摘要
采用RNA干扰方法研究转录因子AP-1对TGF-β1的转录调控影响及其对心肌细胞的影响。设计合成两对AP-1基因的siRNA-59和siRNA-60,同时合成与AP-1基因序列无关的siRNA-58,试验设置阴性对照、阳性对照组及干扰组(AP-1-58、AP-1-59和AP-1-60)共5组,使用LipofectantineTM2000转染AP-1siRNA入培养的大鼠心肌细胞,倒置显微镜观察细胞形态;采用实时荧光定量PCR法检测AP-1和TGF-β1在心肌细胞中mRNA水平表达量的变化。结果显示,siRNA-58对AP-1的基因沉默(表达下调)无显著差异(P>0.05),siRNA-59和siRNA-60对AP-1的基因沉默(表达下调)有显著差异(P<0.05),同时对应siRNA-59和siRNA-60组的TGF-β1基因有显著性下调(P<0.05),说明AP-1和TGF-β1的基因表达存在正相关。
To study the effect of transcription factor AP-1 on TGF-β1 and cardiomyocytes by using RNA interference method,two pairs of siRNA of AP-1 genes were designed and synthesized,meanwhile unrelated siRNA of AP-1 genes was synthesized as control. The experiment was divided into five groups, including negative control group, positive control group and three interference groups（siRNA-58, siRNA-59, siRNA -60）. The cultured rat cardiomyocytes were transfected with AP-1 siRNA using LipofectamineTM2000. Cell morphology was observed by inverted microscope. The changes of mRNA expressions of AP-1 and TGF-131 in cardiomyocytes were detected by real-time quantitative PCR. The results showed, there was no significant difference in the expression of AP-1 gene in siRNA-58 group. In the siRNA-59 group and siRNA-60 group, the expression of AP-1 gene decreased significantly,while the expression of TGF-β1 gene was significantly reduced. The results indicated that the expression of AP-1 gene and TGF-β1 gene were positively correlated.
出处
《动物医学进展》
北大核心
2016年第12期46-49,共4页
Progress In Veterinary Medicine
基金
内蒙古自治区高等学校科学技术研究项目(NJZY12215)
内蒙古自然科学基金项目(2014MS0804
2016MS(LH)0826)
