硝基油酸对大鼠下颌下腺上皮细胞放射损伤的保护机制  

Protective mechanism of nitrooleic acid on submandibular gland cell radiation injury in rats

作　　者：林培琦 骆勤亮 张立刚 黄桂林 唐建宏 张霓霓 Lin Peiqi;Luo Qinliang;Zhang Ligang;Huang Guilin;Tang Jianhong;Zhang Nini(Department of Oral and Maxillofacial Surgery,Affiliated Stomatological Hospital of Zunyi Medical University,Zunyi 563000,Guizhou Province,China;Department of Stomatology,Longyan First Hospital,Longyan 364000,Fujian Province,China;Department of Stomatology,Fifth Affiliated Zhuhai Hospital of Zunyi Medical University,Zhuhai 519110,Guangdong Province,China)

机构地区：[1]遵义医科大学附属口腔医院颌面外科,贵州省遵义市563000 [2]龙岩市第一医院口腔科,福建省龙岩市364000 [3]遵义医科大学第五附属珠海医院口腔科,广东省珠海市519110

出　　处：《中国组织工程研究》2025年第26期5520-5527,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81960204),项目负责人:黄桂林;贵州省临床重点建设项目(黔卫计办涵【2017】24号),项目负责人:黄桂林;贵州省科技计划项目(黔科合基础-ZK[2024]一般339),项目负责人:张霓霓;遵义医科大学“未来临床名医”项目(20211017),项目负责人:张霓霓;贵州省教育厅青年科技人才成长项目(黔教合KY字[2022]291号),项目负责人:张立刚;贵州省卫生健康委科学技术基金项目(gzwjkj2020-1-165),项目负责人:张立刚。

摘　　要：背景:研究发现Nrf2/ARE信号通路潜在激活剂硝基油酸具有低毒可控的特征,对放射性组织损伤具有保护作用。目的:探讨硝基油酸能否通过调节Nrf2/ARE信号通路对下颌下腺上皮细胞放射损伤发挥保护作用。方法:体外培养SD大鼠下颌下腺上皮细胞,CCK-8筛选硝基油酸最佳给药浓度与时间,然后将下颌下腺上皮细胞分为未放射组、放射对照组、硝基油酸组、硝基油酸+ML385(Nrf2/ARE信号通路特异性抑制剂)组以及ML385组,按实验分组用硝基油酸和ML385对下颌下腺上皮细胞进行相应预处理24 h,再给予5 Gy射线放射造模。放射后48 h,CCK-8检测细胞增殖率,实时定量PCR检测细胞内Nrf2、HO-1、NQO1 mRNA表达,实时定量PCR及酶联免疫吸附法检测细胞分泌功能及炎症因子表达,DCFH-DA荧光探针试剂盒检测细胞内活性氧水平。结果与结论:①与放射对照组比较,硝基油酸组大鼠下颌下腺上皮细胞增殖率与分泌功能相关因子水通道蛋白5、α-淀粉酶表达水平升高(P<0.05),Nrf2、HO-1、NQO1 mRNA表达升高(P<0.05),炎症因子白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平下降(P<0.05),活性氧生成减少(P<0.01);②与硝基油酸组比较,硝基油酸+ML385组的细胞增殖率与分泌功能相关因子水通道蛋白5、α-淀粉酶表达水平下降(P<0.05),Nrf2、HO-1、NQO1 mRNA表达下降(P<0.05),炎症因子白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平升高(P<0.05),活性氧生成增多(P<0.05)。结果表明,硝基油酸能够减轻放射所致的大鼠下颌下腺上皮细胞损伤,提高细胞增殖能力和分泌功能,降低细胞内炎性因子表达和活性氧水平,其发挥保护作用可能与激活Nrf2/ARE信号通路存在关联。BACKGROUND:Recent studies have found that Nrf2/ARE signaling pathway activators have the characteristics of low toxicity and control,and have a protective effect against radiation tissue damage.OBJECTIVE:To investigate whether nitrooleic acid can protect submandibular gland epithelial cells from radiation injury by regulating the Nrf2/ARE signaling pathway.METHODS:Rat submandibular gland epithelial cells were cultured in vitro and CCK-8 assay was used to screen the optimal concentration and time of nitrooleic acid administration.Submandibular gland epithelial cells were divided into non-radiation group,radiation control group,nitrooleic acid group,nitrooleic acid+ML385(Nrf2/ARE signaling pathway specific inhibitor)group,and ML385 group.Submandibular gland cells were pretreated with nitrooleic acid and ML385 for 24 hous according to the experimental groups,and then irradiated with 5 Gy radiation to establish the models.At 48 hours after irradiation,CCK-8 assay was used to detect the cell proliferation rate.Real-time quantitative PCR was used to detect the expression of Nrf2,HO-1,and NQO1 mRNA in the cells.Real-time quantitative PCR and enzyme-linked immunosorbent assay were used to detect the cell secretion function and the expression of inflammatory factors.DCFH-DA fluorescent probe kit was used to detect the level of intracellular reactive oxygen species.RESULTS AND CONCLUSION:(1)Compared with the radiation control group,the proliferation rate of submandibular gland epithelial cells and the expression levels of secretion function related factors aquaporin 5 andα-amylase in the nitrooleic acid group of rats increased(P<0.05),and the expression levels of Nrf2,HO-1,and NQO1 mRNA increased(P<0.05),while the expression levels of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-αdecreased(P<0.05),and reactive oxygen species generation reduced(P<0.01).(2)Compared with the nitrooleic acid group,the addition of nitrooleic acid and ML385 group resulted in a decrease in cell proliferation rate

关 键 词：下颌下腺上皮细胞 放射性损伤 硝基油酸 Nrf2/ARE信号通路 电离辐射 辐射损伤 抗氧化应激 工程化细胞 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R782