西红花酸在AngⅡ诱导的大鼠肺成纤维细胞中的作用

Anti-fibrotic Effects of Crocetin on Lung Fibroblasts Induced by AngⅡ

目的:通过观察西红花酸(CRT)对血管紧张素Ⅱ(AngⅡ)诱导的肺成纤维细胞(LFB)增殖及转化生长因子β1(TGF-β1)、Ⅰ型胶原(COL-Ⅰ)、α-平滑肌肌动蛋白(α-SMA)、ERK1/2和P-ERK1/2基因蛋白水平的影响,探讨其抗肺纤维化的作用机制。方法:酶消化法提取原代LFB,采用四氮唑盐(MTT)比色法测定细胞增殖,RT-PCR法相对定量检测TGF-β1和COL-ⅠmRNA表达,Western Blot法检测α-SMA、ERK1/2和P-ERK1/2蛋白的表达。结果:10-7、10-6 mol.L-1西红花酸可显著减少AngⅡ诱导的LFB细胞增殖,降低TGF-β1和COL-ⅠmRNA的表达,降低α-SMA和P-ERK1/2的蛋白表达。结论:西红花酸可显著减少AngⅡ诱导的LFB细胞增殖,并呈剂量依赖关系,其作用机制可能与调节TGF-β1-ERK1/2信号通路有关。

Objective：To investigate the effects of crocetin on angiotensin Ⅱ（Ang Ⅱ）-induced proliferation,collagen synthesis and α-smooth muscle actin（α-SMA） in rat lung fibroblast（LFB） cells,and the anti-fibrosis mechanism.Methods：LFB cells were treated with crocetin at various concentrations for 24 h or 48 h in the absence or presence of Ang Ⅱ.Cell proliferation was determined by methyl thiazolyl tetrazolium（MTT） reduction assay,the expression levels of transforming growth factor-β1（TGF-β1） mRNA and type Ⅰ collagen（COL-Ⅰ） mRNA were assayed by real time-PCR.The expression of α-SMA,extracellular-regulated kinase 1/2（ERK 1/2） and phosphorylation of extracellular-regulated kinase 1/2（P-ERK 1/2） were measured by Western blot.Results：Crocetin inhibited Ang Ⅱ-induced LFB cell proliferation,and could markedly suppress the up-regulation of TGF-β1 mRNA,COL-Ⅰ mRNA,α-SMA and P-ERK1/2（P0.05 or P0.01） in a concentration-dependent manner.Conclusion：These results indicate that crocetin inhibits Ang Ⅱ-induced fibrosis,at least partly,through TGF-β1-ERK1/2 signal pathway.