重组HMGB1对DEN2感染Ana-1细胞分泌TNF-α及NO的影响

Effect of HMGB1 on secretion of inflammatory cellular factor TNF-α and NO in mice macrophage infected with DEN2

目的:观察不同浓度重组高迁移率族蛋白1(recombination high mobility group box protein 1,r HMGB1)对DEN2感染鼠源单核巨噬细胞Ana-1分泌TNF-α、NO及DEN复制的影响。方法:直接免疫荧光法鉴定DEN2吸附Ana-1细胞;qRTPCR检测不同时间胞内病毒载量变化;双抗体夹心ELISA法检测感染不同时间TNF-α的分泌水平,Griess试剂检测细胞释放NO变化。结果:不同剂量r HMGB1处理Ana-1细胞后,对DEN2感染细胞的病毒抑制率(%)分别为41.53±2.12﹑55.30±1.59﹑74.75±1.12﹑86.35±1.42,差异有统计学意义(P<0.05)。r HMGB1处理后,感染细胞分泌TNF-α和NO水平均有所下降,差异有统计学意义(P<0.05)。结论:r HMGB1可有效调节DEN2感染Ana-1细胞的TNF-α和NO分泌,并抑制DEN2的增殖。

Objective： To observe the effect of different concentration HMGB1 on the secretion of TNF-α and NO from Ana-1infected with DEN2 and virus copy. Methods： DEN2 were proliferated and identified by conventional methods. The adherence of DEN2 to Ana-1 was observed by direct immunofluorescence and RT-PCR. The level of virus mRNA were detected by qRT-PCR. The concentration of TNF-α was detected by ELISA. The concentration of NO was detected with Griess reagent. Results： Ana-1 was able to adhered for DEN2. Compared with DEN group,the inhibition ratio（ %） of the level of virus mRNA in D-HMGB1-1 group,D-HMGB1-10 group,D-HMGB1-100 group,D-HMGB1-1000 group was 41. 53 ± 2. 12,55. 30 ± 1. 59,74. 75 ± 1. 12,86. 35 ± 1. 42. Compared with DEN group,the level of TNF-α and NO decreased in D-HMGB1 groups（ P 〈0. 05）. Conclusion： HMGB1 can be effectively regulated of Ana-1 secreted inflammation factor of infected with DEN2,and inhibited DEN2 replication.