[1]芦周洲,高栋梁,王亚康.褪黑素调控GPX4/NF-κB p65信号通路干预铁死亡促进小鼠皮肤创面修复[J].陕西医学杂志,2025,54(2):181-186.[doi:DOI:10.3969/j.issn.1000-7377.2025.02.007]
　LU Zhouzhou,GAO Dongliang,WANG Yakang.Melatonin regulates the GPX4/NF-κB p65 signaling pathway to intervene ferroptosis and promote skin wound healing in mice[J].,2025,54(2):181-186.[doi:DOI:10.3969/j.issn.1000-7377.2025.02.007]
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褪黑素调控GPX4/NF-κB p65信号通路干预铁死亡促进小鼠皮肤创面修复
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
54
期数:
2025年2期
页码:
181-186
栏目:
基础研究
出版日期:
2025-02-05

文章信息/Info

Title:
Melatonin regulates the GPX4/NF-κB p65 signaling pathway to intervene ferroptosis and promote skin wound healing in mice
作者:
芦周洲1; 高栋梁2; 王亚康3
(1.宿州市第一人民医院烧伤整形科,安徽 宿州 234000; 2.延安大学附属医院烧伤整形外科,陕西 延安 716000; 3.西安市红会医院髋关节外科,陕西 西安 710054)
Author(s):
LU Zhouzhou1; GAO Dongliang2; WANG Yakang3
(1.Department of Burn and Plastic Surgery,Suzhou First People's Hospital, Suzhou 234000,China; 2.Department of Burn and Plastic Surgery,Yan'an University Affiliated Hospital,Yan'an 716000,China; 3.Department of Hip Surgery,Xi'an Honghui Hospital, Xi'an 710054,China)
关键词:
创面修复;  褪黑素;  铁死亡;  谷胱甘肽过氧化物酶4;  核因子-κB p65;  小鼠
Keywords:
Wound healing;  Melatonin;  Ferroptosis;  GPX4;  NF-κB p65;  Mice
分类号:
R -33
DOI:
DOI:10.3969/j.issn.1000-7377.2025.02.007
文献标志码:
A
摘要:
目的:探讨褪黑素对人皮肤成纤维细胞(HSF)增殖和铁死亡的作用,并探究其促进小鼠皮肤创面修复的分子机制。方法:采用CCK-8法检测HSF细胞增殖能力,选取细胞增殖率最高的褪黑素浓度进行后续实验。将HSF细胞随机分为对照组、谷氨酸组、谷氨酸+褪黑素组。谷氨酸+褪黑素组用100 μmol/L褪黑素预处理细胞60 min,随后除对照组外,另两组加入谷氨酸(10 mmol/L)诱导铁死亡。采用相应试剂盒检测HSF细胞活性氧(ROS)、谷胱甘肽(GSH)、丙二醛(MDA)、Fe2+水平。构建小鼠为全层皮肤损伤模型,并随机分为对照组和褪黑素组,每组10只。褪黑素组在皮肤损伤处外敷含5 mg褪黑素的凡士林乳膏,对照组外敷等量凡士林乳膏,每日清洁换药,连续9 d。计算术后第5、7、9天各组创面愈合率。采用HE和Masson染色观察第5、9天小鼠创面组织病理学变化及胶原纤维生成情况。采用免疫荧光染色法检测术后第9天创面组织中细胞增殖情况,Western blot检测术后第9天创面组织中谷胱甘肽过氧化物酶4(GPX4)、核因子(NF)-κB p65、p-NF-κB p65蛋白表达。结果:褪黑素浓度为100 μmol/L时HSF细胞增殖率最高,故选取100 μmol/L褪黑素进行后续实验。与对照组比较,谷氨酸组HSF细胞GSH水平降低,ROS、MDA、Fe2+水平升高(均P<0.05)。与谷氨酸组比较,谷氨酸+褪黑素组HSF细胞GSH水平升高,ROS、MDA、Fe2+水平降低(均P<0.05)。与对照组比较,褪黑素组小鼠术后第5、7、9天创面愈合率增加(均P<0.05)。与对照组比较,褪黑素组小鼠术后第5、9天创面组织再上皮化程度更高,胶原纤维生成比例增加(均P<0.05)。与对照组比较,褪黑素组小鼠术后第9天创面组织Ki-67免疫荧光强度增加(P<0.05)。与对照组比较,褪黑素组小鼠术后第9天创面组织GPX4蛋白表达水平升高,NF-κB p65、p-NF-κB p65蛋白表达水平降低(均P<0.05)。结论:褪黑素能够促进皮肤成纤维细胞增殖,抑制细胞铁死亡,促使小鼠创面组织再上皮化和胶原纤维形成,加快创面愈合,其机制可能与调控GPX4/NF-κB p65信号通路有关。
Abstract:
Objective:To investigate the effects of melatonin on the proliferation and ferroptosis of human skin fibroblasts(HSF)and to explore its molecular mechanisms in promoting skin wound healing in mice.Methods:The proliferation ability of HSF cells was detected using the CCK-8 assay,and the concentration of melatonin with the highest cell proliferation rate was selected for subsequent experiments.HSF cells were randomly divided into a control group,glutamate group,and glutamate+melatonin group.The glutamate+melatonin group was pretreated with 100 μmol/L melatonin for 60 minutes,and then glutamate(10 mmol/L)was added to induce ferroptosis in both the glutamate and glutamate+melatonin groups,except for the control group.The levels of reactive oxygen species(ROS),glutathione(GSH),malondialdehyde(MDA),and Fe2+ in HSF cells were detected using corresponding assay kits.A full-thickness skin injury model was constructed in mice,which were randomly divided into a control group and a melatonin group,with 10 mice in each group.The melatonin group applied a vaseline ointment containing 5 mg of melatonin to the skin injury site,while the control group applied an equal amount of vaseline ointment.The wounds were cleaned and dressed daily for 9 consecutive days.The wound healing rate was calculated on postoperative days 5,7 and 9.HE and Masson staining were used to observe the pathological changes and collagen fiber formation in the wound tissue on days 5 and 9.Immunofluorescence staining was used to detect cell proliferation in the wound tissue on day 9,and Western blot was used to detect the expression of glutathione peroxidase 4(GPX4),nuclear factor(NF)-κB p65,and p-NF-κB p65 proteins in the wound tissue on day 9.Results:When the concentration of melatonin was 100 μmol/L,the proliferation rate of HSF cells was the highest,so 100 μmol/L melatonin was selected for subsequent experiments.Compared with the control group,the GSH level in the glutamate group decreased,while the levels of ROS,MDA,and Fe2+ increased(all P<0.05).Compared with the glutamate group,the GSH level in the glutamate+melatonin group increased,while the levels of ROS,MDA and Fe2+ decreased(all P<0.05).Compared with the control group,the wound healing rate in the melatonin group increased on postoperative days 5,7 and 9(all P<0.05).Compared with the control group,the degree of re-epithelialization and the proportion of collagen fiber formation in the wound tissue of the melatonin group were higher on postoperative days 5 and 9(all P<0.05).Compared with the control group,the immunofluorescence intensity of Ki-67 in the wound tissue of the melatonin group on day 9 increased(P<0.05).Compared with the control group,the expression level of GPX4 protein in the wound tissue of the melatonin group on day 9 increased,while the expression levels of NF-κB p65 and p-NF-κB p65 proteins decreased(all P<0.05).Conclusion:Melatonin can promote the proliferation of skin fibroblasts,inhibit cell ferroptosis,promote re-epithelialization and collagen fiber formation in mouse wound tissue,accelerate wound healing,and its mechanism may be related to the regulation of the GPX4/NF-κB p65 signaling pathway.

参考文献/References:

[1] VASEY C,MCBRIDE J,PENTA K.Circadian rhythm dysregulation and restoration:The role of melatonin[J].Nutrients,2021,13(10):3480.
[2] 石艳红,邵英.生物钟相关基因CLOCK及褪黑素受体MT1和MT2基因与2型糖尿病患者肠道菌群及炎症因子相关性分析[J].陕西医学杂志,2023,52(3):335-339.
[3] BOCHEVA G,SLOMINSKI R M,JANJETOVIC Z,et al.Protective role of melatonin and its metabolites in skin aging[J].Int J Mol Sci,2022,23(3):1238.
[4] LIU W,YU M,XIE D,et al.Melatonin-stimulated MSC-derived exosomes improve diabetic wound healing through regulating macrophage M1 and M2 polarization by targeting the PTEN/AKT pathway[J].Stem Cell Res Ther,2020,11(1):259.
[5] DONG Y,CAO X,HUANG J,et al.Melatonin inhibits fibroblast cell functions and hypertrophic scar formation by enhancing autophagy through the MT2 receptor-inhibited PI3K/Akt/mTOR signaling[J].Biochim Biophys Acta Mol Basis Dis,2024,1870(1):166887.
[6] JING Y,BAI F,CHEN H,et al.Meliorating microcirculatory with melatonin in rat model of spinal cord injury using laser Doppler flowmetry[J].Neuroreport,2016,27(17):1248-1255.
[7] 刘湘慧,斯韬,梁婷,等.铁死亡发生机制及其在肝细胞癌中的作用研究进展[J].陕西医学杂志,2023,52(3):354-357.
[8] LIANG D,FENG Y,ZANDKARIMI F,et al.Ferroptosis surveillance independent of GPX4 and differentially regulated by sex hormones[J].Cell,2023,186(13):2748-2764.
[9] CUI S,LIU X,LIU Y,et al.Autophagosomes defeat ferroptosis by decreasing generation and increasing discharge of free Fe2+ in skin repair cells to accelerate diabetic wound healing[J].Adv Sci(Weinh),2023,10(25):e2300414.
[10] LUO L,DENG L,CHEN Y,et al.Identification of lipocalin 2 as a ferroptosis-related key gene associated with hypoxic-ischemic brain damage via STAT3/NF-κB signaling pathway[J].Antioxidants(Basel),2023,12(1):186.
[11] BOISMAL F,SERROR K,DOBOS G,et al.Skin aging:Pathophysiology and innovative therapies[J].Med Sci(Paris),2020,36(12):1163-1172.
[12] 刘荣奇,张会娜,范斌,等.凉血五花汤合舒肝丸加减联合Nd:YAG激光对黄褐斑患者皮肤屏障功能的影响[J].陕西中医,2023,44(5):608-611.
[13] 罗富强,余电柏,谢康麒,等.糖尿病足溃疡创面修复相关机制的研究进展[J].实用医学杂志,2023,39(2):158-163.
[14] TARGHAZEH N,REITER R J,RAHIMI M,et al.Oncostatic activities of melatonin:Roles in cell cycle,apoptosis,and autophagy[J].Biochimie,2022,202(3):34-48.
[15] RUSANOVA I,MARTÍNEZ-RUIZ L,FLORIDO J,et al.Protective effects of melatonin on the skin:Future perspectives[J].Int J Mol Sci,2019,20(19):4948.
[16] MARTO J,ASCENSO A,GONÇALVES L M,et al.Melatonin-based pickering emulsion for skin's photoprotection[J].Drug Deliv,2016,23(5):1594-607.
[17] TANG D,CHEN X,KANG R,et al.Ferroptosis:Molecular mechanisms and health implications[J].Cell Res,2021,31(2):107-125.
[18] URSINI F,MAIORINO M.Lipid peroxidation and ferroptosis:The role of GSH and GPx4[J].Free Radic Biol Med,2020,152(3):175-185.
[19] YANG W,WANG Y,ZHANG C,et al.Maresin1 protect against ferroptosis-induced liver injury through ROS inhibition and Nrf2/HO-1/GPX4 activation[J].Front Pharmacol,2022,13(3):865689.
[20] CHEN L,WANG X,LIU C,et al.Melatonin ameliorates atherosclerosis by suppressing S100a9-mediated vascular inflammation[J].Eur J Pharmacol,2023,957(3):175965.
[21] SENER G,SERT G,OZER SEHIRLI A,et al.Melatonin protects against pressure ulcer-induced oxidative injury of the skin and remote organs in rats[J].J Pineal Res,2006,40(3):280-7.
[22] 田馨如,贺佐分,岑丽君,等.MEBT/MEBO对慢性难愈合创面NF-κB p65、IκBα、IKK及其磷酸化蛋白表达水平的影响[J].中国烧伤创疡杂志,2022,34(3):153-161.
[23] 白治苗,刘玉锋,赵乐,等.核因子-κB p65和骨桥蛋白在子宫内膜癌组织与细胞中的表达及相关性研究[J].陕西医学杂志,2022,51(3):370-374.
[24] SHEN Y,TENG L,QU Y,et al.Anti-proliferation and anti-inflammation effects of corilagin in rheumatoid arthritis by downregulating NF-κB and MAPK signaling pathways[J].J Ethnopharmacol,2022,284(2):114791.
[25] QIANG L,YANG S,CUI Y H,et al.Keratinocyte autophagy enables the activation of keratinocytes and fibroblastsand facilitates wound healing[J].Autophagy,2021,17(9):2128-2143.
[26] CHEN C,LIN Z,LIU W,et al.Emodin accelerates diabetic wound healing by promoting anti-inflammatory macrophage polarization[J].Eur J Pharmacol,2022,936(1):175329.
[27] CHEN M,HE X,SUN Z,et al.Natural carrier-free self-assembled diterpene nanoparticles with its efficient anti-inflammation through the inhibition of NF-κB pathway for accelerated wound healing[J].Biomed Pharmacother,2023,165(4):115041.

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备注/Memo

备注/Memo:
[基金项目]陕西省自然科学基础研究计划项目(2019JQ-536)
更新日期/Last Update: 2025-02-04