK2[J]. J Cell Biochem, 2017, 118(11): 4080-4087.
[9] ZHU F, ZYKOVA T A, PENG C, et al. Phosphorylation of H2AX at Ser139 and a new phosphorylation site Ser16 by RSK2 decreases H2AX ubiquitination and inhibits cell transformation[J]. Cancer Res, 2011, 71(2): 393-403.
[10] ZHOU Y, YAMADA N, TANAKA T, et al. Crucial roles of RSK in cell motility by catalysing serine phosphorylation of EphA2[J]. Nat Commun, 2015, 6: 7679.
[11] LUDWIK K A, MCDONALD O G, BRENIN D R, et al. ERα-mediated nuclear sequestration of RSK2 is required for ER+ breast cancer tumorigenesis[J]. Cancer Res, 2018, 78(8): 2014-2025.
[12] 吴艳红, 邓安彦, 李晓红, 等. 人皮肤成纤维细胞的分离培养及鉴定[J]. 四川生理科学杂志, 2013, 35(4): 150-152.
[13] 王玲玲, 马峰, 张玉成, 等. 人成纤维细胞的体外分离、纯化培养及细胞鉴定[J]. 中国老年学杂志, 2012, 32(6): 1215-1216.
[14] 林蓓蓓, 郁引飞. 碱性成纤维细胞生长因子对大鼠II度烧伤伤口的促愈合作用[J]. 温州医科大学学报, 2017, 47(10): 723-729.
[15] LINEHAN J L, HARRISON O J, HAN S J, et al. Non-classical immunity controls microbiota impact on skin immunity and tissue repair[J]. Cell, 2018, 172(4): 784-796.
[16] EMING S A, WYNN T A, MARTIN P. Inflammation and metabolism in tissue repair and regeneration[J]. Science, 2017, 356(6342): 1026-1030.
[17] LYNCH M D, WATT F M. Fibroblast heterogeneity: implications for human disease[J]. J Clin Invest, 2018, 128(1): 26-35.
[18] DRISKELL R R, LICHTENBERGER B M, HOSTE E, et al. Distinct fibroblast lineages determine dermal architecture in skin development and repair[J]. Nature, 2013, 504(7479): 277-281.
[19] REINKE J M, SORG H. Wound repair and regeneration[J]. Eur Surg Res, 2012, 49(1): 35-43.
[20] RODERO M P, KHOSROTEHRANI K. Skin wound healing modulation by macrophages[J]. Int J Clin Exp Pathol, 2010, 3(7): 643-653.
[21] ZHANG L J, SEN G L, WARD N L, et al. Antimicrobial peptide LL37 and MAVS signaling drive interferon-β production by epidermal keratinocytes during skin injury[J]. Immunity, 2016, 45(1): 119-130.
[22] KEYES B E, LIU S, ASARE A, et al. Impaired epidermal to dendritic T cell signaling slows wound repair in aged skin [J]. Cell, 2016, 167(5): 1323-1338.
[23] ZHANG H, SU Y, WANG J, et al. Ginsenoside Rb1 promotes the growth of mink hair follicle via PI3K/AKT/GSK-3β signaling pathway[J]. Life Sci, 2019, 229: 210-218.
[24] SAITO Y, VANDENHEEDE J R, COHEN P. The mechanism by which epidermal growth factor inhibits glycogen synthase kinase 3 in A431 cells[J]. Biochem J, 1994, 303(Pt 1): 27-31.
[25] LEE C J, MOON S J, JEONG J H, et al. Kaempferol targeting on the fibroblast growth factor receptor 3-ribosomal S6 kinase 2 signaling axis prevents the development of rheumatoid arthritis[J]. Cell Death Dis, 2018, 9(3): 401.
[26] YOO S M, LEE C J, AN H J, et al. RSK2-mediated ELK3 activation enhances cell transformation and breast cancer cell growth by regulation of c-fos promoter activity[J]. Int J Mol Sci, 2019, 20(8): 1994.
[27] HAMAOKA Y, NEGISHI M, KATOH H. EphA2 is a key effector of the MEK/ERK/RSK pathway regulating glioblastoma cell proliferation[J]. Cell Signal, 2016, 28(8): 937-945.
[28] ISHIHARA J, ISHIHARA A, STARKE R D, et al. The heparin binding domain of von Willebrand factor binds to growth factors and promotes angiogenesis in wound healing[J]. Blood, 2019, 133(24):2559-2569.
[29] CHOI J K, JANG J H, JANG W H, et al. The effect of epidermal growth factor (EGF) conjugated with low-molecular-weight protamine (LMWP) on wound healing of the skin[J]. Biomaterials, 2012, 33(33): 8579-8590.