目的:以脂多糖(LPS)处理的人牙髓细胞(HDPCs)为细胞模型,研究微小RNA(miR)-152在牙髓炎发病中的作用。
方法:使用细胞计数试剂盒(CCK-8)测量LPS感染的HDPC的生物活性,Transwell测试,流式细胞术,和荧光定量PCR。使用测定试剂盒评估超氧化物歧化酶(SOD)和丙二醛(MDA)的浓度,酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-1β和IL-6水平,并通过双荧光素酶报告检测SMAD5与miR-152的靶向关系。通过PCR评估细胞周期相关的CyclinD1和BAX的表达。通过绘制受试者工作特性(ROC)曲线,显示了miR-152的诊断价值.
结果:LPS诱导的HDPCs中miR-152水平降低,而SMAD5水平升高。在LPS诱导的HDPC中过表达miR-152后,生存能力提高了,细胞凋亡率下降,CyclinD1升高,BAX减少了,炎症细胞因子(IL-6和IL-1β)被抑制,SOD活性增加,MDA含量降低。miR-152靶向调控SMAD5,SMAD5调控miR-152对细胞活力的影响,凋亡,炎症,和HDPC的氧化反应。在牙髓炎患者中证实miR-152表达降低,这可能是牙髓炎的生物标志物。
结论:miR-152被发现是与牙髓炎发病机制和HDPC生物学行为相关的生物标志物。
OBJECTIVE: To research the role of microRNA (miR)-152 in the pathogenesis of pulpitis using a cell model based on human dental pulp cells (HDPCs) treated with lipopolysaccharides (LPS).
METHODS: The biological activity of HDPCs infected by LPS was measured using a cell counting kit (CCK-8), Transwell test, flow cytometry, and fluorescent quantitative PCR. The concentration of superoxide dismutase (SOD) and malondialdehyde (MDA) was evaluated using an assay kit, the levels of interleukin (IL)-1β and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA), and the targeting relationship between SMAD5 and miR-152 was measured by the double-luciferase report test. The expression of cell cycle-related CyclinD1 and BAX was assessed by PCR. By plotting a receiver operating characteristic (ROC) curve, the diagnostic value of miR-152 was shown.
RESULTS: The level of miR-152 in HDPCs induced by LPS decreased, while the level of SMAD5 increased. After overexpressing miR-152 in LPS-induced HDPCs, the viability was elevated, the apoptosis rate decreased, CyclinD1 was elevated, BAX diminished, the inflammatory cytokines (IL-6 and IL-1β) were inhibited, the activity of SOD increased, and the MDA content decreased. miR-152 targeted regulation of SMAD5, and SMAD5 modulated the effects of miR-152 on cell viability, apoptosis, inflammation, and the oxidative response of HDPCs. Reduced miR-152 expression was verified in patients with pulpitis, which could be a biomarker for pulpitis.
CONCLUSIONS: miR-152 was found to be a biomarker correlated with the pathogenesis of pulpitis and the biological behaviour of HDPCs.