PDE4抑制剂罗氟司特通过激活PKA和Epac双途径，保护心肌细胞免受NO诱导的凋亡。,Cellular Signalling

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PDE4抑制剂罗氟司特通过激活PKA和Epac双途径，保护心肌细胞免受NO诱导的凋亡。
Cellular Signalling ( IF 4.4 ) Pub Date : 2007-12-28 , DOI: 10.1016/j.cellsig.2007.12.011
Hyun-Jeong Kwak ₁ , Kyoung-Mi Park , Hye-Eun Choi , Kyung-Sook Chung , Hyun-Joung Lim , Hyun-Young Park

Affiliation

心肌细胞凋亡在心肌梗塞中起重要作用，而cAMP在调控心肌细胞凋亡中至关重要。磷酸二酯酶4（PDE4）抑制剂可通过抑制PDE4来阻断cAMP的水解，是新型抗炎药的有吸引力的候选药物。但是，其在心血管疾病和心肌细胞凋亡中的功能尚不清楚。因此，我们研究了PDE4抑制剂roflumilast是否对NO诱导的H9c2细胞和新生大鼠心肌细胞（NRCM）的凋亡发挥保护作用，重点研究了cAMP下游分子，例如蛋白激酶A（PKA）和直接激活的交换蛋白由cAMP（Epac）提供。根据我们的数据，在H9c2细胞和NRCM中，罗氟司特治疗可增加细胞内cAMP含量。罗氟司特可抑制SNP诱导的细胞凋亡，PKA特异性抑制剂H-89和KT-5720可逆转这种作用。此外，PKA特异性活化剂N（6）-苯甲酰腺苷3'，5-环一磷酸（N（6）Bz-cAMP）模仿了罗氟司特的作用。罗氟司特的CREB磷酸化也被H-89抑制，表明罗氟司特通过PKA依赖性途径保护SNP诱导的细胞凋亡。罗氟司特增加了Epac1 / GTP-Rap1的活性，Epac1 siRNA转染取消了保护作用，表明Epac信号转导也参与了这种保护反应。作为支持，Epac特异性激活剂8-（4-氯苯硫基）-2'-O-甲基腺苷-3'，5'-环一磷酸酯（8CPT-2Me-cAMP）保护SNP诱导的细胞凋亡。PI3K / Akt抑制剂LY294002阻止罗氟司特诱导的Akt磷酸化和保护作用。此外，用siRNA抑制Epac1对罗氟司特诱导的CREB磷酸化没有影响，而抑制了Akt磷酸化，暗示Akt磷酸化受Epac途径调控。另外，还观察到咯利普兰和西洛司特具有与罗氟司特相似的作用。总之，我们的数据表明罗氟司特同时通过cAMP-PKA / CREB和Epac / Akt依赖性途径保护NO诱导的细胞凋亡。我们的研究表明，PDE4抑制剂罗氟司特有可能作为抗心肌缺血/再灌注（I / R）损伤的潜在治疗剂。暗示Akt磷酸化受Epac途径调控。另外，还观察到咯利普兰和西洛司特具有与罗氟司特相似的作用。总之，我们的数据表明罗氟司特通过cAMP-PKA / CREB和Epac / Akt依赖性途径保护NO诱导的细胞凋亡。我们的研究表明，PDE4抑制剂罗氟司特有可能作为抗心肌缺血/再灌注（I / R）损伤的潜在治疗剂。暗示Akt磷酸化受Epac途径调控。另外，还观察到咯利普兰和西洛司特具有与罗氟司特相似的作用。总之，我们的数据表明罗氟司特通过cAMP-PKA / CREB和Epac / Akt依赖性途径保护NO诱导的细胞凋亡。我们的研究表明，PDE4抑制剂罗氟司特有可能作为抗心肌缺血/再灌注（I / R）损伤的潜在治疗剂。

"点击查看英文标题和摘要"

PDE4 inhibitor, roflumilast protects cardiomyocytes against NO-induced apoptosis via activation of PKA and Epac dual pathways.

Myocyte apoptosis plays an important role in myocardial infarction and cAMP is crucial in the regulation of myocyte apoptosis. Phosphodiesterase-4 (PDE4) inhibitor blocks the hydrolysis of cAMP via inhibition of PDE4 and is attractive candidate for novel anti-inflammatory drugs. However, its function in cardiovascular diseases and cardiomyocyte apoptosis is unclear. Therefore, we investigated whether roflumilast, a PDE4 inhibitor, exerts protective effect against NO-induced apoptosis in both of H9c2 cells and neonatal rat cardiomyocytes (NRCMs), focusing on cAMP downstream molecules such as protein kinase A (PKA) and exchange protein directly activated by cAMP (Epac). According to our data, intracellular cAMP was increased by roflumilast treatment in H9c2 cells and NRCMs. Roflumilast inhibited SNP-induced apoptosis and this effect was reversed by PKA specific inhibitor H-89 and KT-5720. In addition, PKA specific activator N(6)-benzoyladenosine 3',5-cyclic monophosphate (N(6)Bz-cAMP) mimicked the effects of roflumilast. CREB phosphorylation by roflumilast was also inhibited by H-89, indicating that roflumilast protects SNP-induced apoptosis via PKA-dependent pathway. Roflumilast increased Epac1/GTP-Rap1 and the protective effect was abolished by Epac1 siRNA transfection, demonstrating that Epac signaling was also involved in this protective response. In support, Epac specific activator 8-(4-chlrorophenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate (8CPT-2Me-cAMP) protected SNP-induced apoptosis. PI3K/Akt inhibitor LY294002 blocked roflumilast-induced Akt phosphorylation and protective effect. Furthermore, inhibition of Epac1 with siRNA had no effect on roflumilast-induced CREB phosphorylation, whereas inhibited Akt phosphorylation, implicating that Akt phosphorylation was regulated by Epac pathway. In addition, it was also observed that rolipram and cilomilast exert similar effects as roflumilast. In summary, our data indicate that roflumilast protects NO-induced apoptosis via both cAMP-PKA/CREB and Epac/Akt-dependent pathway. Our study suggests a possibility of PDE4 inhibitor roflumilast as a potential therapeutic agent against myocardial ischemia/reperfusion (I/R) injury.

更新日期：2019-11-01

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