Phosphodiesterase type 4 expression and anti-proliferative effects in human pulmonary artery smooth muscle cells.
Phosphodiesterase type 4 expression and anti-proliferative effects in human pulmonary artery smooth muscle cells.
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DOI：
10.1186/1465-9921-7-9
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发表时间：
2006-01-19
影响因子：
5.8
通讯作者：
Wharton J
中科院分区：
文献类型：
--
作者：
Growcott EJ;Spink KG;Ren X;Afzal S;Banner KH;Wharton J
肺动脉高压是一种以远端肺动脉平滑肌细胞增殖和凋亡调节异常为特征的增殖性血管疾病。前列环素（PGI 2）类似物对远端人肺动脉平滑肌细胞（PASMCs）具有抗增殖作用，这依赖于细胞内cAMP刺激。因此，我们试图调查的主要cAMP特异性酶，磷酸二酯酶4型（PDE 4），负责cAMP水解的参与。远端人PASMCs通过外植体培养从肺动脉衍生（n = 14，第3-12代）。通过测量cAMP磷酸二酯酶活性、细胞内cAMP水平、DNA合成、细胞凋亡（通过DNA片段化和核凝聚测量）和基质金属蛋白酶-2和-9（MMP-2、MMP-9）产生来确定对血小板衍生生长因子-BB（5-10 ng/ml）、血清、PGI 2类似物（cicaprost、iloprost）和PDE 4抑制剂（罗氟司特、咯利普兰、西洛司特）的反应。在PASMC分离株中检测到所有四种PDE 4A-D基因的表达。与PDE 3（21.5 ± 2.5%）、PDE 2（15.8 ± 3.4%）或PDE 1活性（14.5 ± 4.2%）相比，PDE 4占PASMC中cAMP特异性水解活性的主要比例（35.9 ± 2.3%，n = 5）。细胞内cAMP水平通过PGI 2类似物增加，并且在用罗氟司特、咯利普兰和西洛司特共处理的细胞中进一步升高。1 μM罗氟司特（49 ± 6%抑制）、咯利普兰（37 ± 6%）和西洛司特（30 ± 4%）可减弱DNA合成，在5 nM环前列素存在下，这些化合物的EC 50值分别为4.4（2.6-6.1）nM（平均值和95%置信区间）、59（36-83）nM和97（66-130）nM。罗氟司特减弱细胞增殖和明胶酶（MMP-2和MMP-9）的产生，并促进PGI 2类似物的抗增殖作用。cAMP激活剂伊洛前列素和毛喉素也诱导细胞凋亡，而罗氟司特没有显着的效果。PDE 4酶在远端人PASMC中表达，cAMP刺激剂对DNA合成、增殖和MMP产生的作用至少部分依赖于PDE 4活性。PDE 4抑制可以提供对人PASMC中cAMP介导的抗增殖作用的更好控制，因此可以证明可用作肺动脉高压的额外治疗。
Pulmonary arterial hypertension is a proliferative vascular disease, characterized by aberrant regulation of smooth muscle cell proliferation and apoptosis in distal pulmonary arteries. Prostacyclin (PGI2) analogues have anti-proliferative effects on distal human pulmonary artery smooth muscle cells (PASMCs), which are dependent on intracellular cAMP stimulation. We therefore sought to investigate the involvement of the main cAMP-specific enzymes, phosphodiesterase type 4 (PDE4), responsible for cAMP hydrolysis. Distal human PASMCs were derived from pulmonary arteries by explant culture (n = 14, passage 3–12). Responses to platelet-derived growth factor-BB (5–10 ng/ml), serum, PGI2 analogues (cicaprost, iloprost) and PDE4 inhibitors (roflumilast, rolipram, cilomilast) were determined by measuring cAMP phosphodiesterase activity, intracellular cAMP levels, DNA synthesis, apoptosis (as measured by DNA fragmentation and nuclear condensation) and matrix metalloproteinase-2 and -9 (MMP-2, MMP-9) production. Expression of all four PDE4A-D genes was detected in PASMC isolates. PDE4 contributed to the main proportion (35.9 ± 2.3%, n = 5) of cAMP-specific hydrolytic activity demonstrated in PASMCs, compared to PDE3 (21.5 ± 2.5%), PDE2 (15.8 ± 3.4%) or PDE1 activity (14.5 ± 4.2%). Intracellular cAMP levels were increased by PGI2 analogues and further elevated in cells co-treated with roflumilast, rolipram and cilomilast. DNA synthesis was attenuated by 1 μM roflumilast (49 ± 6% inhibition), rolipram (37 ± 6%) and cilomilast (30 ± 4%) and, in the presence of 5 nM cicaprost, these compounds exhibited EC50 values of 4.4 (2.6–6.1) nM (Mean and 95% confidence interval), 59 (36–83) nM and 97 (66–130) nM respectively. Roflumilast attenuated cell proliferation and gelatinase (MMP-2 and MMP-9) production and promoted the anti-proliferative effects of PGI2 analogues. The cAMP activators iloprost and forskolin also induced apoptosis, whereas roflumilast had no significant effect. PDE4 enzymes are expressed in distal human PASMCs and the effects of cAMP-stimulating agents on DNA synthesis, proliferation and MMP production is dependent, at least in part, on PDE4 activity. PDE4 inhibition may provide greater control of cAMP-mediated anti-proliferative effects in human PASMCs and therefore could prove useful as an additional therapy for pulmonary arterial hypertension.
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DOI：
10.1165/rcmb.2002-0254oc
发表时间：
2003-07-01
影响因子：
6.4
作者：
Goncharova, EA;Billington, CK;Panettieri, RA
通讯作者：
Panettieri, RA
影响因子：
3.2
作者：
Jones, NA;Boswell-Smith, V;Page, CP
通讯作者：
Page, CP
影响因子：
24
作者：
Humbert, M;Morrell, NW;Rabinovitch, M
通讯作者：
Rabinovitch, M
DOI：
10.1165/ajrcmb.26.2.4695
发表时间：
2002-02-01
影响因子：
6.4
作者：
Clapp, LH;Finney, P;Tinker, A
通讯作者：
Tinker, A
影响因子：
15.9
作者：
McMurtry, MS;Archer, SL;Michelakis, ED
通讯作者：
Michelakis, ED