HSV Us3蛋白激酶对巨噬细胞生物学活性和凋亡的影响研究

Effects of HSV Us3 Protein Kinase on Bio-Activity and Apoptosis of Megaphage

为探讨单纯疱疹病毒Us3基因及其编码产物对LPS活化的巨噬细胞生物学活性和凋亡的影响,小鼠巨噬细胞系RAW246.7细胞瞬时转染pcDNA3.1(+)-Us3重组质粒并体外培养。利用免疫荧光技术检测转染率,ELISA方法检测培养上清中TNF-α分泌水平,Annexin-V染色评价转染细胞凋亡情况。结果发现,pcDNA3.1(+)-Us3重组质粒可成功转染至RAW246.7细胞,转染率为10.3%。与空质粒转染组相比,pcDNA3.1(+)-Us3转染细胞培养上清中TNF-α分泌水平明显升高,但凋亡细胞数量略有下降,证实Us3基因及其编码蛋白在调控巨噬细胞细胞因子分泌活性及其活化细胞凋亡方面发挥一定作用。

In order to investigate the effect of Us3 gene of protein kinase of herpes simplex virus（ HSV） on regulating cytokine production and apoptosis of RAW246. 7 cells after LPS activation,RAW246. 7 cells were transfected with recombinant plasmid of pcDNA3. 1（ ＋）-Us3 and cultured in vitro. Immunofluorescence staining was used for identification of transfection rate. The level of TNF-α in the cultured supernatant was detected by ELISA. And evaluated apoptosis of transfected cells with Annexin-V staining. The results showed that recombinant plasmid pcDNA3. 1（ ＋）-Us3 could successfully transfected into RAW246. 7 with transfection rate at 10. 3%. As compared with empty plasmid transfected group,the secretion level of TNF-α in supernatant transfected cells by pcDNA3. 1（ ＋）-Us3 was apparently ascent,but the amount of apoptosis cells were slightly descent,thus proved that Us3 gene and it coded protein play a certain role in regulating and controlling cytokine secretive activity by macrophage as well as in activating cell apoptosis.