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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 effect of beraprost on collagen crosslinking of cardiac fibroblasts stimulated by angiotensin Ⅱ (AngⅡ) and to explore its mechanism. Methods: The neonatal rat cardiac fibroblasts were cultured in vitro. After beraprost preconditioning, Ang Ⅱ (100 nmol / L) was restimulated for 24 h. The contents of collagen and crosslinking in the cell culture medium were determined. Real-time PCR and Western blot was used to determine the mRNA and protein expression of lysyl oxidase (LOX). Results: Beraprost inhibited the collagen secretion of cardiac fibroblasts in a dose-dependent manner. The effect of beraprost was the strongest at 10μmol / L. After pre-treatment with beraprost (10 μmol / L) for 4 h, the collagen secretion of cardiac fibroblasts was significantly decreased and AngⅡ was decreased. The level of crosslinked collagen decreased and the ratio of crosslinked and uncrosslinked collagen decreased. LOX mRNA and protein expression Also significantly decreased. CONCLUSION: Beraprost inhibits the collagen cross-linking of cardiac fibroblasts induced by AngⅡ, which may be related to the down-regulation of LOX expression.