背景:特发性肺纤维化(IPF)中细胞外基质(ECM)的广泛沉积是由于肺成纤维细胞的过度活化和增殖所致。然而,确切的机制尚不清楚。
目的:本研究关注CTBP1在肺成纤维细胞功能中的作用,阐述了其调控机制,分析了CTBP1与ZEB1的关系。同时,研究了Toosendanin的抗肺纤维化作用及其分子机制。
方法:体外培养人IPF成纤维细胞系(LL-97A和LL-29)和正常成纤维细胞系(LL-24)。用FCS刺激细胞,PDGF-BB,分别为IGF-1和TGF-β1。BrdU检测到细胞增殖。QRT-PCR检测CTBP1和ZEB1的mRNA表达。Westernblot检测COL1A1、COL3A1、LN、FN,和α-SMA蛋白。建立肺纤维化动物模型,分析CTBP1沉默对小鼠肺纤维化及肺功能的影响。
结果:CTBP1在IPF肺成纤维细胞中上调。沉默CTBP1抑制生长因子驱动的肺成纤维细胞增殖和激活。CTBP1的过表达促进生长因子驱动的肺成纤维细胞的增殖和活化。沉默CTBP1可降低肺纤维化小鼠肺纤维化程度。蛋白质印迹,CO-IP,和BrdU测定证实CTBP1与ZEB1相互作用并促进肺成纤维细胞的活化。Toosendanin可以抑制ZEB1/CTBP1蛋白相互作用,进一步抑制肺纤维化的进展。
结论:CTBP1可通过ZEB1促进肺成纤维细胞的活化和增殖。CTBP1通过ZEB1促进肺成纤维细胞活化,从而增加ECM的过度沉积并加重IPF。Toosendanin可能是肺纤维化的潜在治疗方法。本研究结果为阐明肺纤维化的分子机制和开发新的治疗靶点提供了新的依据。
Extensive deposition of extracellular matrix (ECM) in idiopathic pulmonary fibrosis (IPF) is due to hyperactivation and proliferation of pulmonary fibroblasts. However, the exact mechanism is not clear.
This study focused on the role of CTBP1 in lung fibroblast function, elaborated its regulation mechanism, and analyzed the relationship between
CTBP1 and ZEB1. Meanwhile, the antipulmonary fibrosis effect and its molecular mechanism of Toosendanin were studied.
Human IPF fibroblast cell lines (LL-97A and LL-29) and normal fibroblast cell lines (LL-24) were cultured in vitro. The cells were stimulated with FCS, PDGF-BB, IGF-1, and TGF-β1, respectively. BrdU detected cell proliferation. The mRNA expression of
CTBP1 and ZEB1 was detected by QRT-PCR. Western blotting was used to detect the expression of COL1A1, COL3A1, LN, FN, and α-SMA proteins. An animal model of pulmonary fibrosis was established to analyze the effects of CTBP1 silencing on pulmonary fibrosis and lung function in mice.
CTBP1 was up-regulated in IPF lung fibroblasts. Silencing CTBP1 inhibits growth factor-driven proliferation and activation of lung fibroblasts. Overexpression of CTBP1 promotes growth factor-driven proliferation and activation of lung fibroblasts. Silencing
CTBP1 reduced the degree of pulmonary fibrosis in mice with pulmonary fibrosis. Western blot, CO-IP, and BrdU assays confirmed that CTBP1 interacts with ZEB1 and promotes the activation of lung fibroblasts. Toosendanin can inhibit the ZEB1/CTBP1protein interaction and further inhibit the progression of pulmonary fibrosis.
CTBP1 can promote the activation and proliferation of lung fibroblasts through ZEB1. CTBP1 promotes lung fibroblast activation through ZEB1, thereby increasing excessive deposition of ECM and aggravating IPF. Toosendanin may be a potential treatment for pulmonary fibrosis. The results of this study provide a new basis for clarifying the molecular mechanism of pulmonary fibrosis and developing new therapeutic targets.