Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries

Jesus Prieto-Lloret; Vladimir Snetkov; Yasin Mohammad Shaifta; Inmaculada Docio; Michelle Connolly; Charles Edward MacKay; Gregory Alan Knock; Jeremy Patrick Thomas Ward; Philip Irving Aaronson

doi:10.1152/ajplung.00283.2016

Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries

Jesus Prieto-Lloret, Vladimir Snetkov, Yasin Mohammad Shaifta, Inmaculada Docio, Michelle Connolly, Charles Edward MacKay, Gregory Alan Knock, Jeremy Patrick Thomas Ward, Philip Irving Aaronson^*

^*Corresponding author for this work

King's College London

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Abstract

Application of H₂S (“sulfide”) elicits a complex contraction in rat pulmonary arteries (PAs) comprising a small transient contraction (phase 1; Ph1) followed by relaxation and then a second, larger, and more sustained contraction (phase 2; Ph2). We investigated the mechanisms causing this response using isometric myography in rat second-order PAs, with Na₂S as a sulfide donor. Both phases of contraction to 1,000 μM Na₂S were attenuated by the pan-PKC inhibitor Gö6983 (3 μM) and by 50 μM ryanodine; the Ca²⁺ channel blocker nifedipine (1 μM) was without effect. Ph2 was attenuated by the mitochondrial complex III blocker myxothiazol (1 μM), the NADPH oxidase (NOX) blocker VAS2870 (10 μM), and the antioxidant TEMPOL (3 mM) but was unaffected by the complex I blocker rotenone (1 μM). The bath sulfide concentration, measured using an amperometric sensor, decreased rapidly following Na₂S application, and the peak of Ph2 occurred when this had fallen to ~50 μM. Sulfide caused a transient increase in NAD(P)H autofluorescence, the offset of which coincided with development of the Ph2 contraction. Sulfide also caused a brief mitochondrial hyperpolarization (assessed using tetramethylrhodamine ethyl ester), followed immediately by depolarization and then a second more prolonged hyperpolarization, the onset of which was temporally correlated with the Ph2 contraction. Sulfide application to cultured PA smooth muscle cells increased reactive oxygen species (ROS) production (recorded using L012); this was absent when the mitochondrial flavoprotein sulfide-quinone oxoreductase (SQR) was knocked down using small interfering RNA. We propose that the Ph2 contraction is largely caused by SQR-mediated sulfide metabolism, which, by donating electrons to ubiquinone, increases electron production by complex III and thereby ROS production.

Original language	English
Pages (from-to)	L670-L685
Journal	American Journal of Physiology - Lung Cellular and Molecular Physiology
Volume	314
Issue number	4
Early online date	5 Apr 2018
DOIs	https://doi.org/10.1152/ajplung.00283.2016
Publication status	Published - Apr 2018

Keywords

hydrogen sulfide; mitochondria; protein kinase C; pulmonary artery; rat; reactive oxygen species; sulfide-quinone oxoreductase

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Role of reactive oxygen_PRIETO-LLORET_PublishedApril2018_GOLD VoR (CC BY)Final published version, 1.04 MBLicence: CC BY

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Prieto-Lloret, J., Snetkov, V., Shaifta, Y. M., Docio, I., Connolly, M., MacKay, C. E., Knock, G. A., Ward, J. P. T., & Aaronson, P. I. (2018). Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries. American Journal of Physiology - Lung Cellular and Molecular Physiology, 314(4), L670-L685. https://doi.org/10.1152/ajplung.00283.2016

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title = "Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries",

abstract = "Application of H2S (“sulfide”) elicits a complex contraction in rat pulmonary arteries (PAs) comprising a small transient contraction (phase 1; Ph1) followed by relaxation and then a second, larger, and more sustained contraction (phase 2; Ph2). We investigated the mechanisms causing this response using isometric myography in rat second-order PAs, with Na2S as a sulfide donor. Both phases of contraction to 1,000 μM Na2S were attenuated by the pan-PKC inhibitor G{\"o}6983 (3 μM) and by 50 μM ryanodine; the Ca2+ channel blocker nifedipine (1 μM) was without effect. Ph2 was attenuated by the mitochondrial complex III blocker myxothiazol (1 μM), the NADPH oxidase (NOX) blocker VAS2870 (10 μM), and the antioxidant TEMPOL (3 mM) but was unaffected by the complex I blocker rotenone (1 μM). The bath sulfide concentration, measured using an amperometric sensor, decreased rapidly following Na2S application, and the peak of Ph2 occurred when this had fallen to ~50 μM. Sulfide caused a transient increase in NAD(P)H autofluorescence, the offset of which coincided with development of the Ph2 contraction. Sulfide also caused a brief mitochondrial hyperpolarization (assessed using tetramethylrhodamine ethyl ester), followed immediately by depolarization and then a second more prolonged hyperpolarization, the onset of which was temporally correlated with the Ph2 contraction. Sulfide application to cultured PA smooth muscle cells increased reactive oxygen species (ROS) production (recorded using L012); this was absent when the mitochondrial flavoprotein sulfide-quinone oxoreductase (SQR) was knocked down using small interfering RNA. We propose that the Ph2 contraction is largely caused by SQR-mediated sulfide metabolism, which, by donating electrons to ubiquinone, increases electron production by complex III and thereby ROS production.",

keywords = "hydrogen sulfide; mitochondria; protein kinase C; pulmonary artery; rat; reactive oxygen species; sulfide-quinone oxoreductase",

author = "Jesus Prieto-Lloret and Vladimir Snetkov and Shaifta, {Yasin Mohammad} and Inmaculada Docio and Michelle Connolly and MacKay, {Charles Edward} and Knock, {Gregory Alan} and Ward, {Jeremy Patrick Thomas} and Aaronson, {Philip Irving}",

year = "2018",

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doi = "10.1152/ajplung.00283.2016",

language = "English",

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Prieto-Lloret, J , Snetkov, V , Shaifta, YM, Docio, I, Connolly, M , MacKay, CE , Knock, GA , Ward, JPT & Aaronson, PI 2018, 'Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 314, no. 4, pp. L670-L685. https://doi.org/10.1152/ajplung.00283.2016

Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries. / Prieto-Lloret, Jesus ; Snetkov, Vladimir ; Shaifta, Yasin Mohammad et al.
In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 314, No. 4, 04.2018, p. L670-L685.

Research output: Contribution to journal › Article › peer-review

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T1 - Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries

AU - Prieto-Lloret, Jesus

AU - Snetkov, Vladimir

AU - Shaifta, Yasin Mohammad

AU - Docio, Inmaculada

AU - Connolly, Michelle

AU - MacKay, Charles Edward

AU - Knock, Gregory Alan

AU - Ward, Jeremy Patrick Thomas

AU - Aaronson, Philip Irving

PY - 2018/4

Y1 - 2018/4

N2 - Application of H2S (“sulfide”) elicits a complex contraction in rat pulmonary arteries (PAs) comprising a small transient contraction (phase 1; Ph1) followed by relaxation and then a second, larger, and more sustained contraction (phase 2; Ph2). We investigated the mechanisms causing this response using isometric myography in rat second-order PAs, with Na2S as a sulfide donor. Both phases of contraction to 1,000 μM Na2S were attenuated by the pan-PKC inhibitor Gö6983 (3 μM) and by 50 μM ryanodine; the Ca2+ channel blocker nifedipine (1 μM) was without effect. Ph2 was attenuated by the mitochondrial complex III blocker myxothiazol (1 μM), the NADPH oxidase (NOX) blocker VAS2870 (10 μM), and the antioxidant TEMPOL (3 mM) but was unaffected by the complex I blocker rotenone (1 μM). The bath sulfide concentration, measured using an amperometric sensor, decreased rapidly following Na2S application, and the peak of Ph2 occurred when this had fallen to ~50 μM. Sulfide caused a transient increase in NAD(P)H autofluorescence, the offset of which coincided with development of the Ph2 contraction. Sulfide also caused a brief mitochondrial hyperpolarization (assessed using tetramethylrhodamine ethyl ester), followed immediately by depolarization and then a second more prolonged hyperpolarization, the onset of which was temporally correlated with the Ph2 contraction. Sulfide application to cultured PA smooth muscle cells increased reactive oxygen species (ROS) production (recorded using L012); this was absent when the mitochondrial flavoprotein sulfide-quinone oxoreductase (SQR) was knocked down using small interfering RNA. We propose that the Ph2 contraction is largely caused by SQR-mediated sulfide metabolism, which, by donating electrons to ubiquinone, increases electron production by complex III and thereby ROS production.

AB - Application of H2S (“sulfide”) elicits a complex contraction in rat pulmonary arteries (PAs) comprising a small transient contraction (phase 1; Ph1) followed by relaxation and then a second, larger, and more sustained contraction (phase 2; Ph2). We investigated the mechanisms causing this response using isometric myography in rat second-order PAs, with Na2S as a sulfide donor. Both phases of contraction to 1,000 μM Na2S were attenuated by the pan-PKC inhibitor Gö6983 (3 μM) and by 50 μM ryanodine; the Ca2+ channel blocker nifedipine (1 μM) was without effect. Ph2 was attenuated by the mitochondrial complex III blocker myxothiazol (1 μM), the NADPH oxidase (NOX) blocker VAS2870 (10 μM), and the antioxidant TEMPOL (3 mM) but was unaffected by the complex I blocker rotenone (1 μM). The bath sulfide concentration, measured using an amperometric sensor, decreased rapidly following Na2S application, and the peak of Ph2 occurred when this had fallen to ~50 μM. Sulfide caused a transient increase in NAD(P)H autofluorescence, the offset of which coincided with development of the Ph2 contraction. Sulfide also caused a brief mitochondrial hyperpolarization (assessed using tetramethylrhodamine ethyl ester), followed immediately by depolarization and then a second more prolonged hyperpolarization, the onset of which was temporally correlated with the Ph2 contraction. Sulfide application to cultured PA smooth muscle cells increased reactive oxygen species (ROS) production (recorded using L012); this was absent when the mitochondrial flavoprotein sulfide-quinone oxoreductase (SQR) was knocked down using small interfering RNA. We propose that the Ph2 contraction is largely caused by SQR-mediated sulfide metabolism, which, by donating electrons to ubiquinone, increases electron production by complex III and thereby ROS production.

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Role of reactive oxygen species and sulfide-quinone oxoreductase in hydrogen sulfide-induced contraction of rat pulmonary arteries

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