摘要
目的:探究姜黄素(Cur)对牙龈卟啉单胞菌(Pg)脂多糖(LPS)诱导的人牙龈成纤维细胞(HGFs)炎症反应及微小RNA-124(miR-124)的影响。方法:将HGFs分为对照(control)组、LPS组(10 mg/L LPS)及LPS+Cur(20、40和80μmol/L)组(10 mg/L LPS+对应剂量Cur)。各组处理24 h,CCK-8法检测细胞活力;ELISA检测上清液中炎症因子白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的水平;RT-qPCR法检测细胞中miR-124的水平;Western blot检测胞浆和胞核中核因子κB(NF-κB)p-p65蛋白水平;激光共聚焦显微镜检测NF-κB p-p65蛋白的入核情况。转染mimic-NC和miR-124 mimic,检测细胞中miR-124及胞浆和胞核中NF-κB p-p65蛋白水平。结果:LPS组细胞活力、miR-124水平和胞浆NF-κB p-p65蛋白水平低于control组(P<0.05),上清液IL-1β和TNF-α水平及胞核NF-κB p-p65蛋白水平高于control组(P<0.05)。LPS+Cur(40和80μmol/L)组细胞活力、miR-124水平及胞浆NF-κB p-p65蛋白水平高于LPS组(P<0.05),上清液TNF-α水平和胞核NF-κB p-p65蛋白水平低于LPS组(P<0.05)。LPS+80μmol/L Cur组细胞上清液IL-1β水平低于LPS组(P<0.05)。miR-124 mimic组细胞miR-124和胞浆中NF-κB p-p65蛋白水平高于LPS组和mimic-NC组(P<0.05),胞核NF-κB p-p65蛋白水平低于LPS组和mimic-NC组(P<0.05)。结论:Cur可抑制Pg LPS诱导的HGFs炎症反应,可能是通过上调miR-124进而抑制NF-κB p-p65入核实现的。
AIM:To investigate the effects of curcumin(Cur)on the inflammatory response of human gingival fibroblasts(HGFs)induced by Porphyromonas gingivalis(Pg)lipopolysaccharide(LPS)and the role of microRNA-124(miR-124)in this process.METHODS:The HGFs were divided into control group,LPS group(10 mg/L LPS)and LPS+Cur(20,40 and 80μmol/L)groups(10 mg/L LPS+corresponding dose of Cur).After treatment for 24 h,CCK-8 assay was used to measure the cell viability.ELISA was used to measure the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the supernatant.The level of miR-124 in the cells was detected by RT-qPCR.The protein levels of nuclear factor kappa B(NF-κB)p-p65 in cytoplasm and nucleus were determined by Western blot,and the nuclear trans⁃location of NF-κB p-p65 was evaluated by laser confocal microscopy.After transfection with mimic-NC or miR-124 mimic,the expression of miR-124 and NF-κB p-p65 protein in the cytoplasm and nucleus of the cells were also detected.RESULTS:The cell viability,the level of miR-124 in the cells and NF-κB p-p65 protein level in cytoplasm of LPS group were lower than those in control group(P<0. 05),while the levels of IL-1β and TNF-α in the supernatant and NF-κB pp65protein level in the nucleus were higher than those in control group(P<0. 05). The cell viability,the level of miR-124 in cells and NF- κB p-p65 protein level in the cytoplasm of LPS+Cur(40 and 80 μmol/L)groups were higher thanthose in LPS group(P<0. 05),while the level of TNF-α in the supernatant and NF-κB p-p65 protein level in the nucleuswere lower than those in LPS group(P<0. 05). The level of IL-1β in the supernatant of LPS+80 μmol/L Cur group waslower than that in LPS group(P<0. 05). The levels of miR-124 and NF-κB p-p65 protein level in the cytoplasm of miR-124 mimic group were higher than those in LPS group and mimic-NC group(P<0. 05),while the level of NF-κB p-p65 pro⁃teinlevel in the nucleus was lower than that in LPS group and mimic-NC group(P<0. 05). CONCLUSION:Curcumin in⁃hibits the inflammatory response of HGFs induced by Pg LPS,which may be achieved by up-regulating miR-124 and theninhibiting the nuclear translocation of NF-κB p-p65.
作者
李坤阳
陈栋
左春然
刘爱群
朱兰省
牛兵
LI Kun-yang;CHEN Dong;ZUO Chun-ran;LIU Ai-qun;ZHU Lan-sheng;NIU Bing(Department of Stomatology,Henan Province Hospital of Traditional Chinese Medicine(The Second Affiliated Hospital of Henan University of Traditional Chinese Medicine),Zhengzhou 450001,China;Department of Stomatology,The First Af-filiated Hospital of Zhengzhou University(Henan Province Stomatologic Hospital),Zhengzhou 450052,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第10期1867-1874,共8页
Chinese Journal of Pathophysiology
基金
2018年度河南省医学科技攻关计划项目(No.SBGJ2018038)
河南省高等学校重点科研项目计划(No.18A310036)。
