PMID- 29921306 OWN - NLM STAT- MEDLINE DCOM- 20181029 LR - 20211204 IS - 1465-993X (Electronic) IS - 1465-9921 (Print) IS - 1465-9921 (Linking) VI - 19 IP - 1 DP - 2018 Jun 19 TI - PDGF-BB regulates the pulmonary vascular tone: impact of prostaglandins, calcium, MAPK- and PI3K/AKT/mTOR signalling and actin polymerisation in pulmonary veins of guinea pigs. PG - 120 LID - 10.1186/s12931-018-0829-5 [doi] LID - 120 AB - BACKGROUND: Platelet-derived growth factor (PDGF)-BB and its receptor PDGFR are highly expressed in pulmonary hypertension (PH) and mediate proliferation. Recently, we showed that PDGF-BB contracts pulmonary veins (PVs) and that this contraction is prevented by inhibition of PDGFR-beta (imatinib/SU6668). Here, we studied PDGF-BB-induced contraction and downstream-signalling in isolated perfused lungs (IPL) and precision-cut lung slices (PCLS) of guinea pigs (GPs). METHODS: In IPLs, PDGF-BB was perfused after or without pre-treatment with imatinib (perfused/nebulised), the effects on the pulmonary arterial pressure (P(PA)), the left atrial pressure (P(LA)) and the capillary pressure (P(cap)) were studied and the precapillary (R(pre)) and postcapillary resistance (R(post)) were calculated. Perfusate samples were analysed (ELISA) to detect the PDGF-BB-induced release of prostaglandin metabolites (TXA(2)/PGI(2)). In PCLS, the contractile effect of PDGF-BB was evaluated in pulmonary arteries (PAs) and PVs. In PVs, PDGF-BB-induced contraction was studied after inhibition of PDGFR-alpha/beta, L-Type Ca(2+)-channels, ROCK/PKC, prostaglandin receptors, MAP2K, p38-MAPK, PI3K-alpha/gamma, AKT/PKB, actin polymerisation, adenyl cyclase and NO. Changes of the vascular tone were measured by videomicroscopy. In PVs, intracellular cAMP was measured by ELISA. RESULTS: In IPLs, PDGF-BB increased P(PA), P(cap) and R(post). In contrast, PDGF-BB had no effect if lungs were pre-treated with imatinib (perfused/nebulised). In PCLS, PDGF-BB significantly contracted PVs/PAs which was blocked by the PDGFR-beta antagonist SU6668. In PVs, inhibition of actin polymerisation and inhibition of L-Type Ca(2+)-channels reduced PDGF-BB-induced contraction, whereas inhibition of ROCK/PKC had no effect. Blocking of EP(1/3)- and TP-receptors or inhibition of MAP2K-, p38-MAPK-, PI3K-alpha/gamma- and AKT/PKB-signalling prevented PDGF-BB-induced contraction, whereas inhibition of EP(4) only slightly reduced it. Accordingly, PDGF-BB increased TXA(2) in the perfusate, whereas PGI(2) was increased in all groups after 120 min and inhibition of IP-receptors did not enhance PDGF-BB-induced contraction. Moreover, PDGF-BB increased cAMP in PVs and inhibition of adenyl cyclase enhanced PDGF-BB-induced contraction, whereas inhibition of NO-formation only slightly increased it. CONCLUSIONS: PDGF-BB/PDGFR regulates the pulmonary vascular tone by the generation of prostaglandins, the increase of calcium, the activation of MAPK- or PI3K/AKT/mTOR signalling and actin remodelling. More insights in PDGF-BB downstream-signalling may contribute to develop new therapeutics for PH. FAU - Rieg, Annette D AU - Rieg AD AUID- ORCID: 0000-0002-7043-5494 AD - Department of Anaesthesiology, Medical Faculty RWTH-Aachen, Aachen, Germany. arieg@ukaachen.de. FAU - Suleiman, Said AU - Suleiman S AD - Institute of Pharmacology and Toxicology, Medical Faculty RWTH-Aachen, Aachen, Germany. FAU - Anker, Carolin AU - Anker C AD - Institute of Pharmacology and Toxicology, Medical Faculty RWTH-Aachen, Aachen, Germany. FAU - Verjans, Eva AU - Verjans E AD - Institute of Pharmacology and Toxicology, Medical Faculty RWTH-Aachen, Aachen, Germany. FAU - Rossaint, Rolf AU - Rossaint R AD - Department of Anaesthesiology, Medical Faculty RWTH-Aachen, Aachen, Germany. FAU - Uhlig, Stefan AU - Uhlig S AD - Institute of Pharmacology and Toxicology, Medical Faculty RWTH-Aachen, Aachen, Germany. FAU - Martin, Christian AU - Martin C AD - Institute of Pharmacology and Toxicology, Medical Faculty RWTH-Aachen, Aachen, Germany. LA - eng GR - 109/14 (691440)/RWTH Aachen University (DE)/International PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20180619 PL - England TA - Respir Res JT - Respiratory research JID - 101090633 RN - 0 (Actins) RN - 0 (Angiogenesis Inducing Agents) RN - 0 (Phosphoinositide-3 Kinase Inhibitors) RN - 0 (Prostaglandins) RN - 0 (Protein Kinase Inhibitors) RN - 0 (Proto-Oncogene Proteins c-sis) RN - 1B56C968OA (Becaplermin) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - SY7Q814VUP (Calcium) SB - IM MH - Actins/*metabolism MH - Angiogenesis Inducing Agents/pharmacology MH - Animals MH - Becaplermin MH - Calcium/metabolism MH - Female MH - Guinea Pigs MH - MAP Kinase Signaling System/*physiology MH - Phosphatidylinositol 3-Kinases/metabolism MH - Phosphoinositide-3 Kinase Inhibitors MH - Polymerization/drug effects MH - Prostaglandins/metabolism MH - Protein Kinase Inhibitors/pharmacology MH - Proto-Oncogene Proteins c-akt/metabolism MH - Proto-Oncogene Proteins c-sis/*pharmacology MH - Pulmonary Veins/drug effects/*physiology MH - TOR Serine-Threonine Kinases/metabolism MH - Vasomotor System/drug effects/*metabolism PMC - PMC6009037 COIS- COMPETING INTEREST: The authors declare that they have no competing interests. ETHICS APPROVAL AND CONSENT TO PARTICIPATE: Female Dunkin Hartley GPs (350 +/- 50 g) were delivered from Charles River (Sulzfeld, Germany). All animal experiments were approved by the Landesamt fur Natur, Umwelt und Verbraucherschutz Nordrhein-Westfalen (ID: 84-02.04.2013A146, 8.87-51.05.20.10.245 and 50066A4) and strictly performed according to the rules of the Directive 2010/63/EU of the European Parliament. Intraperitoneal anaesthesia was performed (pentobarbital: 95 mg kg(- 1), Narcoren: Garbsen, Germany) and verified by missing reflexes, afterwards the animal was exsanguinated, CONSENT FOR PUBLICATION: Not applicable PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2018/06/21 06:00 MHDA- 2018/10/30 06:00 PMCR- 2018/06/19 CRDT- 2018/06/21 06:00 PHST- 2018/01/10 00:00 [received] PHST- 2018/06/13 00:00 [accepted] PHST- 2018/06/21 06:00 [entrez] PHST- 2018/06/21 06:00 [pubmed] PHST- 2018/10/30 06:00 [medline] PHST- 2018/06/19 00:00 [pmc-release] AID - 10.1186/s12931-018-0829-5 [pii] AID - 829 [pii] AID - 10.1186/s12931-018-0829-5 [doi] PST - epublish SO - Respir Res. 2018 Jun 19;19(1):120. doi: 10.1186/s12931-018-0829-5.