Interference effect and significance of neonatal rat's lung fibroblast cells interfered by transforming growth factor β1 small interfering RNA

Objective To investigate the effect of transforming growth factor β1 (TGF-β1) small interfering RNA (siRNA) interference in transfecting neonatal rat's lung fibroblasts in vitro, and explore the best dose and maintenance time of TGF-β1 siRNA interference. Methods Neonatal rot' s lung fibroblasts cells were interfered with chemosynthesised TGF-β1 siRNA, effective siRNA was selected with real-time quantitative PCR,and then adenovirus vector for TGF-β1 siRNA was constructed to transfect neonatal rat' s lung fibroblasts. TGF-β1 mRNA expression was analyzed after transfection in vitro with real-time quantitative PCR to determine the best dose and maintenance time of TGF-β1siRNA interference. Results (1) After chemosynthesised siRNA interference for 24 h with 20 pmol, the second, third and forth siRNA all had different effect, and inhibition efficiency of the third siRNA was more than 70%. (2) TGF-β1 siRNA adenovirus vector was used to transfect neonatal rat's lung fibroblasts with 20 μl and 50 μl for 24 h, and then the inhibition effect was observed and compared with negative control group with 20 μl and blank group. The inhibition effect was obvious at 20 μl and 50 μl. The inhibition effect was observed with 20 μl for 24,48,72 h respectively, it showed that transfection for 24 h had strong inhibition effect, and inhibition effect of transfeotion increased more and more obviously for 48 h and 72 h. Conclusions The inhibition effect of TGF-β1 expression was obvious with TGF-β1 siRNA adenovirus vector in transfecting neonatal rat's lung fibroblast cells in vitro. Key words: RNA, small interfering;  Fibroblasts;  Transforming growth factor beta; Transfection;  Lung