蜈蚣酸性蛋白对血管紧张素-Ⅱ诱导心肌成纤维细胞增殖和胶原合成的影响

Effects of centipede acidic protein on the proliferation and collagen synthesis of cultured neonatal rat cardiac fibroblast induced by angiotensinⅡ

目的:研究蜈蚣酸性蛋白(centipede acidic protein,CAP)对血管紧张素-Ⅱ(AngⅡ)诱导培养心肌成纤维细胞(cardiac fibroblasts,CFb)增殖和胶原合成的影响,探讨其抑制心肌纤维化的机制。方法:建立AngⅡ诱导乳鼠CFb纤维化模型,噻唑蓝(MTT)法检测CAP对CFb增殖的影响;羟脯氨酸测定CFb胶原合成量;硝酸还原酶法测定细胞培养液中NO含量;半定量逆转录聚合酶链反应(RT-PCR)法测定相关基因c-myc mR-NA表达。结果:AngⅡ模型组与空白对照组比较,CFb增殖及胶原合成显著增加、NO含量显著降低、c-mycmRNA的表达显著增加(P<0.01);CAP高、低剂量组与AngⅡ模型组比较,CFb增殖及胶原合成显著降低、NO含量显著升高、c-myc mRNA的表达显著下调(P<0.05或P<0.01)。结论:CAP具有明显的抑制心肌纤维化作用,其机制与提高NO含量及下调c-myc mRNA表达有关。

Objective： To study the influence of Centipede Acidic Protein （CAP） on the proliferation and collagen synthesis of cultured neonatal rat cardiac fibroblasts （CFb） induced by angiotensin Ⅱ （Ang Ⅱ）, and to explore the mechanisms of CAP on cardiac fibrosis. Methods： Neonatal rat cardiac fibroblasts were treated with Ang Ⅱ to produce fibrosis model. The effects of CAP on proliferation of CFb were observed by MTT colorimetric assay, syn- thesis of collagen was observed by the hydroxyproline concentration. The NO contents were measured by Nitric acid reductase method. The c-myc expression was examined by semi-quantitative RT-PCR analysis. Results： Compared with that of control group, the proliferation, collagen synthesis and the levels of c-myc mRNA expression of CFb in the model group increased, while the NO contents decreased obviously（ P 〈 0.01 ）. Compared with that of model group, the proliferation, collagen synthesis and the levels of c-myc mRNA expression of CFb in the CAP groups decreased, while the NO contents increased obviously（ P 〈 0.05 or P 〈 0.01 ）. Conclusion ： The results suggested that CAP significantly inhibited cardiac fibrosis and its mechanism may be increase the NO contents and down-regulate c- myc mRNA expression.