Objective To investigate the effects of Smad7 knock down by
lentivirus on
rat cardiac
fibroblasts proliferation,
migration, cell differentiation and
collagen secretion in vitro.
Methods The primary cardiac
fibroblasts were
separated from the
hearts of ten SD
rats and identified by immunohistochemical
method. The
lentivirus transfection knocked down the expresson of Smad7 in cardiac
fibroblasts,
Western blotting was used to detect the
efficiency of Smad7 knock down by
lentivirus. The proliferation of cardiac
fibroblasts was quantified by real-
time unlabeled
cell analyzer.
Cell migration was evaluted by
cell wound scratch assay.
Western blotting was used to detect expression of α-
smooth muscle actin(α-SMA) and
collagen Ⅰ(Col Ⅰ). Results Myocardial
fibroblasts were successfully cultured and identified by immunocytochemical
methods. The multiplicity of
infection(MOI) that
lentivirus transduction of myocardial
fibroblasts was 100. After
lentivirus transduction, 88.33% myocardial
fibroblasts expressed
green fluorescent protein, showed that the
lentivirus could significantly reduce the
protein expression of Smad7. Smad7
deficiency decreased the proliferation and migration of cardiac
fibroblasts, increased the
protein expression of α-SMA and decreased
collagen secretion. The results indicated that Smad7
deficiency significantly down-regulated the proliferation and migration of cardiac
fibroblasts, increased α-SMA
protein expression and reduced ColⅠ
protein expression. Conclusion Smad7
deficiency can significantly change the cardiac
fibroblasts function, that is related to the pathological mechanism that
lead to myocardial
fibrosis