贝前列素对心肌成纤维细胞胶原交联的影响及机制研究

Effects and mechanisms of beraprost on cross linked collagen in cardiac fibroblasts

目的 :观察贝前列素对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)刺激下心肌成纤维细胞胶原交联的影响并初步探讨其机制。方法:体外培养新生大鼠心肌成纤维细胞,给予贝前列素预处理后,AngⅡ(100 nmol/L)再刺激24 h,测定细胞培养基中胶原含量和交联程度,实时荧光定量PCR和Western blot法测定赖氨酰氧化酶(lysyl oxidase,LOX)m RNA和蛋白表达。结果:贝前列素可剂量依赖性地抑制AngⅡ刺激下心肌成纤维细胞胶原分泌,其中10μmol/L作用最强。贝前列素(10μmol/L)预处理4 h后再给予AngⅡ刺激,心肌成纤维细胞胶原分泌明显减少,交联胶原水平下降,交联与非交联胶原比例变低;LOX m RNA和蛋白表达亦明显下降。结论:贝前列素抑制AngⅡ刺激下心肌成纤维细胞胶原交联,机制可能与下调LOX表达有关。

Objective：To observe the effects and mechanisms of beraprost on cross linked collagen in angiotensin Ⅱ（Ang Ⅱ ）- stimulated cardiac fibroblasts. Methods：The cultured neonatal rat cardiac fibroblasts were pre-treated with beraprost followed with Ang Ⅱ （100 nmol/L） stimulation for 24 h. Total collagen and cross linked collagen were examined in culture medium. Lysyl oxidase （LOX） expression was determined with real time PCR and western blot. Results：Beraprost dose-dependently inhibited collagen secretion with Ang [I stimulation,among which 10 μmol/L presented with the most robust suppression. Beraprost （10 μmol/L） pre-treatment for 4 h followed with Ang 11 stimulation significantly reduced collagen secretion,as well as the cross linked degree and ratio of cross linked collagen to non-cross linked collagen, accompaying with decreasing mRNA and protein expression of LOX. Conclusion： Beraprost inhibited cross linked collagen in Ang 11-stimulated cardiac fibrohlasts,which may be associated with suppression of L0X expression.