MIF反义寡核苷酸对巨噬细胞表达MIF的影响

Effect of MIF antisense oligonucleotides on expression of MIF in macrophages

目的:研究巨噬细胞移动抑制因子(MIF)反义寡核苷酸对体外巨噬细胞表达MIF的影响。方法:设计特异性MIF寡核苷酸,其序列为:反义:5’-TACGGATACAAGTAGCAC-3’;正义:5’-ATGCCTATGTTCATCGTG-3’;错义:5’-CTCTCAGACTCGATCTGT-3’。通过脂质体包裹后将其分别转染至小鼠巨噬细胞,观察MIF反义寡核苷酸对脂多糖(LPS)刺激巨噬细胞表达MIF的影响。结果:LPS刺激后6h,巨噬细胞表达MIFmRNA和合成分泌的MIF量增加,9-12h达高峰。MIF反义寡核苷酸转染巨噬细胞后,可见LPS刺激的MIFmRNA和MIF的表达均明显少于LPS刺激组、LPS刺激+转染正义寡核苷酸组和LPS刺激+转染错义寡核苷酸组(P<0.05),与非LPS刺激组无显著差异。结论:LPS可刺激小鼠巨噬细胞合成和表达MIF。MIF反义寡核苷酸可显著抑制巨噬细胞MIF的过度表达。

AIM： To investigate the effect of antisense oligonucleotides on expression of macrophage migration inhibitory factor （MIF） in macrophages. METHODS： MIF phosphorothioate oligonucleotides was designed and synthesized. The phosphorothioate antisense, sense and missense oligonucleotides of mouse MIF was transfected into macrophages, separately. After that, macrophages were incubated with LPS. Cell culture medium was collected for MIF protein detection by EIA. Cellular RNA was extracted and the expression of MIF mRNA was examined by RT- PCR analysis. RESULTS： LPS stimulation resulted in a specific time- dependent expression of MW derived from macrophages. MIF mRNA and MIF protein level increased at 6 h and reached a plateau at 9 - 12 h after LPS stimulation. The macrophages treated with antisense oligonucleotides showed a significant decrease in MIF mRNA and MIF protein after LPS stimulation than those with LPS stimulation only and LPS plus sense or missense oligonucleotides （ P 〈 0.05）. CONCLUSION： Antisense ohgonucleotides of MIF inhibit the expression of MIF mRNA and MIF protein in macrophages treated with LPS.