Zhao L, Xiao K, Wang H, Wang Z, Sun L, Zhang F, Zhang X, Tang F, He W
Department of Rheumatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Clin Exp Immunol. 2009 Aug;157(2):310-5. doi: 10.1111/j.1365-2249.2009.03962.x.
The objective of this study was to investigate the effects of thalidomide (THD) on interstitial lung fibrosis (ILF). In vitro, human fetal lung fibroblast (HFL-F) to myofibroblast (MF) trans-differentiation was induced by transforming growth factor (TGF)-beta1. The effects of THD on trans-differentiation process or differentiated MF were evaluated by measuring hydroxyproline (HYP) content by alkaline hydrolysis colorimetry, alpha-smooth muscle actin (alpha-SMA) protein by Western blot and alpha-SMA and pro-collagen III mRNA expressions by semi-quantitative reverse transcription-polymerase chain reaction; in vivo, a mouse model of ILF was generated by daily subcutaneous injection of bleomycin (BLM) in female C3H mice. Gastric perfusion of THD began 1 week prior to injection and lasted for 8 weeks. Lung specimens were harvested at different time-points (1, 4, 6 and 8 weeks) for pathology and immunohistochemistry examination. The HYP content, alpha-SMA and pro-collagen III mRNA expressions were also assessed. THD inhibited the up-regulation of HYP protein, pro-collagen III mRNA and alpha-SMA protein induced by TGF-beta1 in HFL-F cells, and additionally inhibited pro-collagen III mRNA expression on trans-differentiated MF. THD reduced HYP synthesis in the lung tissues of BLM-treated mice at week 4, and slightly reduced the numbers of alpha-SMA-positive cells. THD had an effect on ILF models both in vitro and in vivo.
本研究的目的是探讨沙利度胺（THD）对间质性肺纤维化（ILF）的影响。在体外，通过转化生长因子（TGF）-β1诱导人胎儿肺成纤维细胞（HFL-F）向肌成纤维细胞（MF）转分化。通过碱性水解比色法测量羟脯氨酸（HYP）含量、蛋白质免疫印迹法检测α-平滑肌肌动蛋白（α-SMA）蛋白以及半定量逆转录-聚合酶链反应检测α-SMA和Ⅲ型前胶原mRNA表达，来评估THD对转分化过程或分化后的MF的影响；在体内，通过每天给雌性C3H小鼠皮下注射博来霉素（BLM）建立ILF小鼠模型。在注射前1周开始胃灌注THD，持续8周。在不同时间点（1、4、6和8周）采集肺组织标本进行病理学和免疫组织化学检查。同时评估HYP含量、α-SMA和Ⅲ型前胶原mRNA表达。THD抑制TGF-β1诱导的HFL-F细胞中HYP蛋白、Ⅲ型前胶原mRNA和α-SMA蛋白的上调，并且还抑制转分化后的MF中Ⅲ型前胶原mRNA的表达。THD在第4周时降低了BLM处理小鼠肺组织中HYP的合成，并略微减少了α-SMA阳性细胞的数量。THD在体外和体内对ILF模型均有作用。
