Effect of MIF antisense oligonucleotides on expression of MIF in macrophages |
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Authors: | LI Guang-ran CHEN Wei-ying YU Xue-qing LI Xiao-yan YANG Xiao |
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Affiliation: | Department of Nephrology, The First Affiliated Hospital of Sun Yat-sen University, Key Laboratory of Nephrology in Ministry of Education, Guangzhou 510080, China |
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Abstract: | 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 MIF 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 oligonucleotides of MIF inhibit the expression of MIF mRNA and MIF protein in macrophages treated with LPS. |
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