LncRNA IFNG-AS1对巨噬细胞分化及炎症水平的影响

Effects of LncRNA IFNG-AS1 on Macrophage Proliferation and Apoptosis and the Differentiation of Inflammatory Cytokines

目的 长链非编码RNA(LncRNA)IFNG-AS1与冠心病炎症细胞因子水平相关,本研究旨在探讨LncRNA IFNG-AS1对冠心病粥样斑块形成的主要成分巨噬细胞的增殖、凋亡分化及炎症水平的影响。方法 通过PMA将单核细胞诱导为巨噬细胞,并将诱导后的细胞分为不同组进行干预,M0组为仅诱导,M1组为诱导加炎症刺激,M1+Negative Control组为M1的基础上转染阴性对照ASO,M1+ASO-IFNG-AS1组为M1的基础上加沉默该LncRNA的ASO。并检测四组不同处理细胞的增殖、凋亡、迁移及分化情况,并检测炎症细胞因子及NF-κB炎症信号通路激活情况。结果 M1+ASO-IFNG-AS1组巨噬细胞增殖、迁移比例降低,凋亡比例明显升高,且M1+ASO-IFNG-AS1组巨噬细胞倾向于M2型分化。ELISA及WB结果显示,TNF-α、IL-6水平M1+ASO-IFNG-AS1组低于M1组、M1+Negative Control组(P<0.05)。IL-10的水平M1组、M1+Negative Control组低于M0组及M1+ASO-IFNG-AS1组(P<0.05)。p50mRNA表达水平M1+ASO-IFNG-AS1组水平低于M1组、M1+Negative Control组(P<0.05);p-p65以及p50蛋白表达水平,M1组、M1+Negative Control组及M1+ASO-IFNG-AS1组均高于M0组(P<0.05),其中M1+ASO-IFNG-AS1组水平低于M1组、M1+Negative Control组(P<0.05)。在LncRNA IFNG-AS1被抑制后细胞炎症细胞因子IL-6、TNF-α表达降低,抑制炎症的细胞因子IL-10表达增加,p50表达水平降低,p65磷酸化水平降低。结论 LncRNA IFNG-AS1被沉默后可抑制巨噬细胞增殖、迁移,促进凋亡并抑制NF-κB信号通路的激活降低炎症细胞因子表达水平。

Objective To investigate LncRNA IFNG-AS1 effects on the proliferation, apoptosis and differentiation of macrophages as well as inflammation level.Methods Monocytes were induced into macrophages by PMA,and induced cells were divided into different groups for intervention.M0 group was only induced, M1 group was induced with inflammatory stimulation, and M1+ Negative Control group was transfected with negative control ASO on the basis of M1.M1+ ASO-IFnG-AS1 group was ASO of LncRNA silenced on the basis of M1.The proliferation, apoptosis, migration and differentiation of four groups of different treatment cells were detected, and activation of inflammatory cytokines and NF-κB inflammatory signaling pathway was detected.Results Proliferation and migration ratio of macrophages in M1+ASO-IFNG-AS1 group decreased, while apoptosis ratio increased significantly, and macrophages in M1+ASO-IFNG-AS1 group tended to be M2-type differentiation. ELISA and WB results showed that the levels ofTNF-a andIL-6 in the M1+ASO-IFNG-AS1 group were lower than those in the Ml group and M1+Negative Control group(P<0.05). The levels of IL-10 in the M1 group and M1+Negative Control group were lower than those in the MO group and M1+ASO-IFNG-AS1 group(P<0.05).The levels of p50mRNA in the M+ASO-IFNG-AS1 group were lower than those in the Ml group and M1+Negative Control group(P<0.05);p-p65 as well as p50 protein expression levels were higher in theM1group, M1+Negative Control group and M1+ASO-IFNG-AS1 group than in the Mogroup(P<0.05),with lower levels in the M1+ASO-IFNG-AS1 group than in the MI group and M1+Negative Control group(P<0.05). After LncRNA IFNG-AS1 was inhibited, inflammatory cytokines IL-6 and TNF-α decreased, inflammatory cytokines IL-10 increased, p50 decreased and phosphorylation of p65 decreased.Conclusion LncRNA IFNG-AS1 could inhibit proliferation and migration of macrophages, promote apoptosis, inhibit activation of NF-κB signaling pathway and reduce inflammatory cytokines.