槲皮素通过JAK/STAT信号通路预防吸烟牙周炎患者牙周病变的初步研究  

A preliminary study of quercetin in the prevention of periodontal lesions in smokers through JAK/STAT signal pathway

作　　者：沈益名 齐霞 冯钰皓 杨冬茹[1] SHEN Yiming;QI Xia;FENG Yuhao;YANG Dongru(School and Hospital Stomatology Hebei Medical University,Hospital of Stomatology Hebei Medical University·Preventive Dentistry,Hebei Key Laboratory of Stomatology,The Oral Disease Clinical Medicine Research Institution of Hebei,Shijiazhuang 050017)

机构地区：[1]河北医科大学口腔医学院·口腔医院牙周一科,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,石家庄050017

出　　处：《现代口腔医学杂志》2024年第2期100-107,共8页Journal of Modern Stomatology

基　　金：河北省自然科学基金和重点基础研究专项(H2019206541);河北省省级科技计划(临床医学研究中心专项)(20577717D);科技计划(卫生健康创新专项)(21377715D)。

摘　　要：目的通过建立牙周炎及伴吸烟牙周炎体外模型,探究在炎症环境下槲皮素对人牙周膜成纤维细胞(human periodontal ligament fibroblasts,HPLFs)的迁移能力、Ⅰ型胶原(collagenⅠ,COLⅠ)和Ⅲ型胶原(collagenⅢ,COLⅢ)分泌能力以及炎症因子白介素(interleukin,IL)-1β、肿瘤坏死因子(tumor necrosis factor,TNF)-α、环氧化物酶(cyclooxygenase,COX)-2 mRNA表达水平的影响。并探究槲皮素对JAK/STAT信号通路的影响。方法原代培养并鉴定HPLFs,使用香烟烟雾提取物(cigarette smoke extract,CSE)及脂多糖(lipopolysaccharide,LPS)建立炎症及炎症伴吸烟模型,将细胞分为8组:正常组(N)、CSE组(S)、脂多糖组(L)、脂多糖+JAK/STAT阻断剂组(L+AG490)、脂多糖+CSE组(L+S)、脂多糖+CSE+JAK/STAT阻断剂组(L+S+AG490)、脂多糖+槲皮素组(L+Q)、脂多糖+CSE+槲皮素组(L+S+Q)。通过划痕实验检测细胞迁移能力,ELISA检测细胞胶原分泌能力,RT-PCR检测炎症因子表达水平,免疫荧光染色检测细胞p-STAT3的表达情况。结果与N组相比,L、S、L+S组细胞迁移能力和胶原分泌能力降低(P<0.05),炎症因子表达水平和p-STAT3表达量上升(P<0.05);与L组相比,L+AG490组和L+Q组的细胞迁移能力和胶原分泌能力提高(P<0.05),炎症因子表达水平和p-STAT3表达量降低(P<0.05);与L+S组相比,L+S+AG490组和L+S+Q组的细胞迁移能力和胶原分泌能力提高(P<0.05),炎症因子表达水平和p-STAT3表达量降低(P<0.05)。结论香烟烟雾提取物和脂多糖可抑制HPLFs的迁移能力,诱导炎症因子表达,抑制胶原分泌,促进p-STAT3的表达,而AG490和槲皮素对HPLFs起到保护作用,抑制p-STAT3的表达,依据本实验结果,认为槲皮素可能通过抑制JAK/STAT通路对HPLFs起到保护作用。Objective To explore the effect of quercetin on the ability of cell migration,secretion of collagen I(COL I)and collagen II(COL II),mRNA expression of interleukin(IL)-1β,tumor necrosis factor(TNF)-α,cyclooxygenase(COX)-2 of human periodontal ligament fibroblasts(HPLFs)in an inflammatory environment by establishing inflammation and inflammation with smoking model in vitro.And to explore the effect of quercetin on JAK/STAT signal pathway.Methods HPLFs was cultured and identified,and cigarette smoke extract(CSE)and lipopolysaccharide(LPS)were used to establish inflammation and inflammation with smoking model.The cells were divided into 8 groups:normal group(N),CSE group(S),lipopolysaccharide group(L),lipopolysaccharide+JAK/STAT blocker group(L+AG490),lipopolysaccharide+CSE group(L+S),lipopolysaccharide+CSE+JAK/STAT blocker group(L+S+AG490),lipopolysaccharide+quercetin group(L+Q),lipopolysaccharide+CSE+quercetin group(L+S+Q).The ability of cell migration,collagen secretion,expression of inflammatory cytokines and p-STAT3 were detected by scratch test,ELISA,RT-PCR and immunofluorescence staining respectively.Results Compared with group N,the ability of cell migration and collagen secretion decreased,the expression of inflammatory cytokines and p-STAT3 increased in group L,S and L+S(P<0.05).The ability of cell migration and collagen secretion increased in group L+AG490 and L+Q compared with group L,while the expressionof inflammatory factors and p-STAT3 decreased(P<0.05).The ability of cell migration and collagen secretion increased in group L+S+AG490and L+S+Q compared1with group L+S,while the expression of inflammatory factors and P-STAT3 decreased(P<0.05).Conclusion Cigarette smoke extract and lipopolysaccharide can inhibit the migration ability of HPLFs,induce the expression of inflammatory factors,inhibit collagen secretion and promote the expression of p-STAT3,while AG490 and quercetin protect HPLFs and inhibit the expression of p-STAT3.It is speculated that quercetin may protect HPLFs by inhibiting JAK/STAT pat

关 键 词：人牙周膜成纤维细胞 JAK/STAT信号通路 香烟烟雾提取物 槲皮素 牙周炎 

分 类 号：R781.4[医药卫生—口腔医学]