Egr-1诱骗寡核苷酸下调PDGF抑制原代培养的血管平滑肌细胞增殖
发布时间：2018-11-21 12:59
【摘要】： Egr-1诱骗寡核苷酸下调PDGF—A抑制原代培养的血管平滑肌细胞增殖 目的 通过向原代培养的大鼠动脉平滑肌细胞中转染针对早期生长反应因子-1(early growth response factor-1,Egr-1)的诱骗寡核苷酸(decoy oligonucleotide,decoy ODN),检测血管平滑肌细胞增殖情况及早期生长反应因子-1、血小板衍生生长因子-A(platelet derived growth factor,PDGF-A)的表达变化,探讨针对早期生长反应因子-1的诱骗寡核苷酸对体外培养的大鼠平滑肌细胞增殖的影响和作用机制。 方法 通过组织块贴壁法进行原代大鼠动脉血管平滑肌细胞(Vascular smoorh muscle cells VSMC)培养、传代培养和鉴定,设计合成Egr-1诱骗、诱骗对照寡核苷酸.转染人工合成的针对Egr-1的decoy ODN,将培养的平滑肌细胞分为对照组(正常培养细胞)、诱骗组(转染Egr-1的诱骗寡核苷酸)、诱骗对照组(转染Egr-1诱骗杂码寡核苷酸).免疫细胞化学染色法检测转染前后平滑肌细胞增殖情况与Egr-1及增殖细胞PDGF-A的表达变化,进行图像分析。所有数据均采用SAS8.0统计软件进行统计分析,计量资料以均值±标准差(x±S)表示,各组间比较应用单因素方差分析,P0.05有统计学意义。 结果 成功进行原代VSMC培养,经细胞鉴定培养细胞为高纯度的血管平滑肌细胞。转染Egr-1诱骗寡核苷酸后,用免疫组织化学法分别观察各组不同时间点的Egr-1和PDGF表达的情况发现：Egr-1在三个实验组中均有表达,诱骗转染组与诱骗对照组和阴性对照组相比,均在一定程度上抑制了Egr-1和PDGF-A表达,经SAS8.0统计软件进行统计分析,证明有统计学意义(P0.05)。 结论 本实验证实Egr-1是一种重要的核转录因子,体外转染针对Egr-1的诱骗寡核苷酸在一定程度上能够抑制血管平滑肌细胞Egr-1及PDGF-A的表达,从而达到抑制细胞增殖的目的,在抗动脉粥样硬化和治疗血管再狭窄方面具有重要作用,从而为基因治疗再狭窄、防治动脉粥样硬化开辟了一条新的途径。
[Abstract]:Egr-1 decoy oligodeoxynucleotides down-regulate the proliferation of primary cultured vascular smooth muscle cells by down-regulation of PDGF-A by transfection into primary cultured rat arterial smooth muscle cells for early growth response Factor-1 (early growth response factor-1, Egr-1 was used to detect the proliferation of vascular smooth muscle cells and the expression of early growth response factor-1 and platelet-derived growth factor A (platelet derived growth factor,PDGF-A. To investigate the effect and mechanism of decoy oligonucleotides targeting early growth response factor-1 on the proliferation of rat smooth muscle cells in vitro. Methods the primary vascular smooth muscle cells (Vascular smoorh muscle cells VSMC) of rat artery were cultured, subcultured and identified by tissue block adherence method. Egr-1 decoy was designed and synthesized, and the control oligonucleotides were decoded. The cultured smooth muscle cells were divided into control group (normal cultured cells) and decoy group (decoy oligonucleotides transfected with Egr-1). Decoy control group (transfection of Egr-1 decoy code oligonucleotide). The proliferation of smooth muscle cells and the expression of Egr-1 and PDGF-A were detected by immunocytochemical staining before and after transfection. All the data were analyzed by SAS8.0 statistical software. The measurement data were expressed as mean 卤standard deviation (x 卤S). Results the primary VSMC was successfully cultured and the cultured cells were identified as high purity vascular smooth muscle cells. After transfection of Egr-1 to induce oligonucleotides, the expression of Egr-1 and PDGF at different time points in each group were observed by immunohistochemical method. The results showed that Egr-1 was expressed in all three experimental groups. The expression of Egr-1 and PDGF-A was inhibited to some extent in the decoy transfection group compared with the decoy control group and the negative control group. The statistical analysis by SAS8.0 software proved that there was statistical significance (P0.05). Conclusion Egr-1 is an important nuclear transcription factor. In vitro transfection of decoy oligonucleotides against Egr-1 can inhibit the expression of Egr-1 and PDGF-A in vascular smooth muscle cells to some extent. Thus, it can inhibit cell proliferation and play an important role in anti-atherosclerosis and vascular restenosis, thus opening a new way for gene therapy and prevention and treatment of atherosclerosis.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363
本文编号：2347018
[Abstract]:Egr-1 decoy oligodeoxynucleotides down-regulate the proliferation of primary cultured vascular smooth muscle cells by down-regulation of PDGF-A by transfection into primary cultured rat arterial smooth muscle cells for early growth response Factor-1 (early growth response factor-1, Egr-1 was used to detect the proliferation of vascular smooth muscle cells and the expression of early growth response factor-1 and platelet-derived growth factor A (platelet derived growth factor,PDGF-A. To investigate the effect and mechanism of decoy oligonucleotides targeting early growth response factor-1 on the proliferation of rat smooth muscle cells in vitro. Methods the primary vascular smooth muscle cells (Vascular smoorh muscle cells VSMC) of rat artery were cultured, subcultured and identified by tissue block adherence method. Egr-1 decoy was designed and synthesized, and the control oligonucleotides were decoded. The cultured smooth muscle cells were divided into control group (normal cultured cells) and decoy group (decoy oligonucleotides transfected with Egr-1). Decoy control group (transfection of Egr-1 decoy code oligonucleotide). The proliferation of smooth muscle cells and the expression of Egr-1 and PDGF-A were detected by immunocytochemical staining before and after transfection. All the data were analyzed by SAS8.0 statistical software. The measurement data were expressed as mean 卤standard deviation (x 卤S). Results the primary VSMC was successfully cultured and the cultured cells were identified as high purity vascular smooth muscle cells. After transfection of Egr-1 to induce oligonucleotides, the expression of Egr-1 and PDGF at different time points in each group were observed by immunohistochemical method. The results showed that Egr-1 was expressed in all three experimental groups. The expression of Egr-1 and PDGF-A was inhibited to some extent in the decoy transfection group compared with the decoy control group and the negative control group. The statistical analysis by SAS8.0 software proved that there was statistical significance (P0.05). Conclusion Egr-1 is an important nuclear transcription factor. In vitro transfection of decoy oligonucleotides against Egr-1 can inhibit the expression of Egr-1 and PDGF-A in vascular smooth muscle cells to some extent. Thus, it can inhibit cell proliferation and play an important role in anti-atherosclerosis and vascular restenosis, thus opening a new way for gene therapy and prevention and treatment of atherosclerosis.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363
【参考文献】
相关期刊论文 前4条
1 段佑才,姜泊,智发朝,宋于刚,陈学清;Egr-1基因剔除对雨蛙素诱导小鼠胰腺炎的影响[J];第四军医大学学报;2005年16期
2 金蓉,许志成,刘阳,刘伟,崔磊,曹谊林;体外不同培养方法对血管平滑肌细胞生物学性状影响的比较研究[J];实用美容整形外科杂志;2003年04期
3 鄢定红,程晓曙,王雪梅,张润香,李菊香,苏海,吴清华;一氧化氮缺乏在高血压及心血管重构中的作用[J];高血压杂志;2000年04期
4 俞建华,程晓曙,杨人强,刘东,苏海,吴清华;他汀对NO缺乏性高血压大鼠主动脉重构的作用[J];高血压杂志;2002年06期
，本文编号：2347018
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2347018.html
最近更新
教材专著
