N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸对转化生长因子β1介导的心脏成纤维细胞胶原代谢的调节作用

Regulation effect of AcSDKP on cardiac fibroblasts and collagen metabolism mediated by TGF-β1 in rats

目的探讨N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸（AcSDKP）对转化生长因子-β1（TGF-β1）介导的大鼠心脏成纤维细胞增殖与胶原代谢的调节作用。方法采用差速贴壁法获取新生大鼠心脏成纤维细胞；3H—TdR掺入法检测心脏成纤维细胞的增殖，免疫细胞化学染色法检测增殖细胞核抗原（PCNA）的表达。3H-脯氨酸掺入法检测心脏成纤维细胞胶原蛋白的合成，Westernblot法检测Ⅰ、Ⅲ型胶原蛋白与MMP-1、TIMP-1蛋白的表达。结果AcSDKP抑制了TGF—β1对心脏成纤维细胞MMP-1蛋白表达的下调作用，使MMP-1蛋白表达增加，而对TGF—β1介导的TIMP-1蛋白表达无明显影响，使MMP-1／TIMP-1比值增加。结论AcSDKP抑制TGF—β1介导的心脏成纤维细胞的增殖、胶原合成，上调MMP-1蛋白表达，使MMP-1／TIMP-1比值增加，这可能与AcSDKP抗心脏纤维化的作用相关。

Objective To investigate the regulation effect of N-acetyl-seryl-aspartyl-lysyl-proline （AcSDKP） on the proliferation of cardiac fibroblasts and collagen metabolism mediated by transforming growth factor-β1 （TGF-β1） in rats. Methods The rat cardiac fibroblasts were isolated by applying differential adhesion method. The proliferation of cardiac fibroblasts were examined by using 3 H-TdR assay, and the expression of proliferating cell nuclear antigen （PCNA） was detected with immunocytochemistry. The collagen synthesis of cardiac fibroblasts was examined by using 3 H-proline incorporation assay, and the expressions of collage Ⅰ ,collage Ⅲ ,MMP-1 and TIMP-1 were detected with Western blot method. Results TGF-β1 promoted the proliferation of cardiac fibroblasts and collagen synthesis, increased the expressions of collage Ⅰ and collage Ⅲ ,decreased MMP-1 expression,improved TIMP-1 expression, and reduced the ratio of MMP-1 to TIMP-1. AcSDKP had the inhibitory effect on the proliferation of cardiac fibroblasts and collagen synthesis mediated by TGF-β1. AcSDKP inhibited too the down-regulation of TGF-β1 on MMP-1 expression, and increased MMP-1 expression. AcSDKP had no influence on TIMP-1 expression, but improved the ratio of MMP- 1 to TIMP-1. Conclusion AcSDKP can inhibit the proliferation of cardiac fibroblasts and collagen synthesis mediated by TGF-β1, ,up-regulate MMP-1 expression,and increase the ratio of MMP-1 to TIMP-1 ,which may be related to the anti-fibrosis effect of AcSDKP.