Hepatocyte growth factor ameliorates myocardiac fibrosis by activating collagen degradation
Caiwen Ou, Guoqin Chen, Wenzhu Zhang, Zhijia Li, Minsheng Chen
Abstract
Caiwen Ou, Guoqin Chen, Wenzhu Zhang, Zhijia Li, Minsheng Chen
Abstract
Objective To study the effects of hepatocyte growth factor (HGF) on angiotensin Ⅱ(Ang Ⅱ )-induced synthesis and degradation of cardiac fibroblast collagen in neonatal mice.Methods Cardiac fibroblasts (CFs) of neonatal Sprague-Dawley (SD) rats were isolated by differential adhesion, and were identified by immunocytochemistry.The cells were assigned to normal control group (group C) , 10-6 mol/L Ang Ⅱ group (group A), 10μg/L HGF + 10-6 mol/L Ang Ⅱ group (group H1) and 100μg/L HGF +10-6 mol/L Ang Ⅱ group (group H2).All the groups were treated for 48 h.Hydroxyproline assay was used to determine the collagen protein level.RT-PCR was used to detect the expression of ColⅠ, ColⅢ, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-l) mRNA, and Western blotting was employed to measure Col I protein.In addition, MMP-1 activity was evaluated by collagenase Ⅰ activity assay kit.Results After intervention for 48 h, the collagen protein levels in groups A and H1 were significantly higher compared with group C [ (39.08±2.71) mg/L vs (37.45±4.22) mg/L vs (23.73 ±±1.62) mg/L, all P<0.05] and the collagen protein level in group H2 [(26.03±3.04) mg/L] was lower than those of groups A and Hl(P<0.05), but was not statistically different from group C.The expression levels of Col Ⅰ ,Col M and TIMP-1 showed an order of group A>group HI>group H2> group C (all P<0.05), and for MMP-1 mRNA, group A < group H1 < group H2 < group C (all P<0.05).The levels of Col I protein expression showed an order of group A>group HI>group H2>group C (89.90±4.29 vs 68.21?1.43 vs 36.08?.8 vs 30.14?.36, all P<0.05).The expression and activity of MMP-1 mRNA in group A and H1 were obviously lower than those in group C (all P<0.05), while group H2 were comparable to group C in these data.Conclusion HGF re-regulates MMP-1-TIMP-1 balance and ameliorates myocardiac fibrosis mainly by activating collagen degradation. Key words: Hepatocyte growth factor; Angiotensin E ; Matrix metalloproteinases; Cardiac fibroblasts; Collagen
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Objective To study the effects of hepatocyte growth factor (HGF) on angiotensin Ⅱ(Ang Ⅱ )-induced synthesis and degradation of cardiac fibroblast collagen in neonatal mice.Methods Cardiac fibroblasts (CFs) of neonatal Sprague-Dawley (SD) rats were isolated by differential adhesion, and were identified by immunocytochemistry.The cells were assigned to normal control group (group C) , 10-6 mol/L Ang Ⅱ group (group A), 10μg/L HGF + 10-6 mol/L Ang Ⅱ group (group H1) and 100μg/L HGF +10-6 mol/L Ang Ⅱ group (group H2).All the groups were treated for 48 h.Hydroxyproline assay was used to determine the collagen protein level.RT-PCR was used to detect the expression of ColⅠ, ColⅢ, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-l) mRNA, and Western blotting was employed to measure Col I protein.In addition, MMP-1 activity was evaluated by collagenase Ⅰ activity assay kit.Results After intervention for 48 h, the collagen protein levels in groups A and H1 were significantly higher compared with group C [ (39.08±2.71) mg/L vs (37.45±4.22) mg/L vs (23.73 ±±1.62) mg/L, all P<0.05] and the collagen protein level in group H2 [(26.03±3.04) mg/L] was lower than those of groups A and Hl(P<0.05), but was not statistically different from group C.The expression levels of Col Ⅰ ,Col M and TIMP-1 showed an order of group A>group HI>group H2> group C (all P<0.05), and for MMP-1 mRNA, group A < group H1 < group H2 < group C (all P<0.05).The levels of Col I protein expression showed an order of group A>group HI>group H2>group C (89.90±4.29 vs 68.21?1.43 vs 36.08?.8 vs 30.14?.36, all P<0.05).The expression and activity of MMP-1 mRNA in group A and H1 were obviously lower than those in group C (all P<0.05), while group H2 were comparable to group C in these data.Conclusion HGF re-regulates MMP-1-TIMP-1 balance and ameliorates myocardiac fibrosis mainly by activating collagen degradation. Key words: Hepatocyte growth factor; Angiotensin E ; Matrix metalloproteinases; Cardiac fibroblasts; Collagen
Key concepts: Hepatocyte growth factor, Endocrinology, Internal medicine, Collagenase, Fibroblast, Chemistry, Angiotensin II, Fibrosis