Crosstalk between diacylglycerol kinase and protein kinase A in the regulation of airway smooth muscle cell proliferation.
Crosstalk between diacylglycerol kinase and protein kinase A in the regulation of airway smooth muscle cell proliferation.
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DOI：
10.1186/s12931-023-02465-8
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发表时间：
2023-06-10
影响因子：
5.8
通讯作者：
中科院分区：
文献类型：
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二酰基甘油激酶（DGK）通过将二酰基甘油（DAG）转化为磷脂酸来调节细胞内信号传导和功能。我们以前证明，DGK抑制减弱气道平滑肌（ASM）细胞增殖，然而，介导这种效果的机制还没有很好地建立。鉴于蛋白激酶A（PKA）的能力，以影响抑制ASM细胞的生长，有丝分裂原，我们采用多种分子和药理学方法来检查PKA的抑制丝分裂原诱导的ASM细胞增殖的小分子DGK抑制剂I（DGK I）的推定作用。我们使用CyQUANT™ NF测定法测定细胞增殖，使用免疫印迹法测定蛋白质表达和磷酸化，并通过ELISA测定前列腺素E2（PGE 2）分泌。用血小板衍生生长因子（PDGF）或PDGF + DGK I刺激稳定表达GFP或PKI-GFP（PKA抑制肽-GFP嵌合体）的ASM细胞，并评估细胞增殖。DGK抑制可减少表达GFP的细胞中的ASM细胞增殖，但在表达PKI-GFP的细胞中则不然。随着时间的推移，DGK抑制增加环氧合酶II（COXII）表达和PGE 2分泌，以促进PKA活化，如通过（PKA底物）VASP和CREB的磷酸化增加所证明的。在用pan-PKC（Bis I）、MEK（U 0126）或ERK 2（ERK 11 e）抑制剂预处理的细胞中，COXII表达和PKA活化显著降低，表明PKC和ERK在通过DGK抑制的COXII-PGE 2介导的PKA信号传导活化中的作用。我们的研究提供了对DGK在ASM细胞中调节的分子途径（DAG-PKC/ERK-COXII-PGE 2-PKA）的深入了解，并将DGK确定为减轻ASM细胞增殖的潜在治疗靶点，该细胞增殖有助于哮喘中的气道重塑。
Diacylglycerol kinase (DGK) regulates intracellular signaling and functions by converting diacylglycerol (DAG) into phosphatidic acid. We previously demonstrated that DGK inhibition attenuates airway smooth muscle (ASM) cell proliferation, however, the mechanisms mediating this effect are not well established. Given the capacity of protein kinase A (PKA) to effect inhibition of ASM cells growth in response to mitogens, we employed multiple molecular and pharmacological approaches to examine the putative role of PKA in the inhibition of mitogen-induced ASM cell proliferation by the small molecular DGK inhibitor I (DGK I). We assayed cell proliferation using CyQUANT™ NF assay, protein expression and phosphorylation using immunoblotting, and prostaglandin E2 (PGE2) secretion by ELISA. ASM cells stably expressing GFP or PKI-GFP (PKA inhibitory peptide-GFP chimera) were stimulated with platelet-derived growth factor (PDGF), or PDGF + DGK I, and cell proliferation was assessed. DGK inhibition reduced ASM cell proliferation in cells expressing GFP, but not in cells expressing PKI-GFP. DGK inhibition increased cyclooxygenase II (COXII) expression and PGE2 secretion over time to promote PKA activation as demonstrated by increased phosphorylation of (PKA substrates) VASP and CREB. COXII expression and PKA activation were significantly decreased in cells pre-treated with pan-PKC (Bis I), MEK (U0126), or ERK2 (Vx11e) inhibitors suggesting a role for PKC and ERK in the COXII-PGE2-mediated activation of PKA signaling by DGK inhibition. Our study provides insight into the molecular pathway (DAG-PKC/ERK-COXII-PGE2-PKA) regulated by DGK in ASM cells and identifies DGK as a potential therapeutic target for mitigating ASM cell proliferation that contributes to airway remodeling in asthma.
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影响因子：
16.6
作者：
Katti, Sachin S.;Krieger, Inna, V;Ann, Jihyae;Lee, Jeewoo;Sacchettini, James C.;Igumenova, Tatyana I.
通讯作者：
Igumenova, Tatyana I.
气道平滑肌上的苦味受体通过局部钙信号传导和反向阻塞来支气管扩张。
影响因子：
82.9
作者：
通讯作者：
--
DOI：
10.1152/ajplung.00013.2006
发表时间：
2006-09-01
影响因子：
4.9
作者：
Gosens, Reinoud;Stelmack, Gerald L.;Halayko, Andrew J.
通讯作者：
Halayko, Andrew J.
影响因子：
4.8
作者：
Defnet, Amy E.;Huang, Weiliang;Deshpande, Deepak A.
通讯作者：
Deshpande, Deepak A.
DOI：
10.1152/ajplung.1999.277.5.l943
发表时间：
1999-11-01
影响因子：
4.9
作者：
Laporte, JD;Moore, PE;Shore, SA
通讯作者：
Shore, SA