牙髓成纤维细胞内质网应激在牙髓炎中的作用  

Effects of Dental Pulp Fibroblasts Endoplasmic Reticulum Stress in Pulpitis

作　　者：李丹 张旗[1,2] LI Dan;ZHANG Qi(Department of Endodontics,School and Hospital of Stomatology,Tongji University,Shanghai 200072,China;Shanghai Engineering Research Center of Tooth Restoration and Regeneration,Shanghai 200072,China)

机构地区：[1]同济大学口腔医学院·同济大学口腔医院牙体牙髓科,上海200072 [2]上海牙组织修复与再生工程技术研究中心,上海200072

出　　处：《口腔医学研究》2022年第11期1064-1070,共7页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81870760、82170945);临床“五新”创新研发项目(编号:SHDC2020CR3058B)。

摘　　要：目的:探究内质网应激(endoplasmic reticulum stress,ERS)在牙髓炎症进程中的发生和作用,以及对人牙髓成纤维细胞(human dental pulp fibroblasts,hDPFs)炎症因子的影响。方法:构建大鼠牙髓炎模型,0、6、12、24、48 h取材。苏木素-伊红染色观察炎症浸润;RT-qPCR检测ERS及炎症因子相关mRNA表达水平。体外培养hDPFs,分对照组、4-苯基丁酸(4-phenylbutyric acid,4-PBA)组、LPS组、LPS+4-PBA组,1 d收样,透射电镜观察内质网形态;免疫荧光检测ERS标志蛋白表达;RT-qPCR检测ERS标志物及炎症因子mRNA表达。体内髓腔应用4-PBA进行“挽救”,1、3、7 d收样。结果:随暴露时间延长,牙髓组织炎症浸润增加,TNF-α、IL-1β、IL-6和GRP78、CHOP、XBP1 mRNA表达升高(P<0.05)。体外,LPS组内质网形态发生肿胀、扩张,GRP78、CHOP蛋白表达增加,TNF-α、IL-1β、IL-6和GRP78、CHOP、XBP1 mRNA表达升高,4-PBA改善内质网损伤,GRP78、CHOP表达降低,促炎因子mRNA表达降低(P<0.05)。体内“挽救”表明4-PBA减轻炎症浸润,抑制TNF-α、IL-1β、GRP78、CHOP表达(P<0.05)。结论:牙髓炎症进程中,伴随ERS发生和炎症因子分泌增加,4-PBA抑制ERS,减轻炎症反应。Objective:To explore the occurrence and effects of endoplasmic reticulum stress(ERS)in pulpitis and on inflammatory factor of human dental pulp fibroblasts(hDPFs).Methods:Rats were used to construct the pulpitis models.The samples were collected at 0,6,12,24,and 48 hours.Hematoxylin-eosin staining was used to observe the inflammatory infiltration.The mRNA expression levels of ERS and inflammatory factors were detected by RT-qPCR.HDPFs were cultured in vitro and divided into control group,4-phenylbutyric acid(4-PBA)group,LPS group,and LPS+4-PBA group.The samples were collected at 1 d.Endoplasmic reticulum morphology was observed by transmission electron microscopy.The expression of ERS markers proteins were detected by immunofluorescence.The mRNA expression of ERS makers and inflammatory factors were detected by RT-qPCR.4-PBA was used in the pulp cavity for"rescue",and samples were collected at 1,3,and 7 d.Results:With increased exposure time,the inflammatory infiltration of pulp tissue increased,and the mRNA expressions of TNF-α,IL-1β,IL-6,GRP78,CHOP,and XBP1 increased(P<0.05).In vitro,the endoplasmic reticulum was swelling and dilation in LPS group,and the expressions of GRP78 and CHOP proteins were increased.The mRNA expressions of TNF-α,IL-1β,IL-6,GRP78,CHOP and XBP1 were increased.4-PBA improved endoplasmic reticulum injury,decreased GRP78 and CHOP expression,and decreased mRNA expression of pro-inflammatory factors(P<0.05).In vivo"rescue"showed that 4-PBA could reduce the inflammatory infiltration and inhibit the expression of TNF-α,IL-1β,GRP78,and CHOP(P<0.05).Conclusion:In the occurrence and development of pulpitis,ERS is involved and increased the secretion of inflammatory factors.4-PBA inhibits ERS and improves inflammatory response.

关 键 词：牙髓炎 牙髓成纤维细胞 内质网应激 

分 类 号：R781.31[医药卫生—口腔医学]