American Journal of Respiratory Cell and Molecular Biology ( IF 6.4 ) Pub Date : 2020-12-01 , DOI: 10.1165/rcmb.2020-0227oc Asres Berhan 1 , Trudi Harris 1 , Jade Jaffar 2 , Fernando Jativa 1, 3 , Shenna Langenbach 1 , Ingrid Lönnstedt 4 , Monther Alhamdoosh 4 , Milica Ng 4 , Peter Lee 3 , Glen Westall 2 , Nick Wilson 4 , Michael Wilson 4 , Alastair G Stewart 1, 5
Pathological changes in the biomechanical environment are implicated in the progression of idiopathic pulmonary fibrosis (IPF). Stiffened matrix augments fibroblast proliferation and differentiation and activates TGF-β1 (transforming growth factor-β1). Stiffened matrix impairs the synthesis of the antifibrogenic lipid mediator prostaglandin E2 (PGE2) and reduces the expression of the rate-limiting prostanoid biosynthetic enzyme cyclooxygenase-2 (COX-2). We now show that prostaglandin E synthase (PTGES), the final enzyme in the PGE2 biosynthetic pathway, is expressed at lower levels in the lungs of patients with IPF. We also show substantial induction of COX-2, PTGES, prostaglandin E receptor 4 (EP4), and cytosolic phospholipase A2 (cPLA2) expression in human lung fibroblasts cultured in soft collagen hydrogels or in spheroids compared with conventional culture on stiff plastic culture plates. Induction of COX-2, cPLA2, and PTGES expression in spheroid cultures was moderately inhibited by the p38 mitogen-activated protein kinase inhibitor SB203580. The induction of prostanoid biosynthetic enzyme expression was accompanied by an increase in PGE2 levels only in non–IPF-derived fibroblast spheroids. Our study reveals an extensive dysregulation of prostanoid biosynthesis and signaling pathways in IPF-derived fibroblasts, which are only partially abrogated by culture in soft microenvironments.
中文翻译:
细胞微环境的硬度调节类花生酸的产生和信号通路。
生物力学环境中的病理变化与特发性肺纤维化(IPF)的进展有关。增强的基质增强了成纤维细胞的增殖和分化并激活了TGF-β1(转化生长因子-β1)。强化的基质会损害抗纤维化脂质介体前列腺素E 2(PGE 2)的合成,并降低限速前列腺素类生物合成酶环氧合酶2(COX-2)的表达。我们现在显示前列腺素E合酶(PTGES),PGE 2生物合成途径中的最终酶,在IPF患者的肺中表达水平较低。我们还显示大量诱导COX-2,PTGES,前列腺素E受体4(EP4)和胞质磷脂酶A 2(cPLA 2)在硬质胶原培养板上与常规培养相比,在软胶原水凝胶或球体中培养的人肺成纤维细胞中的表达。p38丝裂原活化的蛋白激酶抑制剂SB203580适度抑制了球状培养物中COX-2,cPLA 2和PTGES表达的诱导。前列腺素生物合成酶表达的诱导仅在非IPF来源的成纤维细胞球体中PGE 2水平增加。我们的研究揭示了IPF来源的成纤维细胞中前列腺素生物合成和信号通路的广泛失调,而在软微环境中培养只能部分消除这种失调。