He Donghong, Natarajan Viswanathan, Stern Randi, Gorshkova Irina A, Solway Julian, Spannhake Ernst Wm, Zhao Yutong
The Section of Pulmonary and Critical Care, Department of Medicine, Center for Integrative Science Building, 929 East 57th Street, Chicago, IL 60637, USA.
Biochem J. 2008 May 15;412(1):153-62. doi: 10.1042/BJ20071649.
We have demonstrated that LPA (lysophosphatidic acid)-induced IL (interleukin)-8 secretion was partly mediated via transactivation of EGFR [EGF (epidermal growth factor) receptor] in HBEpCs (human bronchial epithelial primary cells). The present study provides evidence that LPA-induced transactivation of EGFR regulates COX (cyclo-oxygenase)-2 expression and PGE(2) [PG (prostaglandin) E(2)] release through the transcriptional factor, C/EBPbeta (CCAAT/enhancer-binding protein beta), in HBEpCs. Treatment with LPA (1 microM) stimulated COX-2 mRNA and protein expression and PGE(2) release via G(alphai)-coupled LPARs (LPA receptors). Pretreatment with inhibitors of NF-kappaB (nuclear factor-kappaB), JNK (Jun N-terminal kinase), or down-regulation of c-Jun or C/EBPbeta with specific siRNA (small interference RNA) attenuated LPA-induced COX-2 expression. Downregulation of EGFR by siRNA or pretreatment with the EGFR tyrosine kinase inhibitor, AG1478, partly attenuated LPA-induced COX-2 expression and phosphorylation of C/EBPbeta; however, neither of these factors had an effect on the NF-kappaB and JNK pathways. Furthermore, LPA-induced EGFR transactivation, phosphorylation of C/EBPbeta and COX-2 expression were attenuated by overexpression of a catalytically inactive mutant of PLD2 [PLD (phospholipase D) 2], PLD2-K758R, or by addition of myristoylated PKCzeta [PKC (protein kinase C) zeta] peptide pseudosubstrate. Overexpression of the PLD2-K758R mutant also attenuated LPA-induced phosphorylation and activation of PKCzeta. These results demonstrate that LPA induces COX-2 expression and PGE(2) production through EGFR transactivation-independent activation of transcriptional factors NF-kappaB and c-Jun, and EGFR transactivation-dependent activation of C/EBPbeta in HBEpCs. Since COX-2 and PGE(2) have been shown to be anti-inflammatory in airway inflammation, the present data suggest a modulating and protective role of LPA in regulating innate immunity and remodelling of the airways.
我们已经证明,溶血磷脂酸(LPA)诱导的白细胞介素-8(IL-8)分泌部分是通过人支气管上皮原代细胞(HBEpCs)中表皮生长因子受体(EGFR)的反式激活介导的。本研究提供了证据,表明LPA诱导的EGFR反式激活通过转录因子CCAAT/增强子结合蛋白β(C/EBPβ)调节HBEpCs中环氧合酶-2(COX-2)的表达和前列腺素E2(PGE2)的释放。用LPA(1微摩尔)处理通过Gαi偶联的溶血磷脂酸受体(LPARs)刺激COX-2 mRNA和蛋白表达以及PGE2释放。用核因子κB(NF-κB)抑制剂、Jun氨基末端激酶(JNK)抑制剂预处理,或用特异性小干扰RNA(siRNA)下调c-Jun或C/EBPβ,可减弱LPA诱导的COX-2表达。用siRNA下调EGFR或用EGFR酪氨酸激酶抑制剂AG1478预处理,可部分减弱LPA诱导的COX-2表达和C/EBPβ的磷酸化;然而,这些因素均对NF-κB和JNK途径无影响。此外,通过过表达磷脂酶D2(PLD2)的催化失活突变体PLD2-K758R,或添加肉豆蔻酰化的蛋白激酶Cζ(PKCζ)肽假底物,可减弱LPA诱导的EGFR反式激活、C/EBPβ的磷酸化和COX-2表达。PLD2-K758R突变体的过表达也减弱了LPA诱导的PKCζ的磷酸化和激活。这些结果表明,LPA通过转录因子NF-κB和c-Jun的EGFR反式激活非依赖性激活以及C/EBPβ的EGFR反式激活依赖性激活,诱导HBEpCs中COX-2表达和PGE2产生。由于COX-2和PGE2在气道炎症中已被证明具有抗炎作用,目前的数据表明LPA在调节固有免疫和气道重塑中具有调节和保护作用。