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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。
Objective: To quantitatively detect the expression of transforming growth factor β1 (TGF-β1) and transforming growth factor-β2 (TGF-β2) in the normal retina of rats and to explore the role of TGF-β1 and TGF-β2 in the retina Differences in Expression and Their Significance. Methods: The normal retina was isolated and extracted from rat retina. RNA was extracted and reverse transcribed. The mRNA levels of TGF-β1 and TGF-β2 were analyzed by real-time fluorescence quantitative PCR. RESULTS: Rat retinal RNA remained intact without degradation and could be used for expression profiling. The gene expression level of TGF-β2 relative to β-actin in the rat retina was 0.0378 ± 0.009 and that of TGF-β1 was 0.0008 ± 0.0003, the former was significantly higher than the latter (t = 12.37, P < 0.001), indicating that the expression of TGF-β in the retina mainly TGF-β2, TGF-β2 and TGF-β1 ratio of 55.00 ± 26.61. Conclusion: Real-time fluorescence quantitative PCR can accurately and precisely analyze the gene expression of very few tissue cells. TGF-β in the retina to TGF-β2-based expression, suggesting that TGF-β2 may play a leading role in retinopathy.