Effects of Notch1 on TGF-β1 induced transdifierentiation of cardiac fibroblasts to myofibroblast
AIM: To investigate the change of Notchl in the transdifferentiation of cardiac fibroblasts ( CFs ) into myofibroblasts (MFB) induced by TGF-β_1 and the possible effects of Notchl during this progess. METHODS: CFs were obtained and cultured in vitro, and were stimulated with TGF-β_1 ( 10 μg/L)to promote the transdifferentiation of CFs into MFB. The CFs were divided into control group, 24, 48 and 72 h group based on the time length of TGF-β_1 stimulation. Hydroxyp roline (HYP) content was detected by alkaline hydrolysis colorimetry, the expression level of a-smooth muscle actin(α-SMA) protein was measured by Western Blot and immunofluorescence, and the expression levels of Notch mRNA and protein were detected by quantitative real-time PCR and Western Blot. Gamma-secretase inhibitor, N-[ N-( 3,5-diflu-orophenacetyl) -l-alanyl ] -S-phenylglycine t-butyl ester, DAPT ( 75Ixmol/L) was then used to block the activation of Notehl receptors,and the CFs were divided into control group, 24, 48 and 72 h group based on the time length of DAPT stimulation. Then the expression levels of α-SMA and HYP were detected. RESULTS: As the time extension of TGF-β_1 stimulation, the expression levels of HYP and α-SMA increased gradually while Notchl declined compared with the control group, with the most significant effect in 72 h group(P <0.01 ). As the incubating time with DAPT prolonging, the expression levels of ot-SMA and HYP rose compared with the control group, with the most significant effect in 72 h group(P <0.01). CONCLUSION: TGF-β_l can induce the transdifferentiation of CFs into MFB, in which the expression level of Notchl decreased. DAPT can block the activation of Notchl and promote the transdifferentiation. Thus, Notchl may play an important role in the transdifferentiation of CFs into MFB, and it will possibly become a new therapeutic target for blocking even reversing the cardiac fibrosis.
NETL
NSTL
万方数据
维普
