牙髓成纤维细胞内质网应激在牙髓炎中的作用

doi:10.13701/j.cnki.kqyxyj.2022.11.013

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (11): 1064-1070.DOI: 10.13701/j.cnki.kqyxyj.2022.11.013

牙髓成纤维细胞内质网应激在牙髓炎中的作用

李丹^1,2, 张旗^1,2*

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

收稿日期:2022-04-26 出版日期:2022-11-25 发布日期:2022-11-22
通讯作者: *张旗,E-mail:qizhang@tongji.edu.cn
作者简介:李丹(1994~ ),女,甘肃人,硕士在读,研究方向:牙髓炎活髓保存治疗。
基金资助:
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)

Effects of Dental Pulp Fibroblasts Endoplasmic Reticulum Stress in Pulpitis

LI Dan^1,2, ZHANG Qi^1,2*

1. Department of Endodontics, School and Hospital of Stomatology, Tongji University, Shanghai 200072, China;
2. Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China

Received:2022-04-26 Online:2022-11-25 Published:2022-11-22

摘要/Abstract

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。

关键词: 牙髓炎, 牙髓成纤维细胞, 内质网应激

Abstract: 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.

Key words: pulpitis, dental pulp fibroblasts, endoplasmic reticulum stress

李丹, 张旗. 牙髓成纤维细胞内质网应激在牙髓炎中的作用[J]. 口腔医学研究, 2022, 38(11): 1064-1070.

LI Dan, ZHANG Qi. Effects of Dental Pulp Fibroblasts Endoplasmic Reticulum Stress in Pulpitis[J]. Journal of Oral Science Research, 2022, 38(11): 1064-1070.

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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

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