香菇多糖对白介素-1β诱导人胚肺成纤维细胞表型转化的影响

Effects of lentinan on interleukin-1β-induced transdifferentiation of human embryonic lung fibroblasts to myofibroblasts

摘要:

目的：研究白介素-1β（IL-1β）对人胚肺成纤维细胞(FB)向肺肌成纤维细胞转化的影响及香菇多糖（LNT）的作用。方法：体外培养人胚肺FB，用不同浓度IL-1β和LNT作用于细胞后，采用CCK-8法检测细胞的增殖情况，免疫细胞化学染色检测α-平滑肌肌动蛋白（α-SMA），RT-PCR检测纤维连接蛋白（FN）、Ⅰ型胶原蛋白（ColⅠ）和α-SAM mRNA的相对表达量。结果：①IL-1β组细胞增殖的吸光度、α-SMA蛋白及FN、ColⅠ和α-SAM mRNA表达均高于对照组（P<0.01），且随着IL-1β浓度的增加，表达逐渐增高。②LNT呈浓度依赖性抑制IL-1β诱导的细胞增殖及α-SMA蛋白、FN、ColⅠ和α-SAM mRNA表达（P<0.01）。结论：LNT可抑制IL-1β诱导人胚肺FB增殖和肌成纤维细胞转化及细胞外基质积聚。

Abstract:

OBJECTIVE: To study the effects of interleukin-1β (IL-1β) on transdifferentiation of human embryonic lung fibroblasts to myofibroblasts and the effects of lentinan on the transdifferentiation. METHODS: The human embryonic lung fibroblasts were cultured in vitro, and fibroblasts were treated with different concentrations of IL-1β and lentinan. The proliferation activity of the human embryonic lung fibroblasts was evaluated by the Cell Counting Kit-8 (CCK-8). The expression of α-smooth muscle actin (α-SMA) protein was measured by immunocytochemistry. The levels of fibronectin (FN), typeⅠcollagen (ColⅠ) and α-SMA mRNA were detected by RT-PCR. RESULTS: Compared with the untreated control group, the absorbance value of cell proliferation, α-SMA protein levels, FN, ColⅠand α-SMA mRNA expression were significantly up-regulated after different concentrations of IL-1β (0.1, 1, 10 ng/mL) treatment for 48 hrs (P<0.01). Lentinan treatment inhibited up-regulation of the cell proliferation activity, α-SMA protein levels, FN, ColⅠand α-SMA mRNA expression induced by IL-1β in a dose-independent manner (P<0.01). CONCLUSIONS: Lentinan can suppress human embryonic lung fibroblast proliferation, fibroblast-myofibroblast transdifferentiation and extra cellular matrix synthesis induced by IL-1β.