- Email: [email protected]
Glutaredoxin-1 control on liver fibrosis in aged mice
activating type 2 immune responses. Contrasting the common belief that aging is associated with increased ROS
The proteasome plays a large role in maintaining protein
production, we observed that DUOX1 expression in mouse
homeostasis, the loss of which is a hallmark of aging.
lungs markedly decreases with age. Moreover, we noticed
Adaptive homeostasis, which is the transient modulation of
that DUOX1-deficient mice display accelerated age-related
the homeostatic range in response to non-damaging levels of
decline in lung function and features of emphysema, as
a stressor such as free radicals generated from oxidative
shown
lung
stress, is likewise diminished by aging. This adaptive
compliance. We hypothesized that loss of DUOX1 may
response, which has been clearly defined in many model
contribute to other features of lung aging, but aged-related
organisms and cell lines, has rarely been examined in
lung changes such as increased parenchymal collagen
primary cells, and never in Human Bronchial Epithelial cells
deposition, molecular indices of senescence, or tissue levels
(HBEs). These cells are continually exposed to higher
by
airspace
enlargement
and
increased
of pro-inflammatory cytokines, were all largely unaltered in
oxygen levels (relative to many other cell types) and offer
DUOX1-deficient mice. However, we observed that aged
an interesting perspective on the impact of chronic exposure
mice display markedly attenuated innate airway responses
to higher levels of oxygen on the adaptive response. Results
to inhaled allergens, as illustrated by diminished secretion
indicate HBEs from a young donor exhibit a strong adaptive
of IL-33 and other related cytokines. Similar findings were
response associated with the presence of the proteasome
observed in vitro using cultured mouse tracheal epithelial
regulator, Nrf2. Preliminary results also show a trend of
cells (MTEC), in which MTECs from older mice showed
diminishing adaptive response and Nrf2 levels from several
dramatically reduced innate responses to injury, which was
older donors. These results point to an interesting avenue
associated with reduced oxidation and activation of Src and
for future HBE studies investigating age and/or sex-related
EGFR. Consistent with a potential age-related suppression of
changes in adaptive responses in primary cells from males
DUOX1 by epigenetic mechanisms such as promoter
and females of different ages, with and without various
hypermethylation,
diseases.
allergen-induced
IL-33
secretion
in
MTECs from aged mice could be rescued by treatment with the DNA methyltransferase inhibitor AzdC. Overall, these
DOI: 10.1016/j.freeradbiomed.2017.10.078
results underline the importance of DUOX1 in airway host defense and regenerative capacity, and indicate that agerelated loss of DUOX1 contributes to accelerated impairment of
epithelial
regenerative
capacity
and
air-space
enlargement. Supported by Chiesi Pharmaceuticals
66 Glutaredoxin-1 control on liver fibrosis in aged mice
DOI: 10.1016/j.freeradbiomed.2017.10.077
Yuko Tsukahara1, Hlee Yang2, Beatriz Ferran Perez1, Markus M Bachschmid1, and Reiko Matsui1
65 Age-dependent Decline in Adaptive
1
Boston University School of Medicine, USA
2
Boston University, USA
Objective: Glutaredoxin-1 (Glrx) is a cytosolic enzyme
Homeostasis and Nrf2 Mediated Proteasome Induction in Human Bronchial Epithelial Cells Exposed to Oxidative
which removes glutathione (GSH) adducts of protein thiols (S-glutathionylation),
and
controls
protein
functions
including cellular signaling and gene transcription. We
Stress
previously showed that Glrx knockout middle-aged mice
Patrick Y. Sun1, Kelsi Yu1, and Kelvin J. A. Davies1
gene transfer suppressed lipid accumulation in the liver
1
University of Southern California, USA
developed liver steatosis and Glrx replenishment by viral (Shao 2016). It is reported that decreasing GSH adducts by inhibiting glutathione-S-transferase attenuates bleomycininduced lung fibrosis in mice. Therefore, we determine
SfRBM 2017
57
whether up-regulated Glrx can attenuate age-induced liver fibrosis and functional changes. Methods: Global Glrx transgenic (TG) and wild type (WT) mice of age 9-28 months were used to obtain liver sections to assess fibrotic areas by Picrosirius red collagen staining. Gene expression was assessed in the liver by RNA extraction and RT-qPCR. Hepatocytes and hepatic stellate cells were
were analyzed in plasma or whole blood: protein carbonyls, 3-nitrotyrosine,
malondialdehyde,
total
cysteine,
total
glutathione, β-carotene, lycopene, α-tocopherol, ascorbic acid and uric acid. All methods used were well-established in-house ELISA and HPLC methods, no commercial kits were used. Correlations were assessed as Pearson product
isolated to examine the effects of in vitro Glrx overexpression
moment correlation coefficients for transformed data. We observed statistically significant positive correlations
on collagen synthesis.
between age and MDA, cysteine, α-tocopherol as well as
Results: Our data showed that collagen staining in the liver
uric acid and negative associations with glutathione and
significantly increased in WT aged mice (> 18 months old) compared
to
WT
middle-aged
mice
(9
months).
Overexpression of Glrx in aged TG mice attenuated collagen deposition compared to WT aged mice. TGFβ mRNA expression in the liver was lower in TG than WT mice. Conclusion: Data indicate that aging increases liver fibrosis and Glrx overexpression suppresses the fibrotic change, in part, by inhibiting TGFβ expression. Further studies are required to examine signaling pathways and Glrx target(s) in aging liver. This study may demonstrate the protective role of Glrx and its anti-fibrotic effects in the liver.
lycopene. However, we found no association between age and protein carbonyls nor with 3-nitrotyrosine. Surprisingly, we observed an inverse association between protein carbonyls and MDA (r=-0.223, p<0.001) indicating that different redox biomarkers reflect different natures of the phenomenon “oxidative stress” and thus must be interpreted with caution. Since up to date no single biomarker is accepted as “gold standard” it is highly recommended to measure a combination of different biomarkers of oxidative stress. In conclusion, the oxidative stress status of a person depends upon factors such as age, lifestyle and genetics and last but not least also on the biomarker itself and the methods used.
DOI: 10.1016/j.freeradbiomed.2017.10.079
DOI: 10.1016/j.freeradbiomed.2017.10.080
67 Oxidative stress biomarkers in the MARKAGE Study Daniela Weber1, Wolfgang Stuetz2, Alexander Buerkle3,4, and Tilman Grune1 1
German Institute of Human Nutrition, Germany
2
University of Hohenheim, Germany
3
University of Konstanz, Germany
4
on behalf of the MARK-AGE Consortium, Germany
20S Proteasome Beta 5 Subunit is Crucial for Sexually Divergent Adaptive Homeostasis Responses to Oxidative Stress in D. melanogaster Christina Sisliyan1, Sarah Wong1, Laura Corrales-Diaz Pomatto1, John Tower1, and Kelvin J. A. Davies1
Since the introduction of the term “oxidative stress” in 1985 this concept has been in the spotlight of research due to its association with age and different diseases. Previous human studies on oxidative stress and aging have focused mostly on single biomarkers, thus making it difficult to compare results and establish reference values. The aim of this study was to assess different redox biomarkers in participants of the MARK-AGE project. Healthy participants aged 35-75 years from eight European countries were recruited in an age-stratified manner (n=2,207). The following biomarkers
58
68
1
University of Southern California, USA
The 20S proteasome, which exhibits chymotrypsin-like, trypsin-like,
and
caspase-like
proteolytic
activities,
is
upregulated during adaptation to oxidative stress and provides significant protection against toxicity otherwise exerted by aggregated and cross-linked protein aggregates. We have previously shown a female-specific adaptation to hydrogen peroxide (H2O2) measured by changes in 20S
proteasome expression, proteolytic activity, and survival
SfRBM 2017
The proteasome plays a large role in maintaining protein
production, we observed that DUOX1 expression in mouse
homeostasis, the loss of which is a hallmark of aging.
lungs markedly decreases with age. Moreover, we noticed
Adaptive homeostasis, which is the transient modulation of
that DUOX1-deficient mice display accelerated age-related
the homeostatic range in response to non-damaging levels of
decline in lung function and features of emphysema, as
a stressor such as free radicals generated from oxidative
shown
lung
stress, is likewise diminished by aging. This adaptive
compliance. We hypothesized that loss of DUOX1 may
response, which has been clearly defined in many model
contribute to other features of lung aging, but aged-related
organisms and cell lines, has rarely been examined in
lung changes such as increased parenchymal collagen
primary cells, and never in Human Bronchial Epithelial cells
deposition, molecular indices of senescence, or tissue levels
(HBEs). These cells are continually exposed to higher
by
airspace
enlargement
and
increased
of pro-inflammatory cytokines, were all largely unaltered in
oxygen levels (relative to many other cell types) and offer
DUOX1-deficient mice. However, we observed that aged
an interesting perspective on the impact of chronic exposure
mice display markedly attenuated innate airway responses
to higher levels of oxygen on the adaptive response. Results
to inhaled allergens, as illustrated by diminished secretion
indicate HBEs from a young donor exhibit a strong adaptive
of IL-33 and other related cytokines. Similar findings were
response associated with the presence of the proteasome
observed in vitro using cultured mouse tracheal epithelial
regulator, Nrf2. Preliminary results also show a trend of
cells (MTEC), in which MTECs from older mice showed
diminishing adaptive response and Nrf2 levels from several
dramatically reduced innate responses to injury, which was
older donors. These results point to an interesting avenue
associated with reduced oxidation and activation of Src and
for future HBE studies investigating age and/or sex-related
EGFR. Consistent with a potential age-related suppression of
changes in adaptive responses in primary cells from males
DUOX1 by epigenetic mechanisms such as promoter
and females of different ages, with and without various
hypermethylation,
diseases.
allergen-induced
IL-33
secretion
in
MTECs from aged mice could be rescued by treatment with the DNA methyltransferase inhibitor AzdC. Overall, these
DOI: 10.1016/j.freeradbiomed.2017.10.078
results underline the importance of DUOX1 in airway host defense and regenerative capacity, and indicate that agerelated loss of DUOX1 contributes to accelerated impairment of
epithelial
regenerative
capacity
and
air-space
enlargement. Supported by Chiesi Pharmaceuticals
66 Glutaredoxin-1 control on liver fibrosis in aged mice
DOI: 10.1016/j.freeradbiomed.2017.10.077
Yuko Tsukahara1, Hlee Yang2, Beatriz Ferran Perez1, Markus M Bachschmid1, and Reiko Matsui1
65 Age-dependent Decline in Adaptive
1
Boston University School of Medicine, USA
2
Boston University, USA
Objective: Glutaredoxin-1 (Glrx) is a cytosolic enzyme
Homeostasis and Nrf2 Mediated Proteasome Induction in Human Bronchial Epithelial Cells Exposed to Oxidative
which removes glutathione (GSH) adducts of protein thiols (S-glutathionylation),
and
controls
protein
functions
including cellular signaling and gene transcription. We
Stress
previously showed that Glrx knockout middle-aged mice
Patrick Y. Sun1, Kelsi Yu1, and Kelvin J. A. Davies1
gene transfer suppressed lipid accumulation in the liver
1
University of Southern California, USA
developed liver steatosis and Glrx replenishment by viral (Shao 2016). It is reported that decreasing GSH adducts by inhibiting glutathione-S-transferase attenuates bleomycininduced lung fibrosis in mice. Therefore, we determine
SfRBM 2017
57
whether up-regulated Glrx can attenuate age-induced liver fibrosis and functional changes. Methods: Global Glrx transgenic (TG) and wild type (WT) mice of age 9-28 months were used to obtain liver sections to assess fibrotic areas by Picrosirius red collagen staining. Gene expression was assessed in the liver by RNA extraction and RT-qPCR. Hepatocytes and hepatic stellate cells were
were analyzed in plasma or whole blood: protein carbonyls, 3-nitrotyrosine,
malondialdehyde,
total
cysteine,
total
glutathione, β-carotene, lycopene, α-tocopherol, ascorbic acid and uric acid. All methods used were well-established in-house ELISA and HPLC methods, no commercial kits were used. Correlations were assessed as Pearson product
isolated to examine the effects of in vitro Glrx overexpression
moment correlation coefficients for transformed data. We observed statistically significant positive correlations
on collagen synthesis.
between age and MDA, cysteine, α-tocopherol as well as
Results: Our data showed that collagen staining in the liver
uric acid and negative associations with glutathione and
significantly increased in WT aged mice (> 18 months old) compared
to
WT
middle-aged
mice
(9
months).
Overexpression of Glrx in aged TG mice attenuated collagen deposition compared to WT aged mice. TGFβ mRNA expression in the liver was lower in TG than WT mice. Conclusion: Data indicate that aging increases liver fibrosis and Glrx overexpression suppresses the fibrotic change, in part, by inhibiting TGFβ expression. Further studies are required to examine signaling pathways and Glrx target(s) in aging liver. This study may demonstrate the protective role of Glrx and its anti-fibrotic effects in the liver.
lycopene. However, we found no association between age and protein carbonyls nor with 3-nitrotyrosine. Surprisingly, we observed an inverse association between protein carbonyls and MDA (r=-0.223, p<0.001) indicating that different redox biomarkers reflect different natures of the phenomenon “oxidative stress” and thus must be interpreted with caution. Since up to date no single biomarker is accepted as “gold standard” it is highly recommended to measure a combination of different biomarkers of oxidative stress. In conclusion, the oxidative stress status of a person depends upon factors such as age, lifestyle and genetics and last but not least also on the biomarker itself and the methods used.
DOI: 10.1016/j.freeradbiomed.2017.10.079
DOI: 10.1016/j.freeradbiomed.2017.10.080
67 Oxidative stress biomarkers in the MARKAGE Study Daniela Weber1, Wolfgang Stuetz2, Alexander Buerkle3,4, and Tilman Grune1 1
German Institute of Human Nutrition, Germany
2
University of Hohenheim, Germany
3
University of Konstanz, Germany
4
on behalf of the MARK-AGE Consortium, Germany
20S Proteasome Beta 5 Subunit is Crucial for Sexually Divergent Adaptive Homeostasis Responses to Oxidative Stress in D. melanogaster Christina Sisliyan1, Sarah Wong1, Laura Corrales-Diaz Pomatto1, John Tower1, and Kelvin J. A. Davies1
Since the introduction of the term “oxidative stress” in 1985 this concept has been in the spotlight of research due to its association with age and different diseases. Previous human studies on oxidative stress and aging have focused mostly on single biomarkers, thus making it difficult to compare results and establish reference values. The aim of this study was to assess different redox biomarkers in participants of the MARK-AGE project. Healthy participants aged 35-75 years from eight European countries were recruited in an age-stratified manner (n=2,207). The following biomarkers
58
68
1
University of Southern California, USA
The 20S proteasome, which exhibits chymotrypsin-like, trypsin-like,
and
caspase-like
proteolytic
activities,
is
upregulated during adaptation to oxidative stress and provides significant protection against toxicity otherwise exerted by aggregated and cross-linked protein aggregates. We have previously shown a female-specific adaptation to hydrogen peroxide (H2O2) measured by changes in 20S
proteasome expression, proteolytic activity, and survival
SfRBM 2017