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Fast relaxation and desensitization of angiotensin II contraction in the pulmonary artery via AT1R and Akt-mediated phosphorylation of muscular eNOS

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Abstract

Angiotensin II (AngII) triggers a transient contraction of pulmonary arteries (PAs) followed by protracted desensitization. Based on the unconventional eNOS expression in PA smooth muscle cells (PASMCs), we hypothesized that activation of smooth muscle eNOS by AngII might be responsible for fast relaxation and tachyphylaxis. Using dual-wire myograph, mechanically endothelium-denuded rat PA [E(−)PA] showed AngII concentration–dependent transient contractions (ΔTAngII, 95% decay within 1 min), which were abolished by losartan (AT1R antagonist). Neither PD123319 (AT2R antagonist) nor A779 (MasR antagonist) affected ΔTAngII. When the vessels were pretreated with L-NAME (NOS inhibitor), ODQ (guanylate cyclase inhibitor), or KT5823 (PKG inhibitor), ΔTAngII of E(−)PA became larger and sustained, whereas nNOS or iNOS inhibitors had no such effect. Immunoblotting of human PASMCs (hPASMCs) also showed eNOS expression, and AngII treatment induced activating phosphorylations of Ser1177 in eNOS and of Ser473 in Akt (Ser/Thr protein kinase B), an upstream signal of eNOS phosphorylation. In addition, L-NAME co-treatment promoted AngII-induced Ser19 phosphorylation of myosin light chain. In hPASMCs, AngII abolished plasma membrane expression of AT1R, and recovery by washout took more than 1 h. Consistent with the data from hPASMCs, the second application of AngII to E(−)PA did not induce contraction, and significant recovery of ΔTAngII required prolonged washout (> 2 h) in the myography study. L-NAME treatment before the second application facilitated recovery of ΔTAngII. Muscular eNOS plays an auto-inhibitory role in ΔTAngII of PAs. The molecular changes investigated in hPASMCs revealed eNOS phosphorylation and internalization of AT1R by AngII. We propose that the rat PA smooth muscle eNOS-induced lusitropy and slow recovery of AT1R from tachyphylaxis might counterbalance the excessive contractile response to AngII, contributing to the distinctive low-pressure pulmonary circulation.

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Acknowledgments

We greatly appreciate the experimental contribution by Mr. Rany Vorn (Chung-Ang University College of Nursing) in the wire myography study.

Funding

This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korea government (NRF-2018R1A5A2025964 and NRF-2018R1D1A1B07048998) to S.J.K.

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  1. Hae Jin Kim and Ji Hyun Jang contributed equally to this work.

Authors and Affiliations

  1. Department of Physiology, Seoul National University College of Medicine, Seoul, 03080, South Korea

    Hae Jin Kim, Ji Hyun Jang, Yin Hua Zhang & Sung Joon Kim

  2. Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea

    Hae Jin Kim, Ji Hyun Jang, Yin Hua Zhang & Sung Joon Kim

  3. Ischemic/Hypoxic Disease Institute, Seoul National University College of Medicine, Seoul, 03080, South Korea

    Yin Hua Zhang & Sung Joon Kim

  4. Chung-Ang University Red Cross College of Nursing, Seoul, 100-031, South Korea

    Hae Young Yoo

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Kim, H.J., Jang, J.H., Zhang, Y.H. et al. Fast relaxation and desensitization of angiotensin II contraction in the pulmonary artery via AT1R and Akt-mediated phosphorylation of muscular eNOS. Pflugers Arch - Eur J Physiol 471, 1317–1330 (2019). https://doi.org/10.1007/s00424-019-02305-z

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