p47 GTPase缺失对小鼠巨噬细胞成熟及功能的影响

Effects of p47 GTPase deficiency on the maturation and function of macrophages in mice

目的以日本血吸虫感染小鼠为模型,研究p47GTP酶家族成员IGTP及IRG-47缺失对巨噬细胞发育成熟及功能的影响。方法常规方法建立日本血吸虫感染小鼠模型,通过实时荧光定量PCR观察日本血吸虫不同感染时期igtp或irg47单一基因缺失的巨噬细胞igtp、lrg47和irg47的mRNA水平;流式细胞术检测巨噬细胞表面分子MHC-Ⅱ类(I-A)、CD16/32、CD40和CD80的表达;应用硝酸还原酶法检测感染急性期巨噬细胞产生炎症因子NO的能力。结果巨噬细胞igtp、irg47及lrg47基因在感染血吸虫后3周表达快速上调,感染后6周上调表达的幅度相差各异,IRG-47-/-小鼠igtp和lrg47转录明显上调。感染前和感染后3周IGTP-/-组的巨噬细胞表面CD16/32和CD40表达均较IRG-47-/-组和WT组高;感染后6周表达差异均无统计学意义(P>0.05)。在感染后6周,无有丝分裂原刺激情况下IGTP-/-组的巨噬细胞较IRG-47-/-组和WT组产生更高水平的NO;LPS刺激后,3组巨噬细胞产生的NO水平差异无统计学意义(P>0.05)。结论IGTP及IRG-47缺失不影响巨噬细胞发育成熟,也不影响巨噬细胞产生炎症因子NO的潜能,提示p47GTP酶可能具有独特的、不依赖NO的作用方式。

Objective To investigate the effects of p47 OTPase deficiency on the maturation and function of macrophages in the IGTP and IRG-47 deficient mice model infected with Schistosoma japonicum. Methods Measured the mRNA levels of igtp, irg47 and lrg47 with IGTP and IRG 47 deficient maerophages by real time quantitative PCR, the expression of surface molecule markers on IGTP and IRG-47 deficient macrophages in different infection stages by FACS, and the inflammatory response By the measurement of NO in the culture supernatant of macrophages stimulated with or without LPS. Results Maerophage mRNA expression levels of igtp, irg47 and lrg4？ were quickly up regulated after 3 weeks of infection, while after 6 weeks of infection they displayed differentiation such that the expression of igtp and lrg47 was significantly up-regulated in IRG-47 / mice. There was no significance in the expression of MHC class II （I-A）, CD16/ 32, CD40 and CD80 on the surface of IGTP and IRG-47 deficient maerophages at 6 weeks after infection. Groups produced equivalent levels of NO after stimulation by LPS. Conclusion IGTP or IRG-47 deficiency does not influence macrophage maturation and the potential for secreting NO, suggesting that p47 GTPase may function in a unique NO-independent manner.