长链非编码RNA-EPS对小鼠牙髓细胞凋亡的影响

Effects of long non-coding RNA-EPS on apoptosis of mouse dental pulp cells in vitro

目的:研究长链非编码RNA-EPS(long noncoding RNA-EPS,lnc-EPS)对炎症状态下小鼠牙髓细胞凋亡的影响。方法:分离和培养小鼠牙髓细胞(mouse dental pulp cells,mDPCs),利用实时荧光定量PCR(qRT-PCR)检测脂多糖(lipopolysaccharide,LPS)刺激情况下,lnc-EPS的表达情况。应用lnc-EPS过表达质粒和小干扰RNA(siRNA)影响lnc-EPS的表达后,采用qRT-PCR检测不同组细胞中NLRP3炎症小体(NLRP3、ASC、Caspase-1)相关基因和白介素-1β(IL-1β)的表达情况。采用Western Blot检测细胞NLRP3炎症小体蛋白的表达情况;采用免疫荧光流式细胞术检测不同组细胞的凋亡情况。结果:在LPS刺激情况下,lnc-EPS的表达明显升高(P<0.05);lnc-EPS沉默后,NLRP3炎症小体和IL-1β的mRNA水平显著升高(P<0.01);NLRP3炎症小体表达量和凋亡率均显著升高(P<0.01)。相反,lnc-EPS过表达后,NLRP3炎症小体蛋白和IL-1β的mRNA水平显著降低(P<0.01);NLRP3炎症小体蛋白表达量和凋亡率均显著降低(P<0.01)。结论:lnc-EPS可抑制小鼠牙髓细胞凋亡。

Objective:To study the effects of long noncoding RNA-EPS(lnc-EPS)on apoptosis of dental pulp cells under inflammatory conditions.Methods:Mouse dental pulp cells(mDPCs)were isolated and cultured,and then the expression of lnc-EPS in the stimulation of lipopolysstimulate(LPS)was detected by quantitative real-time PCR(qRT-PCR).By using lnc-EPS overexpressed plasmid and small interfering RNA(siRNA)to affect the expression of lnc-EPS,the expressions of NLRP3 inflammasome(NLRP3,ASC,caspase-1)related genes and interleukin-1β(IL-1β)in different groups of cells were detected by qRT-PCR.Western Blot was used to detect the expression of NLRP3 inflammasome protein.The apoptosis of cells in different groups was detected by immunofluorescence flow cytometry.Results:The expression of lnc-EPS was significantly increased under LPS stimulation(P<0.05).After the silence of lnc-EPS,the mRNA levels of NLRP3 inflammasome and IL-1β were significantly increased(P<0.01).NLRP3 inflammasome protein expression and apoptosis rate were significantly increased(P<0.01).Conversely,after overexpression of lnc-EPS,the mRNA levels of NLRP3 inflammasome protein and IL-1β were significantly reduced(P<0.01).NLRP3 inflammasome protein expression and apoptosis rate were significantly reduced(P<0.01).Conclusion:Lnc-EPS can suppress the apoptosis of mouse dental pulp cells.