Abstract 16844: Relative Endothelial Toxicity of Tobacco Smoke and E-Cigarette Aerosol: A Functional and Mechanistic Assessment
BackgroundSmoking cigarettes decreases expression of eNOS in the endothelium, resulting in lower nitric oxide (NO) secretion and decreased flow-mediated dilation in the conducting arteries. In contrast, the effects of e-cigarettes (e-cigs) on endothelial function are just beginning to be studied and...
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Published in | Circulation (New York, N.Y.) Vol. 138; no. Suppl_1 Suppl 1; p. A16844 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
by the American College of Cardiology Foundation and the American Heart Association, Inc
06.11.2018
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Abstract | BackgroundSmoking cigarettes decreases expression of eNOS in the endothelium, resulting in lower nitric oxide (NO) secretion and decreased flow-mediated dilation in the conducting arteries. In contrast, the effects of e-cigarettes (e-cigs) on endothelial function are just beginning to be studied and the mechanisms of action are unclear.AimTo test the hypothesis that circulating factors from e-cigarette users decrease endothelial eNOS protein levels and NO secretion in primary endothelial cell cultures, relative to nonsmokers and cigarette smokers.Methods36 healthy individuals were recruited and grouped as nonsmokers (n=12), cigarette smokers (n=7), and e-cig users (n=17). Human umbilical vein endothelial cells (HUVECs) were cultured at 20,000 cells per well in 24-well culture plate. Serum samples from individual subjects and endothelial growth media were added at a 1:1 ratio to confluent cells and incubated for 12 hours (basal condition), followed by fresh medium containing VEGF at 50 ng/ml for 30 min (stimulated condition). The amount of NO liberated from cells was measured in culture supernatants by the chemilumin- escence method using a NO analyzer. Endothelial NO synthase (eNOS) protein levels in HUVECs was measured in cell lysates by ELISA. NO and eNOS results were normalized to cell number.ResultsThe cells treated with serum from e-cig users vs. nonsmokers produced less NO upon stimulation (P<.03) and contained less eNOS protein (P<.03). eNOS protein level was lower in the e-cig user serum group than in the cigarette smoker serum group (P<.05), although stimulated NO production was decreased less by e-cig user serum than by smoker serum. Lower eNOS levels in the cigarette group vs. non-smoker group was not significant, but stimulated NO was significantly lower in the cigarette group vs. nonsmoker (P<.006; see figure).ConclusionExposure of cultured endothelial cells to circulating factors from e-cig users, relative to non-users, leads to lower eNOS protein levels and decreased NO production. |
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AbstractList | BackgroundSmoking cigarettes decreases expression of eNOS in the endothelium, resulting in lower nitric oxide (NO) secretion and decreased flow-mediated dilation in the conducting arteries. In contrast, the effects of e-cigarettes (e-cigs) on endothelial function are just beginning to be studied and the mechanisms of action are unclear.AimTo test the hypothesis that circulating factors from e-cigarette users decrease endothelial eNOS protein levels and NO secretion in primary endothelial cell cultures, relative to nonsmokers and cigarette smokers.Methods36 healthy individuals were recruited and grouped as nonsmokers (n=12), cigarette smokers (n=7), and e-cig users (n=17). Human umbilical vein endothelial cells (HUVECs) were cultured at 20,000 cells per well in 24-well culture plate. Serum samples from individual subjects and endothelial growth media were added at a 1:1 ratio to confluent cells and incubated for 12 hours (basal condition), followed by fresh medium containing VEGF at 50 ng/ml for 30 min (stimulated condition). The amount of NO liberated from cells was measured in culture supernatants by the chemilumin- escence method using a NO analyzer. Endothelial NO synthase (eNOS) protein levels in HUVECs was measured in cell lysates by ELISA. NO and eNOS results were normalized to cell number.ResultsThe cells treated with serum from e-cig users vs. nonsmokers produced less NO upon stimulation (P<.03) and contained less eNOS protein (P<.03). eNOS protein level was lower in the e-cig user serum group than in the cigarette smoker serum group (P<.05), although stimulated NO production was decreased less by e-cig user serum than by smoker serum. Lower eNOS levels in the cigarette group vs. non-smoker group was not significant, but stimulated NO was significantly lower in the cigarette group vs. nonsmoker (P<.006; see figure).ConclusionExposure of cultured endothelial cells to circulating factors from e-cig users, relative to non-users, leads to lower eNOS protein levels and decreased NO production. Abstract only Background: Smoking cigarettes decreases expression of eNOS in the endothelium, resulting in lower nitric oxide (NO) secretion and decreased flow-mediated dilation in the conducting arteries. In contrast, the effects of e-cigarettes (e-cigs) on endothelial function are just beginning to be studied and the mechanisms of action are unclear. Aim: To test the hypothesis that circulating factors from e-cigarette users decrease endothelial eNOS protein levels and NO secretion in primary endothelial cell cultures, relative to nonsmokers and cigarette smokers. Methods: 36 healthy individuals were recruited and grouped as nonsmokers (n=12), cigarette smokers (n=7), and e-cig users (n=17). Human umbilical vein endothelial cells (HUVECs) were cultured at 20,000 cells per well in 24-well culture plate. Serum samples from individual subjects and endothelial growth media were added at a 1:1 ratio to confluent cells and incubated for 12 hours (basal condition), followed by fresh medium containing VEGF at 50 ng/ml for 30 min (stimulated condition). The amount of NO liberated from cells was measured in culture supernatants by the chemilumin- escence method using a NO analyzer. Endothelial NO synthase (eNOS) protein levels in HUVECs was measured in cell lysates by ELISA. NO and eNOS results were normalized to cell number. Results: The cells treated with serum from e-cig users vs. nonsmokers produced less NO upon stimulation (P<.03) and contained less eNOS protein (P<.03). eNOS protein level was lower in the e-cig user serum group than in the cigarette smoker serum group (P<.05), although stimulated NO production was decreased less by e-cig user serum than by smoker serum. Lower eNOS levels in the cigarette group vs. non-smoker group was not significant, but stimulated NO was significantly lower in the cigarette group vs. nonsmoker (P<.006; see figure). Conclusion: Exposure of cultured endothelial cells to circulating factors from e-cig users, relative to non-users, leads to lower eNOS protein levels and decreased NO production. |
Author | Huang, Abel Whitlatch, Adam Han, Daniel D Springer, Matthew L Mohammadi, Leila Schick, Suzaynn F Derakhshandeh, Ronak |
AuthorAffiliation | Cardiovascular Rsch Institute, UCSF, San Francisco, CA Medicine, UCSF, San Francisco, CA Cardiology, UCSF, San Francisco, CA |
AuthorAffiliation_xml | – name: Cardiovascular Rsch Institute, UCSF, San Francisco, CA – name: Cardiology, UCSF, San Francisco, CA – name: Medicine, UCSF, San Francisco, CA |
Author_xml | – sequence: 1 givenname: Leila surname: Mohammadi fullname: Mohammadi, Leila organization: Cardiovascular Rsch Institute, UCSF, San Francisco, CA – sequence: 2 givenname: Ronak surname: Derakhshandeh fullname: Derakhshandeh, Ronak organization: Cardiovascular Rsch Institute, UCSF, San Francisco, CA – sequence: 3 givenname: Daniel surname: Han middlename: D fullname: Han, Daniel D organization: Cardiology, UCSF, San Francisco, CA – sequence: 4 givenname: Adam surname: Whitlatch fullname: Whitlatch, Adam organization: Medicine, UCSF, San Francisco, CA – sequence: 5 givenname: Abel surname: Huang fullname: Huang, Abel organization: Medicine, UCSF, San Francisco, CA – sequence: 6 givenname: Suzaynn surname: Schick middlename: F fullname: Schick, Suzaynn F organization: Medicine, UCSF, San Francisco, CA – sequence: 7 givenname: Matthew surname: Springer middlename: L fullname: Springer, Matthew L organization: Cardiology, UCSF, San Francisco, CA |
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Snippet | BackgroundSmoking cigarettes decreases expression of eNOS in the endothelium, resulting in lower nitric oxide (NO) secretion and decreased flow-mediated... Abstract only Background: Smoking cigarettes decreases expression of eNOS in the endothelium, resulting in lower nitric oxide (NO) secretion and decreased... |
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Title | Abstract 16844: Relative Endothelial Toxicity of Tobacco Smoke and E-Cigarette Aerosol: A Functional and Mechanistic Assessment |
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