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...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。展开更多
Macrophage cells play an important role in the initiation and regulation of the immune response. All-trans retinoic acid (ATRA) and its natural and synthetic analogs (retinoids) affect a large number of biological pro...Macrophage cells play an important role in the initiation and regulation of the immune response. All-trans retinoic acid (ATRA) and its natural and synthetic analogs (retinoids) affect a large number of biological processes.Recently , retinoids have been shown promise in the therapy and prevention of various cancers. However, many interesting questions related to the activities of retinoids remain to be answered: (Ⅰ) Molecular mechanisms by which retinoids exert their effects; (Ⅱ) why the clinical uses of retinoids give undesirable side effects of varying severity with a higher freqllency of blood system symptoms; (Ⅲ)little is known for its impacts on macrophage cells etc. We set up this experiment, therefore, to examine the apoptosis of ATRA on macrophage Ana-1 cell line. Apoptosis of the cells was quantitated, after staining cells with propidium iodide (PI), by both accounting nuclear condensation and flow cytometry. When the cells were treated with ATRA at or higher than 1 μM for more than 24 h, significant amount of the apoptotic cells was observed. Induction of apoptosis of Ana-1 cells by ATRA was in time- and dose-dependent manners, exhibiting the similar pattern as the apoptosis induced by actinomycin D (ACTD). ATRA treatment of Ana-1 cells also caused the changes of the mRNA levels of apoptosis-associated gene bcl-2, as detected by Northern blot analysis. The temporal changes of bcl-2 expression by ATRA was also parallel to that by ACTD. In conclusion,ATRA can induce apoptosis in macrophage cells, which may be helpful in understanding of immunological functions retinoids.展开更多
文摘目的: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。
文摘Macrophage cells play an important role in the initiation and regulation of the immune response. All-trans retinoic acid (ATRA) and its natural and synthetic analogs (retinoids) affect a large number of biological processes.Recently , retinoids have been shown promise in the therapy and prevention of various cancers. However, many interesting questions related to the activities of retinoids remain to be answered: (Ⅰ) Molecular mechanisms by which retinoids exert their effects; (Ⅱ) why the clinical uses of retinoids give undesirable side effects of varying severity with a higher freqllency of blood system symptoms; (Ⅲ)little is known for its impacts on macrophage cells etc. We set up this experiment, therefore, to examine the apoptosis of ATRA on macrophage Ana-1 cell line. Apoptosis of the cells was quantitated, after staining cells with propidium iodide (PI), by both accounting nuclear condensation and flow cytometry. When the cells were treated with ATRA at or higher than 1 μM for more than 24 h, significant amount of the apoptotic cells was observed. Induction of apoptosis of Ana-1 cells by ATRA was in time- and dose-dependent manners, exhibiting the similar pattern as the apoptosis induced by actinomycin D (ACTD). ATRA treatment of Ana-1 cells also caused the changes of the mRNA levels of apoptosis-associated gene bcl-2, as detected by Northern blot analysis. The temporal changes of bcl-2 expression by ATRA was also parallel to that by ACTD. In conclusion,ATRA can induce apoptosis in macrophage cells, which may be helpful in understanding of immunological functions retinoids.
