【摘要】 目的:探讨人白细胞介素-10(human interlenkin-10,hIL-10)质粒转染后的表达产物对RANKL诱导的破骨细胞形成的影响。方法:将hIL-10及相应的空载体质粒在脂质体介导下转染293T细胞,获取含hIL-10蛋白的培养液;通过TRAP染色及骨吸收实验观察转染产物对RANKL诱导的小鼠单核/巨噬细胞RAW264.7分化形成破骨细胞的影响。结果:hIL-10质粒脂质体法瞬时转染293T及RAW264.7细胞获得成功,部分细胞发出绿色荧光,ELISA检测表明目的基因获得表达,RAW264.7细胞在RANKL诱导下可形成破骨细胞样细胞,在该培养体系中加入含hIL-10蛋白的培养液上清,TRAP染色阳性细胞及骨吸收陷窝面积显著减少(P<0.05)。结论:本实验所采用的hIL-10质粒可以成功转染293T及RAW264.7细胞,产生的hIL-10蛋白在体外具有抑制破骨细胞形成和骨吸收的生物学功能。更多还原

【Abstract】 Objective: To explore the effects of gene transfection with plasmid DNA encoding hIL-10 on RANKL-induced osteoclastogenesis in RAW264.7 cells.Methods: Vector or hIL-10 plasmid was transfected into 293T cells with liposome and thus hIL-10 protein was obtained.TRAP staining and bone absorption experiments were carried out to identify the biological function of hIL-10.Results: hIL-10 plasmid was transiently transfected into 293T and RAW264.7 cells successfully by liposome,with some cells emitting green fluorescence.ELISA detection confirmed that the target gene was expressed.RAW264.7 cells could be stably differentiated into osteoclast-like cells by RANKL.TRAP positive cells and bone resorption area significantly decreased(P<0.05) in the culture system,which was added with hIL-10 protein.Conclusion: The plasmids we used in this study could be transfected into 293T and RAW264.7 cells successfully by liposome,and the secreted hIL-10 protein had the biological function to inhibit osteoclast formation and bone resorption in vitro culture conditions.更多还原