抗纤维化短肽N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸对大鼠心脏成纤维细胞胶原合成与降解的调节作用

目的探讨N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸（AcSDKP）对血小板源性生长因子（PDGF）诱导的大鼠心脏成纤维细胞增殖、胶原合成和降解代谢的调节作用。方法分离培养新生大鼠心脏成纤维细胞。采用^3H-TdR和。H-脯氨酸掺入法分别检测心脏成纤维细胞增殖与胶原蛋白合成。Western blot法检测心脏成纤维细胞Ⅰ、Ⅲ型胶原蛋白表达和基质金属蛋白酶（MMP）-1蛋白的表达。明胶酶谱法检测心脏成纤维细胞MMP-2和MMP-9活性的表达。结果PDGF促进心脏成纤维细胞增殖、胶原合成,Ⅰ、Ⅲ型胶原表达,以及MMP-2、MMP-9活性和MMP-1表达。AcSDKP对PDGF介导的心脏成纤维细胞增殖、胶原合成均有抑制作用。AcSDKP上调由PDGF介导的心脏成纤维细胞MMP-2、MMP-9活性和MMP-1的表达。结论AcSDKP抑制PDGF介导的心脏成纤维细胞增殖和胶原的合成,上调MMPs活性或表达,促进胶原的降解,这些可能与AcSDKP抗心脏纤维化作用相关。

Role of AcSDKP on collagen synthesis and degradation in cultured rat cardiac fibroblast

OBJECTIVE: To investigate the role of AcSDKP on collagen synthesis and degradation in cultured rat cardiac fibroblasts. METHODS: Neonatal rat cardiac fibroblasts were isolated and stimulated by PDGF. The cell proliferation was observed by (3)H-TdR incorporation assay. The synthesis of collagen was measured by (3)H-proline incorporation assay. The expression of type I and type III collagen and MMP-1 protein were measured by Western blot. The MMP-2 and MMP-9 activity was evaluated with zymography assay. RESULTS: PDGF stimulated cardiac fibroblasts proliferation with increased collagen synthesis and type I and type III collagen protein expressions as well as MMP-2 and MMP-9 activities and MMP-1 expression. AcSDKP inhibited cardiac fibroblasts proliferation induced by PDGF and reduced collagen synthesis and type I and type III collagen protein expression. AcSDKP also further up-regulated MMP-2 and MMP-9 activities and MMP-1 expression in cardiac fibroblasts induced by PDGF. CONCLUSION: AcSDKP inhibited proliferation and collagen synthesis and up-regulated matrix metalloproteinases activity or expression induced by PDGF, which was possibly related with the effect of AcSDKP anti-fibrosis.