摘要
目的:定量检测转化生长因子-β1(Transforming growth factor β1,TGF-β1)和转化生长因子-β2(TGF-β2)在大鼠正常视网膜中的表达水平,探讨TGF-β1和TGF-β2在视网膜中表达的差异及其意义。方法:分离取出大鼠正常视网膜,抽提RNA并逆转录,实时荧光定量PCR技术分析TGF-β1和TGF-β2的mRNA含量。结果:大鼠视网膜RNA保持完好未被降解,能够用于表达水平分析。大鼠视网膜中TGF-β2相对于β-actin的基因表达水平为0.0378±0.009,TGF-β1为0.0008±0.0003,前者明显高于后者,统计学上差异有显著性(t=12.37,P<0.001),说明在视网膜中TGF-β的表达以TGF-β2为主,TGF-β2和TGF-β1的比值为55.00±26.61。结论:实时荧光定量PCR技术能够针对性地精确分析极少量组织细胞的基因表达。TGF-β在视网膜中以TGF-β2表达为主,提示可能是TGF-β2在视网膜病变中起主导作用。
AIM- To quantitatively detect the expression level of transforming growth factor-β1（TGF-β1） and transforming growth factor-β2 （TGF-β2） genes in the retina of normal rat in order to determine the expression difference of TGF-β1 and TGF-β2 in retina.
METHODS： The total RNA was isolated from which the first strand of cDNA was prepared. The mRNA levels of TGF-β1 and TGF-β2 were detected quantitatively by real time polymerase chain reaction （PCR）.
RESULTS： The mRNA levels of TGF--β1 and TGF-β2 were 0. 0008± 0. 0003 and 0. 0378±0. 009, respectively. Expression of TGF-β2 was obviously higher than that of TGF-β1 in rat retina with statistical significance （ t= 12.37, P 〈 0. 001 ）. The ratio of TGF-β2/TGF-β1 was 55.00± 26.61.
CONCLUSION： QRT-PCR could specifically and accurately detect gene expression level in rat retina. In retina the TGF-β2 gene was expressed more abundantly than TGF-β1. It is suggested that TGF-β2 play an important role in retina diseases.
出处
《国际眼科杂志》
CAS
2008年第6期1076-1078,共3页
International Eye Science
基金
中国国家自然科学基金资助项目(No.30271391)
中国上海市卫生局科研项目(No.034124)
中国上海市卫生系统百名跨世纪优秀学科带头人培养计划(百人计划)基金资助项目(No.057)~~
关键词
视网膜
实时荧光定量PCR
转化生长因子-Β
基因表达
retina
quantitative reverse transcription polymerase chain reaction （QRT-PCR）
TGF-β
gene expression
