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Perinatal tobacco smoke exposure increases vascular oxidative stress and mitochondrial damage in non-human primates.

Westbrook DG, Anderson PG, Pinkerton KE, Ballinger SW - Cardiovasc. Toxicol. (2010)

Bottom Line: Epidemiological studies suggest that events occurring during fetal and early childhood development influence disease susceptibility.M. mulatta were exposed to low levels of ETS (1 mg/m(3) total suspended particulates) from gestation (day 40) to early childhood (1 year), and aortic tissues were assessed for oxidized proteins (protein carbonyls), antioxidant activity (SOD), mitochondrial function (cytochrome oxidase), and mitochondrial damage (mitochondrial DNA damage).Results revealed that perinatal ETS exposure resulted in significantly increased oxidative stress, mitochondrial dysfunction and damage which were accompanied by significantly decreased mitochondrial antioxidant capacity and mitochondrial copy number in vascular tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Division of Molecular and Cellular Pathology, University of Alabama at Birmingham, VH G019F, 1530 3rd Avenue S., Birmingham, AL 35294-0019, USA.

ABSTRACT
Epidemiological studies suggest that events occurring during fetal and early childhood development influence disease susceptibility. Similarly, molecular studies in mice have shown that in utero exposure to cardiovascular disease (CVD) risk factors such as environmental tobacco smoke (ETS) increased adult atherogenic susceptibility and mitochondrial damage; however, the molecular effects of similar exposures in primates are not yet known. To determine whether perinatal ETS exposure increased mitochondrial damage, dysfunction and oxidant stress in primates, archived tissues from the non-human primate model Macaca mulatta (M. mulatta) were utilized. M. mulatta were exposed to low levels of ETS (1 mg/m(3) total suspended particulates) from gestation (day 40) to early childhood (1 year), and aortic tissues were assessed for oxidized proteins (protein carbonyls), antioxidant activity (SOD), mitochondrial function (cytochrome oxidase), and mitochondrial damage (mitochondrial DNA damage). Results revealed that perinatal ETS exposure resulted in significantly increased oxidative stress, mitochondrial dysfunction and damage which were accompanied by significantly decreased mitochondrial antioxidant capacity and mitochondrial copy number in vascular tissue. Increased mitochondrial damage was also detected in buffy coat tissues in exposed M. mulatta. These studies suggest that perinatal tobacco smoke exposure increases vascular oxidative stress and mitochondrial damage in primates, potentially increasing adult disease susceptibility.

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Mitochondrial DNA (mtDNA) damage in M. mulatta aorta. MtDNA damage was quantified from genomic DNA preparations extracted from aorta. a mtDNA damage determined by QPCR; (n = 3/group/tissue). The inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the full-length QPCR product (Long) that is used to quantify relative levels of mtDNA damage (less product indicates increased damage) relative to control, whereas the lower inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the smaller QPCR product (Short) that is used for mtDNA copy number normalization. Bar graph shows the relative level of mtDNA damage (normalized for copy number) between exposure groups. b mtDNA damage detected by Fpg digestion followed by QPCR. The inset shows QPCR results for Fpg-treated unexposed controls and ETS-exposed animals (lanes 1–3 and 4–6, respectively), showing the full-length QPCR product (less product indicates increased mtDNA damage). The bar graph shows the difference in Fpg detectable lesions (±Fpg) in ETS-exposed monkeys relative to unexposed controls. Unexposed, M. mulatta age-matched control; ETS, M. mulatta exposed to 1 mg/m3 ETS during gestation to 1 year of age. Asterisks indicate significant difference (P ≤ 0.05) from unexposed M. mulatta
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Fig3: Mitochondrial DNA (mtDNA) damage in M. mulatta aorta. MtDNA damage was quantified from genomic DNA preparations extracted from aorta. a mtDNA damage determined by QPCR; (n = 3/group/tissue). The inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the full-length QPCR product (Long) that is used to quantify relative levels of mtDNA damage (less product indicates increased damage) relative to control, whereas the lower inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the smaller QPCR product (Short) that is used for mtDNA copy number normalization. Bar graph shows the relative level of mtDNA damage (normalized for copy number) between exposure groups. b mtDNA damage detected by Fpg digestion followed by QPCR. The inset shows QPCR results for Fpg-treated unexposed controls and ETS-exposed animals (lanes 1–3 and 4–6, respectively), showing the full-length QPCR product (less product indicates increased mtDNA damage). The bar graph shows the difference in Fpg detectable lesions (±Fpg) in ETS-exposed monkeys relative to unexposed controls. Unexposed, M. mulatta age-matched control; ETS, M. mulatta exposed to 1 mg/m3 ETS during gestation to 1 year of age. Asterisks indicate significant difference (P ≤ 0.05) from unexposed M. mulatta

Mentions: To determine whether perinatal ETS exposure impacted mitochondrial integrity, quantitative PCR was performed on aortic tissue collected from M. mulatta. Figure 3 illustrates that perinatal ETS exposure resulted in significantly increased levels of mtDNA damage in aortic tissues compared to controls (Fig. 3a; less product in the “long” row on inset reflects more mtDNA damage). To investigate whether an additional measure of DNA damage yielded differences between unexposed and ETS-exposed animals, aortic DNA samples were also treated with formamidopyrimidine (fapy) DNA glycosylase (Fpg), which cleaves DNA at specific lesions such as 7, 8-dihydro-8-oxoguanonine, 8-oxoadenine, fapy-guanine, methy-fapy-guanine, fapy-adenine and 5-hydroxy-cytosine. Figure 3b shows that Fpg treatment revealed net increased levels of mtDNA lesions in exposed animals compared to unexposed controls (less product on inset reflects more mtDNA damage).Fig. 3


Perinatal tobacco smoke exposure increases vascular oxidative stress and mitochondrial damage in non-human primates.

Westbrook DG, Anderson PG, Pinkerton KE, Ballinger SW - Cardiovasc. Toxicol. (2010)

Mitochondrial DNA (mtDNA) damage in M. mulatta aorta. MtDNA damage was quantified from genomic DNA preparations extracted from aorta. a mtDNA damage determined by QPCR; (n = 3/group/tissue). The inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the full-length QPCR product (Long) that is used to quantify relative levels of mtDNA damage (less product indicates increased damage) relative to control, whereas the lower inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the smaller QPCR product (Short) that is used for mtDNA copy number normalization. Bar graph shows the relative level of mtDNA damage (normalized for copy number) between exposure groups. b mtDNA damage detected by Fpg digestion followed by QPCR. The inset shows QPCR results for Fpg-treated unexposed controls and ETS-exposed animals (lanes 1–3 and 4–6, respectively), showing the full-length QPCR product (less product indicates increased mtDNA damage). The bar graph shows the difference in Fpg detectable lesions (±Fpg) in ETS-exposed monkeys relative to unexposed controls. Unexposed, M. mulatta age-matched control; ETS, M. mulatta exposed to 1 mg/m3 ETS during gestation to 1 year of age. Asterisks indicate significant difference (P ≤ 0.05) from unexposed M. mulatta
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Related In: Results  -  Collection

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Fig3: Mitochondrial DNA (mtDNA) damage in M. mulatta aorta. MtDNA damage was quantified from genomic DNA preparations extracted from aorta. a mtDNA damage determined by QPCR; (n = 3/group/tissue). The inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the full-length QPCR product (Long) that is used to quantify relative levels of mtDNA damage (less product indicates increased damage) relative to control, whereas the lower inset (lanes 1–3 are unexposed control, lanes 4–6 are ETS exposed) shows the smaller QPCR product (Short) that is used for mtDNA copy number normalization. Bar graph shows the relative level of mtDNA damage (normalized for copy number) between exposure groups. b mtDNA damage detected by Fpg digestion followed by QPCR. The inset shows QPCR results for Fpg-treated unexposed controls and ETS-exposed animals (lanes 1–3 and 4–6, respectively), showing the full-length QPCR product (less product indicates increased mtDNA damage). The bar graph shows the difference in Fpg detectable lesions (±Fpg) in ETS-exposed monkeys relative to unexposed controls. Unexposed, M. mulatta age-matched control; ETS, M. mulatta exposed to 1 mg/m3 ETS during gestation to 1 year of age. Asterisks indicate significant difference (P ≤ 0.05) from unexposed M. mulatta
Mentions: To determine whether perinatal ETS exposure impacted mitochondrial integrity, quantitative PCR was performed on aortic tissue collected from M. mulatta. Figure 3 illustrates that perinatal ETS exposure resulted in significantly increased levels of mtDNA damage in aortic tissues compared to controls (Fig. 3a; less product in the “long” row on inset reflects more mtDNA damage). To investigate whether an additional measure of DNA damage yielded differences between unexposed and ETS-exposed animals, aortic DNA samples were also treated with formamidopyrimidine (fapy) DNA glycosylase (Fpg), which cleaves DNA at specific lesions such as 7, 8-dihydro-8-oxoguanonine, 8-oxoadenine, fapy-guanine, methy-fapy-guanine, fapy-adenine and 5-hydroxy-cytosine. Figure 3b shows that Fpg treatment revealed net increased levels of mtDNA lesions in exposed animals compared to unexposed controls (less product on inset reflects more mtDNA damage).Fig. 3

Bottom Line: Epidemiological studies suggest that events occurring during fetal and early childhood development influence disease susceptibility.M. mulatta were exposed to low levels of ETS (1 mg/m(3) total suspended particulates) from gestation (day 40) to early childhood (1 year), and aortic tissues were assessed for oxidized proteins (protein carbonyls), antioxidant activity (SOD), mitochondrial function (cytochrome oxidase), and mitochondrial damage (mitochondrial DNA damage).Results revealed that perinatal ETS exposure resulted in significantly increased oxidative stress, mitochondrial dysfunction and damage which were accompanied by significantly decreased mitochondrial antioxidant capacity and mitochondrial copy number in vascular tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Division of Molecular and Cellular Pathology, University of Alabama at Birmingham, VH G019F, 1530 3rd Avenue S., Birmingham, AL 35294-0019, USA.

ABSTRACT
Epidemiological studies suggest that events occurring during fetal and early childhood development influence disease susceptibility. Similarly, molecular studies in mice have shown that in utero exposure to cardiovascular disease (CVD) risk factors such as environmental tobacco smoke (ETS) increased adult atherogenic susceptibility and mitochondrial damage; however, the molecular effects of similar exposures in primates are not yet known. To determine whether perinatal ETS exposure increased mitochondrial damage, dysfunction and oxidant stress in primates, archived tissues from the non-human primate model Macaca mulatta (M. mulatta) were utilized. M. mulatta were exposed to low levels of ETS (1 mg/m(3) total suspended particulates) from gestation (day 40) to early childhood (1 year), and aortic tissues were assessed for oxidized proteins (protein carbonyls), antioxidant activity (SOD), mitochondrial function (cytochrome oxidase), and mitochondrial damage (mitochondrial DNA damage). Results revealed that perinatal ETS exposure resulted in significantly increased oxidative stress, mitochondrial dysfunction and damage which were accompanied by significantly decreased mitochondrial antioxidant capacity and mitochondrial copy number in vascular tissue. Increased mitochondrial damage was also detected in buffy coat tissues in exposed M. mulatta. These studies suggest that perinatal tobacco smoke exposure increases vascular oxidative stress and mitochondrial damage in primates, potentially increasing adult disease susceptibility.

Show MeSH
Related in: MedlinePlus