灯盏细辛对低氧环境下肺成纤维细胞Ⅰ型胶原、MMP1和TIMP1表达的干预作用

目的研究灯盏细辛对低氧环境下肺成纤维细胞Ⅰ型胶原、基质金属蛋白酶1(MMP1)和金属蛋白酶组织抑制剂1(TIMP1)表达的影响。方法以人胚肺成纤维细胞系MRC-5为研究对象,Western blot法检测常氧、低氧、丝/苏氨酸激酶抑制剂星型孢菌素(straurosporine,SP)、灯盏细辛作用下MRC-5缺氧诱导因子-1α(hypoxia-inducible factor-1alpha,HIF-1α)、Ⅰ型胶原前α1链(pro-col1α1)、p-smad2蛋白的表达,ELISA法检测细胞培养上清液中Ⅰ型胶原、MMP1和TIMP1蛋白的含量。结果体外培养的MRC-5,常氧环境下细胞内表达一定的pro-col1α1、p-smad2蛋白,几乎不表达HIF-1α,细胞外基质有较低水平的Ⅰ型胶原、MMP1和TIMP1蛋白表达;低氧环境下细胞内pro-col1α1、p-smad2蛋白、HIF-1α和细胞外基质Ⅰ型胶原、MMP1和TIMP1均较常氧组表达增加(P<0.05);SP(5nmol/L)预处理细胞30min与单纯低氧组比较,其胞内的p-smad2蛋白、pro-col1α1和基质中的Ⅰ型胶原、TIMP1降低(P<0.05),而基质中的MMP1表达增加(P<0.05);灯盏细辛组(12.5μg/mL、50μg/mL)与单纯低氧组比较其胞外的Ⅰ型胶原、MMP1、TIMP1蛋白和胞内的HIF-1α、pro-col1α1和p-smad2蛋白表达均有降低(P<0.05)。结论灯盏细辛可降低低氧环境下HIF-1α蛋白的表达,并部分通过p-smad2信号通路抑制Ⅰ型胶原、TIMP1蛋白生成。

Influence of Erigeron breviscapus on the expression of collagen type I , MMP1 and TIMP1 of MRC-5 cells under hypoxia

Objective Study the influence of erigeron breviscapus on collagen typeⅠ,matrix metalloproteinase1(MMP1) and tissue inhibitor of metalloproteinase1(TIMP1) in human embryo lung fibroblast MRC-5 under hypoxia.Methods MRC-5 cultured in vitro were individed into 4 groups,including normoxia control,hypoxia,hypoxia and staurosporine(SP),hypoxia and erigeron breviscapus.The expression of hypoxia-inducible factor-1α(HIF-1α),pro-col1α1 and phospho-smad2(p-smad2) were evaluated by Western blot.The level of collagen typeⅠ,MMP1 and TIMP1 in supernatant fluid were detected by ELISA.Results Expressed pro-col1α1,p-smad2 in MRC-5 cells under nomoxia were detected,but not HIF-1α.A low level of collagen typeⅠ,MMP1 and TIMP1 was also detected in the cell supernatant.In nomoxia secreted pro-col1α1,p-smad2,collagen typeⅠ,MMP1 and TIMP1,hardly secreted HIF-1α.Under hypoxia,the expression of all of them was increased when compared with those of under nomoxia condition(P<0.05).MMP1 expression was significantly further up-regluted by pre-cubation with SP in hypoxia,however collagen typeⅠ,TIMP1 in supernatant fluid and pro-col1α1,p-smad2 in cells were inhibited(P<0.05,vs.hypoxia group).Treated with Erigeron breviscapus(12.5 μg/mL,50 μg/mL) under hypoxia condition,significantly reduced expression of pro-col1α1,p-smad2,HIF-1α in MRC-5 cells and collagen typeⅠ,MMP1,TIMP1 in supernatant fluid were observed(P<0.05,vs.hypoxia).Conclusion Erigeron breviscapus can decrase the expression of HIF-1α of MRC-5 cells under hypoxia condition by inhibit the expression of collagen typeⅠ and TIMP1 partly through the Smad2 signaling pathways.