南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (10): 1411-.

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IL-27及其受体对肺成纤维细胞增殖及转化的影响

董昭兴,邰文琳,雷雯,王颖,李振坤,张涛   

  • 出版日期:2015-10-20 发布日期:2015-10-20

Effects of interleukin-27 and its receptor on TGF-β1-induced murine pulmonary fibroblast
proliferation and transformation

  • Online:2015-10-20 Published:2015-10-20

摘要: 目的探讨IL-27及其受体(WSX-1)对TGF-β1诱导的小鼠肺成纤维细胞增殖、转化及胶原合成作用。方法构建IL-27R/
pCDNA3.1 过表达载体。实验分为6 个组:正常组,TGF-β1 组,IL-27 组,IL-27 受体组,IL-27+IL-27 受体组,TGF-β1+IL-27+
IL-27 受体组。光学显微镜下观察细胞的生长状况,用MTT检测各处理组对肺成纤维细胞增殖的影响。RT-PCR 和Western
blotting 检测肌成纤维细胞标志物a-SMA以及Ⅰ和Ⅲ型胶原的表达,并用免疫荧光检测a-SMA蛋白的定位及表达情况。结果
(1)IL-27和IL-27R都可以抑制肺成纤维细胞增殖;(2)TGF-β1促进肺成纤维细胞表达α-SMA增加,并促进肺成纤维细胞Ⅰ和
Ⅲ型胶原合成;(3)IL-27及其受体可以抑制肺成纤维细胞α-SMA的表达,并减少肺成纤维细胞I和III型胶原的合成;(4)IL-27/
IL-27R共同作用时,对肺成纤维细胞增殖转化没有影响。结论IL-27或IL-27受体可以抑制TGF-β1诱导的小鼠肺纤维化细胞增
殖、转化和胶原的合成作用,但是IL-27与IL-27受体共同存在的情况下却不能发挥抑制作用,这可能是由于外源性的IL-27与
IL-27受体的中和作用不能激活细胞相关信号。

Abstract: Objective To investigate the effects of interleukin-27 (IL-27) and its receptor (WSX-1) on the proliferation,
transformation and collagen synthesis of the mouse lung fibroblasts. Methods Cultured mouse lung fibroblasts were treated
with TGF-β1, recombinant murine IL-27, a IL-27 receptor (IL-27R) overexpression vector IL-27R/pCDNA3.1, IL-27 and IL-27R,
or all the 3 combined. MTT assay was used to assess the proliferation of the cells, and RT-PCR and Western blotting were
employed to examine the mRNA and protein expressions of a-smooth muscle actin (α-SMA) and types I and III collagen;
immunofluorescence assay was used to test the expression and location of α-SMA. Results TGF-β1 promoted the cell
proliferation and obviously enhanced α-SMA expression and types I and III collagen synthesis in the fibroblasts. Both IL-27
and IL-27R significantly inhibited the proliferation of the pulmonary fibroblasts and obviously decreased their α-SMA
expression and types I and III collagen synthesis, but when combined,they produced no obvious inhibitory effect on
TGF-1-induced proliferation and transformation of pulmonary fibroblasts. Conclusion Both IL-27 and IL-27R alone can
suppress the proliferation, transformation, and collagen synthesis of mouse pulmonary fibroblasts, but their combined
treatment produces no such inhibitory effect because of the neutralization of exogenous IL-27 by IL-27R to result in the failure
of activating the cell signaling pathways.