补阳还五汤对缺氧心脏成纤维细胞旁分泌MMP-2/9、TIMP-1、TNF-α的影响

Effects of Buyang Huanwu decoction on the expression of MMP-2/9, TIMP-1, TNF-α in hypoxia induced cardiac fibroblasts

关键词:  补阳还五汤  心脏成纤维细胞  缺氧  MMP-2  MMP-9  TIMP-1  TNF-α

Key words:  Buyang Huanwu decoction  cardiac fibroblast  hypoxia  MMP-2  MMP-9  TIMP-1  TNF-α

摘要:

[目的] 探讨补阳还五汤对缺氧诱导心脏成纤维细胞旁分泌基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶组织抑制因子1(TIMP-1)及肿瘤坏死因子-α(TNF-α)的影响,并从基因和蛋白水平探讨其保护心肌细胞、预防心肌纤维化的机制。[方法] 培养小鼠乳鼠心脏成纤维细胞,将其分为对照组、模型组、补阳还五汤组。采用逆转录聚合酶链式反应(RT-PCR)法和酶联免疫吸附测定(ELISA)法分别检测各组成纤维细胞和细胞培养上清液中MMP-2、MMP-9、TIMP-1及TNF-α mRNA表达和蛋白分泌。[结果] 与对照组比较,模型组MMP-2、MMP-9及TNF-α mRNA表达上调,蛋白旁分泌增加,TIMP-1表达下调(P<0.05);与模型组比较,补阳还五汤组MMP-2、MMP-9 mRNA表达下调,蛋白旁分泌减少,TIMP-1则上调(P<0.05),而TNF-α仅有下调趋势,无统计学差异(P>0.05)。[结论] 补阳还五汤可保护缺氧心肌,预防心肌纤维化。其机制可能与抑制缺氧成纤维细胞的旁分泌,调节MMP-2、MMP-9、TIMP-1的表达平衡有关。

Abstract:

[Objective] To observe the effect of Buyang Huanwu decoction on the paracrine of cardiac fibroblasts MMP-2, MMP-9, TIMP-1 and TNF-α in mice with hypoxia and explore the molecular mechanism in protecting myocardial cells. [Methods] Culture cardiac fibroblasts of neonatal mice, divided into three groups: control group, model group, traditional Chinese medicine (TCM) group, copied the anoxic model in condition of 5% CO₂ + 94% N₂ + 1% O2 for 24 h. MMP-2, MMP-9, TIMP-1 and TNF-α mRNA and protein expression were detected by RT-PCR and ELISA. [Results] Compared to control group, the mRNA and protein expression of MMP-2, MMP-9 and TNF-α in model group were increased, while the TIMP-1's expression were induced. After Buyang Huanwu decoction intervention, the MMP-2, MMP-9 and TNF-α in TCM group were decreased, and the TIMP-1 was increased (P<0.05). While compared to model group TNF-α had no significant difference (P>0.05). [Conclusion] Buyang Huanwu decoction can reduce the expression of MMP-2, MMP-9 and increase TIMP-1 in hypoxic cardiac fibroblasts to protect myocardial cells, prevent and relieve the development of myofibrosis.