基于AMPK/p38MAPK/NF-κB通路探究湿润烧伤膏改善咪喹莫特诱导小鼠银屑病样的机制研究

摘要/Abstract

摘要： 目的：采用咪喹莫特（imiquimod， IMQ）诱导的银屑病样小鼠模型和白细胞介素-17A（interleukin-17A， IL-17A）诱导的人角质形成细胞（HaCaT）银屑病样模型，研究湿润烧伤膏（moist exposed burn ointment， MEBO）对银屑病的治疗作用和干预机制。方法：体内实验采用IMQ诱导BALB/c小鼠建立银屑病样模型，连续6 d MEBO干预后，评估小鼠皮损情况（PASI评分）和脾脏指数，H&E染色观察皮肤组织病理学变化，ELISA检测皮肤中银屑病相关细胞因子，Western blot检测AMPK/p38MAPK/NF-κB通路蛋白的表达。体外实验利用IL-17A诱导HaCaT细胞模拟银屑病样角质形成细胞模型，MTS评估MEBO对细胞增殖的影响，ELISA检测IL-6和IL-1β的表达水平，Western blot检测AMPK/p38MAPK/NF-κB蛋白的表达。结果：与模型组比较，MEBO干预显著改善小鼠银屑病样皮损症状，降低PASI评分和脾脏指数，减轻皮损病理损伤，降低皮损中银屑病相关细胞因子的水平，下调p-NF-κB和p-p38MAPK蛋白表达水平，上调p-AMPK蛋白表达水平；体外细胞模型中，与模型组比较，MEBO能够抑制细胞增殖，降低IL-6、IL-1β表达及p-p38MAPK、p-NF-κB蛋白表达水平，激活p-AMPK蛋白表达。结论：湿润烧伤膏通过p38MAPK/NF-κB/AMPK通路抑制IL-23/Th17轴介导的炎症反应，降低炎性细胞因子水平，改善小鼠银屑病样症状。

关键词: font-size:medium, ">银屑病；湿润烧伤膏；咪喹莫特；IL-17A；IL-23/Th17

Abstract: Objective： To investigate the therapeutic effect and intervention mechanism of Moist Exposed Burn Ointment (MEBO) on psoriasis， imiquimod (IMQ)-induced psoriasis-like mouse model and interleukin-17A (IL-17A)-induced psoriasis-like model of human keratinocyte-forming cells (HaCaT) were used. Methods： In vivo experiments were performed using IMQ-induced BALB/c mice to establish a psoriasis-like mouse model. After 6 consecutive days of MEBO intervention， mice were evaluated for skin lesions (psoriasis lesion area and disease severity scores) and splenic index. Hematoxylin-eosin staining was used to observe the histopathologic changes in the skin. Enzyme-linked immunosorbent assay was used to detect psoriasis-related cytokines in the skin.Western blot was used to detect the expression of AMPK/p38MAPK/NF-κB pathway proteins. For in vitro experiments， IL-17A was utilized to induce HaCaT cells to mimic a psoriasis-like keratinocyte model.MTS was used to assess the effect of MEBO on cell proliferation. The expression levels of IL-6 and IL-1β were detected by enzyme-linked immunosorbent assay.Western blot was used to detect the expression of AMPK/p38MAPK/NF-κB proteins. Results： Compared with the model group， MEBO intervention significantly improved the symptoms of psoriasis-like skin lesions， reduced PASI score and spleen index， and attenuated the pathological damage of skin lesions in mice.MEBO reduced the levels of psoriasis-related cytokines in skin lesions， down-regulated the levels of p-NF-κB and p-p38MAPK protein expression， and up-regulated the levels of p-AMPK protein expression. In the in vitro cell model， compared with the model group， MEBO could inhibit cell proliferation and reduce IL-6 and IL-1β expression. At the same time， MEBO could reduce p-p38MAPK and p-NF-κB protein expression levels and activate p-AMPK protein expression. Conclusion： MEBO can inhibit the suppression of IL-23/Th17 axis-mediated inflammatory response through the p38MAPK/NF-κB/AMPK pathway， reduce the level of inflammatory cytokines， and ameliorate psoriasis-like symptoms in mice.

Key words: font-size:medium, ">Psoriasis； Moist exposed burn ointment； Cytokines； IL-17A； IL-23/Th17

中图分类号:

门钟兰1, 黄竹芸2, 章彦文3, 褚付奥1, 孙语迪2, 孙双勇1, 2, 3. 基于AMPK/p38MAPK/NF-κB通路探究湿润烧伤膏改善咪喹莫特诱导小鼠银屑病样的机制研究[J]. 中国药物评价, 2025, 42(1): 21-27.

MEN Zhonglan1, HUANG Zhuyun2, ZHANG Yanwen3, CHU Fuao1, SUN Yudi2, SUN Shuangyong1, 2, 3. Mechanism of Moist Exposed Burn Ointment on Imiquimote-Induced Psoriasis in Mice through AMPK/p38MAPK/NF-κB Pathway[J]. CHINESE JOURNAL OF DRUG EVALUATION, 2025, 42(1): 21-27.

参考文献

[1]John SM， Loring B， Michalek IM. A systematic review of worldwide epidemiology of psoriasis[J]. J Eur Acad Dermatol Venereol， 2017， 31(2)： 205-212.
     [2]Boehncke WH， Schn MP. Psoriasis[J]. Lancet， 2015， 386(9997)： 983-994.
     [3]Rendon A， Schkel K. Psoriasis Pathogenesis and Treatment[J]. Int J Mol Sci， 2019， 20(6)： 1475.
     [4]Zhang L， Ma X， Shi R， et al. Allicin ameliorates imiquimod-induced psoriasis-like skin inflammation via disturbing the interaction of keratinocytes with IL-17A[J]. Br J Pharmacol， 2023， 180(5)： 628-646.
     [5]Griffiths CEM， Armstrong AW， Gudjonsson JE， et al. Psoriasis[J]. Lancet， 2021， 397(10281)： 1301-1315.
     [6]Hawkes JE， Chan TC， Krueger JG. Psoriasis pathogenesis and the development of novel targeted immune therapies[J]. J Eur Acad Dermatol Venereol， 2017， 140(3)： 645-653.
     [7]Wang Z， Zhou H， Zheng H， et al. Autophagy-based unconventional secretion of HMGB1 by keratinocytes plays a pivotal role in psoriatic skin inflammation[J]. Autophagy， 2021， 17(2)： 529-552.
     [8]Zhang C， Xiao C， Dang E， et al. CD100-Plexin-B2 Promotes the Inflammation in Psoriasis by Activating NF-κB and the Inflammasome in Keratinocytes[J]. J Invest Dermatol， 2018， 138(2)： 375-383.
     [9]Wang Y， Zhao N， Yang D， et al. Metabolic reprogramming and AMPK activation： Key players in the therapeutic effects of Cooling Blood and Detoxicating Formular on psoriasis[J]. J Ethnopharmacol， 2025， 337(Pt 1)： 118825.
     [10]El Charif MH， Doughan S， Kredly R， et al. MEBO versus topical Diltiazem versus a combination of both ointments in the treatment of acute anal fissure： a randomized clinical trial protocol[J]. BMC Complement Med Ther， 2021， 21(1)：75.
     [11]庹娅，刘小卫，蒋蕾. 高效液相色谱法同时测定湿润烧伤膏中12种成分含量[J]. 中国药业， 2022， 31(16)： 73-77.
     [12]Qin Z， Tang R， Liang J， et al. Berberine， a natural alkaloid： Advances in its pharmacological effects and mechanisms in the treatment of autoimmune diseases[J]. Int Immunopharmacol， 2024， 137： 112422.
     [13]Chen Y， Song S， Wang Y， et al. Topical application of berberine ameliorates imiquimod-induced psoriasis-like dermatitis in BALB/c mice via suppressing JAK1/STAT1 signaling pathway[J]. Arab J Chem， 2024， 17(3)： 105612.
     [14]Huang KF， Ma KH， Liu PS， et al. Baicalein increases keratin 1 and 10 expression in HaCaT keratinocytes via TRPV4 receptor activation[J]. Exp Dermatol， 2016， 25(8)： 623-629.
     [15]Sun S， Zhang X， Xu M， et al. Berberine downregulates CDC6 and inhibits proliferation via targeting JAK-STAT3 signaling in keratinocytes[J]. Cell Death Dis， 2019， 10(4)： 274.
     [16]Xia C， Fu X， Wang Q， et al. Anti-ROS and NIR-Ⅱ-Responsive Hyaluronic Acid Microneedle Loaded With Baicalin Nanoparticles for Treatment of Psoriasis[J]. Macromol Rapid Commun， 2024， 45(15)： e2400136.
     [17]Gao J， Chen F， Fang H， et al. Daphnetin inhibits proliferation and inflammatory response in human HaCaT keratinocytes and ameliorates imiquimod-induced psoriasis-like skin lesion in mice[J]. Biol Res， 2020， 53(1)： 48.
     [18]Lowes MA， Russell CB， Martin DA， et al. The IL-23/T17 pathogenic axis in psoriasis is amplified by keratinocyte responses[J]. Trends Immunol， 2013， 34(4)： 174-181.
     [19]Hawkes JE， Chan TC， Krueger JG. Psoriasis pathogenesis and the development of novel targeted immune therapies[J]. J Allergy Clin Immunol， 2017， 140(3)： 645-653.
     [20]Fits LVD， Mourits S， Voerman JSA， et al. Imiquimod-induced psoriasis-like skin inflammation in mice is mediated via the IL-23/IL-17 axis[J]. J Immunol， 2009， 182(9)：5836-5845.
     [21]Zhang S， Liu X， Mei L， et al. Epigallocatechin-3-gallate (EGCG) inhibits imiquimod-induced psoriasis-like inflammation of BALB/c mice[J]. BMC Complement Altern Med， 2016， 16(1)： 334.
     [22]Schn MP， Schn M. Imiquimod： mode of action[J]. Br J Dermatol， 2007， 157(Suppl 2)：8-13.
     [23]Saggini A， Chimenti S， Chiricozzi A. IL-6 as a druggable target in psoriasis： focus on pustular variants[J]. J Immunol Res， 2014： 964069.
     [24]Cai Y， Xue F， Quan C， et al. A Critical Role of the IL-1β–IL-1R Signaling Pathway in Skin Inflammation and Psoriasis Pathogenesis[J]. J Invest Dermatol， 2019， 139(1)： 146-156.
     [25]Guo J， Zhang H， Lin W， et al. Signaling pathways and targeted therapies for psoriasis[J]. Signal Transduct Target Ther， 2023， 8(1)： 437.
     [26]Conrad C， Gilliet M. Psoriasis： from Pathogenesis to Targeted Therapies[J]. Clin Rev Allergy Immunol， 2018， 54(1)： 102-113.
     [27]Zhou X， Chen Y， Cui L， et al. Advances in the pathogenesis of psoriasis： from keratinocyte perspective[J]. Cell Death Dis， 2022， 13(1)： 1-13.
     [28]Deng Y， Chang C， Lu Q. The Inflammatory Response in Psoriasis： a Comprehensive Review[J]. Clin Rev Allergy Immunol， 2016， 50(3)： 377-389.
     [29]Zhao W， Xiao S， Li H， et al. MAPK Phosphatase-1 Deficiency Exacerbates the Severity of Imiquimod-Induced Psoriasiform Skin Disease[J]. Front Immunol， 2018， 9： 569.
     [30]Yu XJ， Li CY， Dai HY， et al. Expression and localization of the activated mitogen-activated protein kinase in lesional psoriatic skin[J]. Exp Mol Pathol， 2007， 83(3)：413-418.
     [31]O′Neill LA J， Hardie DG. Metabolism of inflammation limited by AMPK and pseudo-starvation[J]. Nature， 2013， 493(7432)： 346-355.
     [32]Akinduro O， Sully K， Patel A， et al. Constitutive Autophagy and Nucleophagy during Epidermal Differentiation[J]. J Invest Dermatol， 2016， 136(7)： 1460-1470.
     [33]Xiao Y， Jing D， Zhou G， et al. Adenosine 5′monophosphate-activated protein kinase activation reduces the risks of psoriasis and its comorbidities： a Mendelian randomization study in the UK Biobank[J]. Rheumatology， 2024， 63(6)： 1664-1671.
     [34]Jeong HW， Hsu KC， Lee JW， et al. Berberine suppresses proinflammatory responses through AMPK activation in macrophages[J]. Am J Physiol-Endocrinol Metab， 2009， 296(4)： E955-964.