摘要
应用RNA干扰技术建立大鼠肺泡巨噬细胞NR8383TGR5基因阻断模型。采用阳离子脂质体法,将TGR5siRNA(Santa Cruz Biotechnology)转染到NR8383细胞中,实时荧光定量PCR和Western blot分别检测NR8383细胞中TGR5mRNA和蛋白水平表达变化。结果显示,与对照组比,转染TGR5siRNA组可明显降低NR8383细胞TGR5mRNA和蛋白的表达(P<0.01)。RNA干扰技术能够特异性的高效阻断NR8383细胞中TGR5基因的表达,为深入探讨TGR5的生物学功能提供了可靠方法和新途径。
To establish a TGR5 receptor knockdown model of NR8383 rat alveolar macrophage with RNA interference technique,the TGR5 siRNA was transfected into NR8383 cells using the cationic liposome method,the expression changes of TGR5 mRNA and TGR5 protein in NR8383 cells were evaluated with qPCR and Western blot respectively.The results showed that,compared with normal control,the expres-sions of TGR5 mRNA and protein were significantly reduced by siRNA transfection in NR8383 cells （P 〈0.01）.RNA interference technique can effectively down regulate the expression of TGR5 gene in NR8383 cells specifically.This study provides a novel and reliable method fur further study on the biological func-tions of TGR5.
出处
《动物医学进展》
CSCD
北大核心
2014年第6期6-10,共5页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(31272605)
