Measurement of Reactive Oxygen Species in Cardiovascular Studies

Dikalov, Sergey; Griendling, Kathy K.; Harrison, David G.

doi:10.1161/01.hyp.0000258594.87211.6b

Cited by 465 publications

(433 citation statements)

References 63 publications

(60 reference statements)

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“…It is obvious that the site of accumulation of bright-red fluorescence from oxidized DHE and DNA complex is not a suitable indicator of the location where ROS were produced [37]. In contrast, CM-H 2 DCFDA is able to localize cellular production of ROS within the tissue, and the sensitivity to detect ROS within a single cell, these strengths has made CM-H 2 DCFDA widely used in the measurement of intracellular localized ROS by imaging assay [38,39]. With the help of an Image-Pro Plus analysis software (version 6.0), the intensity of spots with different colors or spectra could be quantified in overlaid fluorescent confocal images.…”

Section: Discussionmentioning

confidence: 99%

Local production of O2− by NAD(P)H oxidase in the sarcoplasmic reticulum of coronary arterial myocytes: cADPR-mediated Ca2+ regulation

Zhang

Jin

Xia

et al. 2008

Cellular Signalling

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The present study was designed to determine whether the sarcoplasmic reticulum (SR) could locally produce superoxide (O 2 .− ) via NAD(P)H oxidase (NOX) in coronary arterial myocytes (CAMs) and to address whether cADPR-RyR/Ca 2+ signaling pathway regulates this local O 2 .− production from the SR. Using confocal microscopic imaging analysis in intact single CAMs, a cell-permeable indicator CM-H 2 DCFDA for dynamic changes in intracellular ROS (in green color) and a highly selective ER-Tracker ™ Red dye for tracking of the SR were found co-localized. A quantitative analysis based on the intensity of different spectra demonstrated a local O 2 .− production derived from the SR. M 1 -receptor agonist, oxotremorine (Oxo) and a Ca 2+ ionophore, A23187 time-dependently increased this O 2 .− production colocalized with the SR. NOX inhibitors, diphenylene iodonium (DPI) and apocynin (Apo), or superoxide dismutase (SOD) and catalase, and Nox4 (a major intracellular NOX subunit) siRNA all substantially blocked this local production of O 2 .− , demonstrating an involvement of NOX. This SR-derived O 2 .− production was also abolished by the inhibitors of cyclic ADP-ribose (cADPR)-mediated Ca 2+ signaling, such as nicotinamide (Nicot, 6 mM), ryanodine (Rya, 50 μM) or 8-Br-cADPR (30 μM). However, IP 3 antagonist, 2-APB (50 μM) had no effect. In CAMs transfected with siRNA of ADP-ribosyl cyclase or RyR, this SR O 2 .− production was attenuated. Electron spin resonance (ESR) spectromic assay in purified SR also demonstrated the production of O 2 .− that was dependent on NOX activity and Ca 2+ concentrations. These results provide direct evidence that O 2 .− could be locally produced via NOX on the SR and that this local O 2 .− producing system is controlled by cADPR-RyR/Ca 2+ signaling pathway.

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Section: Discussionmentioning

confidence: 99%

Local production of O2− by NAD(P)H oxidase in the sarcoplasmic reticulum of coronary arterial myocytes: cADPR-mediated Ca2+ regulation

Zhang

Jin

Xia

et al. 2008

Cellular Signalling

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“…After enzymatic digestion of testicular tissue as mentioned before [14] with minor modifications, the reactive oxygen species (ROS) level was assayed by flow cytometry using 2 ′ , 7 ′ -dichlorofluorescin diacetate (DCFH-DA; Sigma, USA) according to previously mentioned method [31,13]. Briefly, appropriate amounts of testis homogenate (1-3 million cells) were incubated with DCFH-DA (10 mmol) for 15 min at 37°C with rotation in the dark to allow the probe to be incorporated into any membrane-bound vesicles.…”

Section: Assessment Of Reactive Oxygen Species In Testismentioning

confidence: 99%

TBHP-induced oxidative stress alters microRNAs expression in mouse testis

Fatemi

Sanati

Shamsara

et al. 2014

J Assist Reprod Genet

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Purpose Reactive oxygen species (ROS) and oxidative stress is one of the main reasons of male infertility. MicroRNAs (miRNAs) regulate multiple intracellular processes. Alterations in miRNAs expression may occur in different conditions and diseases. In this study, the effect of oxidative stress induced by tertiary-butyl hydroperoxide (TBHP) on the expression of candidate miRNAs in mouse testis was investigated.Methods After determining median lethal dose (LD 50 ), TBHP was intraperitoneally (ip) injected at the dilution of 1:10 LD 50 into the adult male mice for 2weeks, and then testis tissues were removed in order to assay the ROS level. Total RNA was extracted and the expression of five miRNAs was quantified by reverse transcription-real time polymerase chain reaction (RT-qPCR).Results The flow cytometry analysis showed a significant increase in ROS level in testis. The expression of three out of five selected miRNAs, including miR-34a, miR-181b and miR-122a, showed some degrees of changes following exposure to oxidative stress. These miRNAs are involved in antioxidant responses, inflammation pathway and spermatogenesis arrest. Conclusions In conclusion, TBHP alters the miRNA expression profile of testis which might play a potential role in oxidative and antioxidative responses and spermatogenesis.

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“…In 4-month-old SHR and W rats O2 −˙ production was lower in the presence of apocynin, but the reduction was not statistically significant. This may have been because the lucigenin method was unable to detect very slight differences in O2 −˙ levels that were only slightly above the background levels (25).…”

Section: Nad(p)h Oxidase Activity and O2mentioning

confidence: 99%

Is Cardiac Hypertrophy in Spontaneously Hypertensive Rats the Cause or the Consequence of Oxidative Stress?

et al. 2008

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The aim of this work was to assess the possible correlation between oxidative damage and the development of cardiac hypertrophy in heart tissue from young (40-d-old) and older (4-, 11-and 19-month-old) spontaneously hypertensive rats (SHR) in comparison with age-matched Wistar (W) rats. To this end, levels of thiobarbituric acid reactive substances (TBARS), nitrotyrosine contents, NAD(P)H oxidase activity, superoxide production, and the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were determined. Compared to age-matched normotensive rats, SHR showed a significant increase in systolic blood pressure from 40 d of age and left ventricular hypertrophy (LVH) was significantly evident from 4 months of age. W rats (11-and 19-month-old) also showed an increase in LVH with aging. TBARS and nitrotyrosine levels were similar in young rats from both strains and were significantly increased with age in both strains, with the values in SHR being significantly higher than those in age-matched W rats. NAD(P)H activity was similar in young SHR and W rats, whereas it was higher in aged SHR compared with age-matched W rats. Compared to W rats, superoxide production was higher in aged SHR, and was abolished by NAD(P)H inhibition with apocynin. CAT activity was increased in the hearts of 4-month-old SHR compared to age-matched W rats and was decreased in the hearts of the oldest SHR compared to the oldest W rats. SOD and GPx activities decreased in both rat strains with aging. Moreover, an increase in collagen deposition with aging was evident in both rat strains. Taken together, these data showed that aged SHR exhibited higher cardiac hypertrophy and oxidative damage compared to W rats, indicating that the two undesirable effects are associated. That is, oxidative stress appears to be a cause and/or consequence of hypertrophy development in this animal model. (Hypertens Res 2008; 31: 1465-1476)

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Measurement of Reactive Oxygen Species in Cardiovascular Studies

Cited by 465 publications

References 63 publications

Local production of O2− by NAD(P)H oxidase in the sarcoplasmic reticulum of coronary arterial myocytes: cADPR-mediated Ca2+ regulation

Local production of O2− by NAD(P)H oxidase in the sarcoplasmic reticulum of coronary arterial myocytes: cADPR-mediated Ca2+ regulation

TBHP-induced oxidative stress alters microRNAs expression in mouse testis

Is Cardiac Hypertrophy in Spontaneously Hypertensive Rats the Cause or the Consequence of Oxidative Stress?

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