二甲双胍通过NLRP3炎症小体通路对皮肤角质形成细胞增殖和凋亡的双向调节研究  

Bidirectional Regulation of Keratinocyte Proliferation and Apoptosis by Metformin via NLRP3 Inflammasome Pathway

作　　者：田珂[1] 冷秋枫 吕晶 苗国英[1] 王新慧 谢辉 刘渠 姚春霞[3] TIAN Ke;LENG Qiufeng;LYU Jing;MIAO Guoying;WANG Xinhui;XIE Hui;LIU Qu;YAO Chunxia(Department of Dermatology,Affiliated Hospital of Hebei University of Engineering,Handan 056002,China;Department of Physiology,Medical College,Hebei University of Engineering,Handan 056038,China;Department of Pathology,Medical College,Hebei University of Engineering,Handan 056038,China)

机构地区：[1]河北工程大学附属医院皮肤科,河北省邯郸市056002 [2]河北工程大学医学院生理教研室,河北省邯郸市056038 [3]河北工程大学医学院病理教研室,河北省邯郸市056038

出　　处：《中国全科医学》2025年第6期742-750,共9页Chinese General Practice

基　　金：河北省科技厅重点研发计划项目(20377795D);河北省教育厅科技计划项目(ZD2022002);邯郸市科技计划项目(19422083008-67)。

摘　　要：背景扁平苔藓是一种皮肤-黏膜慢性炎症性疾病。因其病因不明,许多患者治疗效果欠佳,严重影响生活质量,有必要深入研究扁平苔藓的发病机制,为其药物筛选提供新靶点。目的探讨二甲双胍通过NLRP3炎症小体通路对皮肤角质形成细胞增殖和凋亡的调控作用。方法实验时间为2020—2023年。体外实验以人永生化角质形成细胞株HaCaT为研究对象,分为4组:对照组、脂多糖(LPS)组(5μg/mL)、二甲双胍组(Met组:10 mmol/L)、LPS与二甲双胍联合组(LPS+Met组:LPS 5μg/mL刺激2 h后,再给予二甲双胍10 mmol/L处理)。体内实验以BALB/c小鼠为研究对象,利用咪喹莫特涂抹小鼠背部皮肤诱导了银屑病样皮炎模型,并制备了二甲双胍乳膏进行治疗,将小鼠随机分为3组:对照组、咪喹莫特组(IMQ组)、咪喹莫特与二甲双胍联合组(IMQ+Met组),每组10只;对照组小鼠于背部涂抹凡士林,IMQ组小鼠于背部涂抹咪喹莫特软膏,IMQ+Met组小鼠于背部涂抹IMQ软膏12 h后再涂抹二甲双胍乳膏;1次/d,连续7 d。采用细胞增殖试剂盒(CCK-8)和流式细胞术检测二甲双胍对HaCaT细胞增殖和凋亡的影响;采用Western Blotting、酶联免疫吸附实验(ELISA)和半胱氨酸天冬氨酸蛋白酶1(Caspase-1)活性实验检测二甲双胍处理HaCaT细胞后NOD样受体蛋白3(NLRP3)炎症小体通路的蛋白表达情况及活性水平;最后采用皮肤组织切片苏木素-伊红(HE)染色和免疫组化检测二甲双胍对咪喹莫特诱导银屑病小鼠皮炎的抗炎效果。结果CCK-8实验结果显示:LPS组、Met组、LPS+Met组HaCaT细胞48 h存活率均低于对照组,而LPS+Met组HaCaT细胞48 h存活率高于LPS组(P<0.05)。流式细胞术结果显示:LPS组、Met组HaCaT细胞48 h G2/M期细胞、细胞凋亡比例均高于对照组,而LPS+Met组HaCaT细胞48 h G2/M期细胞、细胞凋亡比例低于LPS组(P<0.05)。Western Blotting结果显示:LPS组、Met组HaCaT细胞NLRP3炎症小体通路Caspase-1 p40、Background Lichen planus is a chronic inflammatory disease of the skin and mucosa.Due to its unknown etiology,many patients have poor treatment effect,which seriously affects the quality of life.It is necessary to further study the pathogenesis of lichen planus to provide a new target for drug screening.Objective To investigate the effects of metformin on keratinocyte proliferation and apoptosis through NLRP3 inflammasome pathway.Methods Experiment period:2020 to 2023.In vitro experiment,human immortalized keratinocytes(HaCaT)were divided into 4 groups:control group,lipopolysaccharide group(LPS group:5μg/mL),metformin group(Met group:10 mmol/L),LPS combined with metformin group(LPS+Met group:metformin was treated with 10 mmol/L after LPS 5μg/mL stimulation for 2 hours).In vivo experiments,BALB/c mice were used as research objects to induce psoriatic dermatitis model by applying imiquimod on the back skin,and metformin cream was prepared for treatment.The mice were randomly divided into 3 groups:control group,imiquimod group(IMQ group),and imiquimod plus metformin group(IMQ+Met group).Each group has 10 mice.Mice in the control group were smeared with petroleum jelly on the back,mice in the IMQ group were smeared with imiquimod ointment on the back,mice in the IMQ+Met group were smeared with metformin cream after 12 h of IMQ ointment.Once a day for seven consecutive days.Cell counting kit-8(CCK-8)and flow cytometry were used to detect the effects of metformin on the proliferation and apoptosis of HaCaT cells.Western Blotting,enzyme-linked immunosorbent assay(ELISA)and Caspase-1 activity assay were used to detect the expression and activity of NOD-like receptor protein 3(NLRP3)inflammasome pathway protein in HaCaT cells treated with metformin.Finally,hematoxylin-eosin(HE)staining and immunohistochemistry were used to detect the anti-inflammatory effect of metformin on imiquimod-induced psoriatic dermatitis in mice.Results The results of CCK-8 experiment showed that the 48 h survival rate of HaCaT cells in LPS,Met

关 键 词：二甲双胍 NLRP3炎症小体 HACAT细胞 细胞增殖 细胞凋亡 扁平苔藓 银屑病 小鼠 

分 类 号：R977.15[医药卫生—药品] R329.25[医药卫生—药学]