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Neuronal nitric oxide synthase modulation of intracellular Ca2+ handling overrides fatty acid potentiation of cardiac inotropy in hypertensive rats

  • Muscle physiology
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Abstract

Cardiac neuronal nitric oxide synthase (nNOS) is an important molecule that regulates intracellular Ca2+ homeostasis and contractility of healthy and diseased hearts. Here, we examined the effects of nNOS on fatty acid (FA) regulation of left ventricular (LV) myocyte contraction in sham and angiotensin II (Ang II)-induced hypertensive (HTN) rats. Our results showed that palmitic acid (PA, 100 μM) increased the amplitudes of sarcomere shortening and intracellular ATP in sham but not in HTN despite oxygen consumption rate (OCR) was increased by PA in both groups. Carnitine palmitoyltransferase-1 inhibitor, etomoxir (ETO), reduced OCR and ATP with PA in sham and HTN but prevented PA potentiation of sarcomere shortening only in sham. PA increased nNOS-derived NO only in HTN. Inhibition of nNOS with S-methyl-l-thiocitrulline (SMTC) prevented PA-induced OCR and restored PA potentiation of myocyte contraction in HTN. Mechanistically, PA increased intracellular Ca2+ transient ([Ca2+]i) without changing Ca2+ influx via L-type Ca2+ channel (I-LTCC) and reduced myofilament Ca2+ sensitivity in sham. nNOS inhibition increased [Ca2+]i, I-LTCC and reduced myofilament Ca2+ sensitivity prior to PA supplementation; as such, normalized PA increment of [Ca2+]i. In HTN, PA reduced I-LTCC without affecting [Ca2+]i or myofilament Ca2+ sensitivity. However, PA increased I-LTCC, [Ca2+]i and reduced myofilament Ca2+ sensitivity following nNOS inhibition. Myocardial FA oxidation (18F-fluoro-6-thia-heptadecanoic acid, 18F-FTHA) was comparable between groups, but nNOS inhibition increased it only in HTN. Collectively, PA increases myocyte contraction through stimulating [Ca2+]i and mitochondrial activity in healthy hearts. PA-dependent cardiac inotropy was limited by nNOS in HTN, predominantly due to its modulatory effect on [Ca2+]i handling.

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Authors and Affiliations

  1. Department of Physiology & Biomedical Sciences, Ischemic/Hypoxic Disease Institute, College of Medicine, Seoul National University, Jongno Gu, 103 Dae Hak Ro, Seoul, 110-799, South Korea

    Chun Li Jin, Ming Zhe Yin, Zai Hao Zhao, Yue Wang, Sung Joon Kim & Yin Hua Zhang

  2. Yan Bian University Hospital, Yanji, Ji Lin, China

    Chun Li Jin, Chun Zi Jin & Yin Hua Zhang

  3. Department of Nuclear Medicine, Seoul National University Hospital, Seoul, South Korea

    Jin Chul Paeng & Keon Wook Kang

  4. Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, College of Medicine or College of Pharmacy, Seoul National University, Seoul, South Korea

    Seunggyun Ha

  5. Department of Physiology, Ulsan College of Medicine, Seoul, South Korea

    Jeong Hoon Lee & Chae Hun Leem

  6. Department of Pharmacology & Biomedical Sciences, College of Medicine, Seoul National University, Seoul, South Korea

    Peng Jin & Jong-Wan Park

  7. Division of Cardiovascular Sciences, School of Medical Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK

    Yin Hua Zhang

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  1. Chun Li Jin
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Correspondence to Yin Hua Zhang.

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Ethical approval

The study protocol was in accordance with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996) and also conforms to the Institutional Animal Care and Use Committee (IACUC) in Seoul National University (IACUC approval no.: SNU-101213-1; SNU-160119-4-1).

Conflicts of interests

None.

Sources of funding

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2013068067), the Brain Korea 21 Graduate Programme of the Korean Ministry of Education, Science and Technology, Seoul National University Hospital, the Korean Society of Hypertension (2013), SK Telecom Research Fund (no. 3420130290), and the National Natural Science Foundation of China (NSFC, 31460265).

Electronic supplementary material

ESM 1

Supplementary Fig. 1. Effects of ROS scavenger, tiron, on PA regulation of myocyte contraction in sham and in HTN rats. a–c Representative raw traces and mean values of sarcomere shortening by PA with tiron (1 mM). Sarcomere shortening was remained increased by PA in sham rats and was unaffected by PA in HTN rats, despite tiron pre-treatment. Supplementary Fig. 2. Examination of FA uptake with PET/CT in sham and HTN rats. a. PET/CT images were obtained by using 18F-FTHA in the LV at basal, in the presence of nNOS inhibitor, SMTC (1 mg/kg) and l-NAME (5 mg/kg). b. Averaged mean values of 18F-FTHA uptake in the myocardium/liver between sham and HTN with and without NOS inhibition. nNOS inhibition induced small but significant increase in 18F-FTHA uptake in HTN rats only. l-NAME significantly increased 18F-FTHA uptake in both groups, with an effect significantly higher in sham rats compared to that in HTN rats. Supplementary Fig. 3. PA regulation of the peak ICa-L density and inactivation kinetics with and without nNOS inhibition in sham and HTN rats. LTCC was stimulated by a step depolarization protocol from −60 to +40 mV (10 mV interval) for 200 ms (holding potential, −40 mV). a–d Representative ICa-L density with and without PA before and after nNOS inhibition (SMTC, 100 nM, 30 min–1 h). PA slightly but significantly reduced peak ICa-L density at 0 mV in both groups. nNOS inhibition increased ICa-L density at 0 mV and abolished PA regulation in both groups. e–h Inactivation of ICa-L was slowed by PA in both groups; nNOS inhibition (SMTC, 100 nM, 30 min–1 h) significantly reduced inactivation and abolished PA-dependent responses in sham and in HTN rats. Supplementary Fig. 4. PA regulation of the Ca2+ influx through LTCC. LTCC was stimulated by a step depolarization protocol from −60 to +40 mV (10 mV interval) for 200 ms (holding potential, −40 mV). a & c. Representative ICa-L integral at 0 mV in sham and HTN rats before and after nNOS inhibition with and without PA. b & d. Averaged mean values showed that PA did not change ICa-L integral in sham rats but significantly reduced it in HTN rats. nNOS inhibition increased ICa-L integral with and without PA in both groups and abolished the response of PA in two groups. (PPTX 2.74 MB)

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Jin, C.L., Yin, M.Z., Paeng, J.C. et al. Neuronal nitric oxide synthase modulation of intracellular Ca2+ handling overrides fatty acid potentiation of cardiac inotropy in hypertensive rats. Pflugers Arch - Eur J Physiol 469, 1359–1371 (2017). https://doi.org/10.1007/s00424-017-1991-1

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