大鼠视网膜中转化生长因子β1和β2基因表达的定量检测

目的：定量检测转化生长因子-β1（Transfimning growth factorβ1，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保持宅好未被降解，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．前者明显高干后者，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的比值由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在视网模中以TGF-β2表达为主，提示可能是TGF-β2在视网膜病变中起主导怍用。

Quantitative detection of the expression level of transforming growth factor-β1 and -β2 in rat retina with real time PCR

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.