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_:vb419250985 _:vb419250986 _:vb419250987 _:vb419251216 _:vb419251217 _:vb419251204 _:vb419251205 _:vb419251206 _:vb419251207 _:vb419251200 _:vb419251201 _:vb419251202 _:vb419251203 _:vb419251212 _:vb419251213 _:vb419251214 _:vb419251215 _:vb419251208 _:vb419251209 _:vb419251210 _:vb419251211
n3:pmcid
PMC0
bibo:doi
10.2337%2Fdb09-0482
n7:contains
_:vb6461021 _:vb6461047 _:vb6461073
Subject Item
_:vb6461021
rdf:type
n7:Section
dc:title
results
n7:contains
_:vb6461022 _:vb6461023 _:vb6461040 _:vb6461041 _:vb6461042 _:vb6461043 _:vb6461044 _:vb6461045 _:vb6461046 _:vb6461024 _:vb6461025 _:vb6461026 _:vb6461027 _:vb6461028 _:vb6461029 _:vb6461030 _:vb6461031 _:vb6461032 _:vb6461033 _:vb6461034 _:vb6461035 _:vb6461036 _:vb6461037 _:vb6461038 _:vb6461039
Subject Item
_:vb6461022
rdf:type
n3:Context
rdf:value
As reported previously (>>7<<), resveratrol increased physical endurance (309.2 ± 37 vs. 212.3 ± 19 m) (Fig. 3A) of wild-type mice.
n3:mentions
n2:17112576
Subject Item
_:vb6461023
rdf:type
n3:Context
rdf:value
Prolonged activation of AMPK increases Glut4 expression (>>33<<). To determine whether increased expression of Glut4 expression was involved in the resveratrol-mediated improvement of physical endurance, we measured Glut4 mRNA levels in skeletal muscle.
n3:mentions
n2:10562646
Subject Item
_:vb6461024
rdf:type
n3:Context
rdf:value
It is believed that the resveratrol effects are mediated by increased expression and function of PGC-1α, the master regulator of mitochondrial biogenesis and oxidative phosphorylation (>>34<<). We measured the expression of PGC-1α and other resveratrol-induced genes critical for mitochondrial function in the skeletal muscle of resveratrol-treated AMPK-deficient mice. As shown in Fig. 3C, resveratrol induced the expression of
n3:mentions
n2:9529258
Subject Item
_:vb6461025
rdf:type
n3:Context
rdf:value
To determine whether the requirement of AMPK for the resveratrol effect is cell autonomous, we treated mefs developed from AMPKα1/α2 double knockout (>>32<<) and wild-type embryos with resveratrol for 6–24 h.
n3:mentions
n2:16809770
Subject Item
_:vb6461026
rdf:type
n3:Context
rdf:value
Sirt1 overexpression (>>35<<–37), AMPK activation (21,22,38), and resveratrol treatment (5,7) all improve glucose tolerance and insulin sensitivity.
n3:mentions
n2:17877786 n2:18599449 n2:18840364
Subject Item
_:vb6461027
rdf:type
n3:Context
rdf:value
Sirt1 overexpression (35–37), AMPK activation (>>21<<,22,38), and resveratrol treatment (5,7) all improve glucose tolerance and insulin sensitivity.
n3:mentions
n2:18674809
Subject Item
_:vb6461028
rdf:type
n3:Context
rdf:value
Sirt1 overexpression (35–37), AMPK activation (21,>>22<<,38), and resveratrol treatment (5,7) all improve glucose tolerance and insulin sensitivity.
n3:mentions
n2:12511592
Subject Item
_:vb6461029
rdf:type
n3:Context
rdf:value
Sirt1 overexpression (35–37), AMPK activation (21,22,>>38<<), and resveratrol treatment (5,7) all improve glucose tolerance and insulin sensitivity.
n3:mentions
n2:12368907
Subject Item
_:vb6461030
rdf:type
n3:Context
rdf:value
Sirt1 overexpression (35–37), AMPK activation (21,22,38), and resveratrol treatment (>>5<<,7) all improve glucose tolerance and insulin sensitivity.
n3:mentions
n2:17086191
Subject Item
_:vb6461031
rdf:type
n3:Context
rdf:value
Sirt1 overexpression (35–37), AMPK activation (21,22,38), and resveratrol treatment (5,>>7<<) all improve glucose tolerance and insulin sensitivity.
n3:mentions
n2:17112576
Subject Item
_:vb6461032
rdf:type
n3:Context
rdf:value
C: Relative levels of ROS in skeletal muscle extracts from wild-type (WT) and AMPKα1−/− mice fed high-fat diet (□) alone or high-fat diet supplemented with resveratrol (■) as measured by DCF fluorescence (>>52<<) (n = 4–5 for each genotype).
n3:mentions
n2:16847346
Subject Item
_:vb6461033
rdf:type
n3:Context
rdf:value
Insulin-resistant states are associated with increased production of ROS, and reduction of ROS with antioxidants improves insulin sensitivity (>>39<<,40). Since resveratrol is thought to have antioxidant properties, it is possible that some insulin sensitizing of resveratrol effect may be related to its antioxidant effect.
n3:mentions
n2:16612386
Subject Item
_:vb6461034
rdf:type
n3:Context
rdf:value
Insulin-resistant states are associated with increased production of ROS, and reduction of ROS with antioxidants improves insulin sensitivity (39,>>40<<). Since resveratrol is thought to have antioxidant properties, it is possible that some insulin sensitizing of resveratrol effect may be related to its antioxidant effect.
n3:mentions
n2:17130651
Subject Item
_:vb6461035
rdf:type
n3:Context
rdf:value
Insulin-resistant tissues are often associated with elevated levels of intramyocellular lipids such as diacylglyceride (DAG) and ceramide (>>41<<). Mitochondrial dysfunction decreases fat oxidation, leading to accumulation of these lipids, which decrease insulin sensitivity (41,42).
n3:mentions
n2:2186032
Subject Item
_:vb6461036
rdf:type
n3:Context
rdf:value
Mitochondrial dysfunction decreases fat oxidation, leading to accumulation of these lipids, which decrease insulin sensitivity (>>41<<,42). Since resveratrol increases mitochondrial function (7), we measured the DAG and ceramide levels in resveratrol-treated skeletal muscle. As shown in Fig. 4D and E, resveratrol decreased DAG and ceramide levels in wild-type mice but
n3:mentions
n2:2186032
Subject Item
_:vb6461037
rdf:type
n3:Context
rdf:value
Mitochondrial dysfunction decreases fat oxidation, leading to accumulation of these lipids, which decrease insulin sensitivity (41,>>42<<). Since resveratrol increases mitochondrial function (7), we measured the DAG and ceramide levels in resveratrol-treated skeletal muscle. As shown in Fig. 4D and E, resveratrol decreased DAG and ceramide levels in wild-type mice but not
n3:mentions
n2:9710629
Subject Item
_:vb6461038
rdf:type
n3:Context
rdf:value
Since resveratrol increases mitochondrial function (>>7<<), we measured the DAG and ceramide levels in resveratrol-treated skeletal muscle.
n3:mentions
n2:17112576
Subject Item
_:vb6461039
rdf:type
n3:Context
rdf:value
Adiponectin, a cytokine that is produced by fat and is decreased in obese and diabetic subjects (>>43<<), increases insulin sensitivity by activating AMPK (38).
n3:mentions
n2:8631877
Subject Item
_:vb6461040
rdf:type
n3:Context
rdf:value
Adiponectin, a cytokine that is produced by fat and is decreased in obese and diabetic subjects (43), increases insulin sensitivity by activating AMPK (>>38<<). To determine whether resveratrol increased insulin sensitivity by increasing adiponectin levels, we measured serum adiponectin levels in resveratrol-treated mice. Adiponectin levels were similar for wild-type, AMPKα1−/−, and AMPKα2−/−
n3:mentions
n2:12368907
Subject Item
_:vb6461041
rdf:type
n3:Context
rdf:value
Although this can be explained, at least in part, by the observation that resveratrol inhibits insulin secretion by inhibiting the electrical activity in the β-cells (>>44<<), more studies are required to fully understand the disparity between the insulin levels and insulin sensitivity in resveratrol-treated AMPK-null mice.
n3:mentions
n2:19088439
Subject Item
_:vb6461042
rdf:type
n3:Context
rdf:value
A previous report (>>7<<) has shown that resveratrol treatment leads to deacetylation of PGC-1α, suggesting that the catalytic activity of Sirt1 is increased by resveratrol.
n3:mentions
n2:17112576
Subject Item
_:vb6461043
rdf:type
n3:Context
rdf:value
Since AMPK activity increases the NAD-to-NADH ratio and Sirt1 activity (>>45<<), we measured the NAD-to-NADH ratio in C2C12 myotubes treated with resveratrol for 6 and 16 h.
n3:mentions
n2:18477450
Subject Item
_:vb6461044
rdf:type
n3:Context
rdf:value
PGC-1α is a coactivator for its own transcription (>>46<<), and, as a result, deacetylation of PGC-1α by Sirt1 should increase its ability to positively regulate PGC-1α transcription.
n3:mentions
n2:12764228
Subject Item
_:vb6461045
rdf:type
n3:Context
rdf:value
A previous report (>>7<<) showed that resveratrol did not induce PGC-1α transcription in Sirt1−/− mefs and concluded that Sirt1 is essential for resveratrol-induced transcription of PGC-1α.
n3:mentions
n2:17112576
Subject Item
_:vb6461046
rdf:type
n3:Context
rdf:value
Consistent with previous reports (>>47<<), −Sirt1 mefs had higher AMPK activity than +Sirt1 mefs (Fig.
n3:mentions
n2:18184747
Subject Item
_:vb6461047
rdf:type
n7:Section
dc:title
discussion
n7:contains
_:vb6461048 _:vb6461049 _:vb6461050 _:vb6461051 _:vb6461052 _:vb6461053 _:vb6461054 _:vb6461055 _:vb6461072 _:vb6461064 _:vb6461065 _:vb6461066 _:vb6461067 _:vb6461068 _:vb6461069 _:vb6461070 _:vb6461071 _:vb6461056 _:vb6461057 _:vb6461058 _:vb6461059 _:vb6461060 _:vb6461061 _:vb6461062 _:vb6461063
Subject Item
_:vb6461048
rdf:type
n3:Context
rdf:value
One target for treating metabolic disorders is Sirt1 (>>8<<), an NAD+-dependent protein deacetylase (48).
n3:mentions
n2:17167475
Subject Item
_:vb6461049
rdf:type
n3:Context
rdf:value
One target for treating metabolic disorders is Sirt1 (8), an NAD+-dependent protein deacetylase (>>48<<). Resveratrol, which was discovered in a small-molecule screen to be a Sirt1 activator (1), has drawn a great deal of interest for its therapeutic potential in treating metabolic disorders such as type 2 diabetes. By increasing
n3:mentions
n2:10693811
Subject Item
_:vb6461050
rdf:type
n3:Context
rdf:value
Resveratrol, which was discovered in a small-molecule screen to be a Sirt1 activator (>>1<<), has drawn a great deal of interest for its therapeutic potential in treating metabolic disorders such as type 2 diabetes.
n3:mentions
n2:12939617
Subject Item
_:vb6461051
rdf:type
n3:Context
rdf:value
In Sirt1 knockin mice (>>36<<), the Sirt1 transgene, which was expressed from the β-actin locus, is overexpressed in fat but not in muscle or liver, key organs affected in metabolic disorders.
n3:mentions
n2:17877786
Subject Item
_:vb6461052
rdf:type
n3:Context
rdf:value
More recently, transgenic mice in which the Sirt1 transgene is expressed from its own promoter have been reported by two independent groups (>>35<<,37). These transgenic mice, which have a more physiological expression pattern of the Sirt1 transgene than the knockin mice (36), are more insulin sensitive partly due to increased adiponectin levels and decreased hepatic steatosis.
n3:mentions
n2:18840364
Subject Item
_:vb6461053
rdf:type
n3:Context
rdf:value
More recently, transgenic mice in which the Sirt1 transgene is expressed from its own promoter have been reported by two independent groups (35,>>37<<). These transgenic mice, which have a more physiological expression pattern of the Sirt1 transgene than the knockin mice (36), are more insulin sensitive partly due to increased adiponectin levels and decreased hepatic steatosis.
n3:mentions
n2:18599449
Subject Item
_:vb6461054
rdf:type
n3:Context
rdf:value
These transgenic mice, which have a more physiological expression pattern of the Sirt1 transgene than the knockin mice (>>36<<), are more insulin sensitive partly due to increased adiponectin levels and decreased hepatic steatosis.
n3:mentions
n2:17877786
Subject Item
_:vb6461055
rdf:type
n3:Context
rdf:value
Surprisingly, the transgenic mice developed by Banks et al. (>>35<<), which are congenic in C57BL6/J background, have reduced metabolic rate and body temperature and have the same amount of fat despite eating less.
n3:mentions
n2:18840364
Subject Item
_:vb6461056
rdf:type
n3:Context
rdf:value
Nevertheless, this study shows that the function of Sirt1 on energy balance may be opposite of what was previously thought (>>7<<) and would be predicted if the central target of resveratrol was Sirt1.
n3:mentions
n2:17112576
Subject Item
_:vb6461057
rdf:type
n3:Context
rdf:value
One possible explanation for the reduced fat mass in the knockin mice (>>36<<) is that expression of the Sirt1 transgene from the β-actin locus impaired adipocyte differentiation during development (49).
n3:mentions
n2:17877786
Subject Item
_:vb6461058
rdf:type
n3:Context
rdf:value
One possible explanation for the reduced fat mass in the knockin mice (36) is that expression of the Sirt1 transgene from the β-actin locus impaired adipocyte differentiation during development (>>49<<). Therefore, the Sirt1 knockin model (36) cannot distinguish between a direct effect of Sirt1 and an indirect one caused by reduced fat.
n3:mentions
n2:15175761
Subject Item
_:vb6461059
rdf:type
n3:Context
rdf:value
Therefore, the Sirt1 knockin model (>>36<<) cannot distinguish between a direct effect of Sirt1 and an indirect one caused by reduced fat.
n3:mentions
n2:17877786
Subject Item
_:vb6461060
rdf:type
n3:Context
rdf:value
Resveratrol has been reported to affect the activities of many enzymes (>>11<<) including AMPK (12). To evaluate the possibility that the effects of resveratrol are mediated by AMPK, we studied the effects of resveratrol in mice deficient in AMPKα1 or -α2.
n3:mentions
n2:18421779
Subject Item
_:vb6461061
rdf:type
n3:Context
rdf:value
Resveratrol has been reported to affect the activities of many enzymes (11) including AMPK (>>12<<). To evaluate the possibility that the effects of resveratrol are mediated by AMPK, we studied the effects of resveratrol in mice deficient in AMPKα1 or -α2.
n3:mentions
n2:17438283
Subject Item
_:vb6461062
rdf:type
n3:Context
rdf:value
The ability of AMPK to increase NAD and the NAD-to-NADH ratio (>>45<<) may also explain how resveratrol treatment can lead to Sirt1 activation without directly activating it.
n3:mentions
n2:18477450
Subject Item
_:vb6461063
rdf:type
n3:Context
rdf:value
Moreover, AMPK can activate PGC-1α, one of Sirt1 substrates, by directly phosphorylating it (>>50<<,51), indicating that activation of AMPK can affect the Sirt1-dependent pathways in multiple ways.
n3:mentions
n2:19262508
Subject Item
_:vb6461064
rdf:type
n3:Context
rdf:value
Moreover, AMPK can activate PGC-1α, one of Sirt1 substrates, by directly phosphorylating it (50,>>51<<), indicating that activation of AMPK can affect the Sirt1-dependent pathways in multiple ways.
n3:mentions
n2:12181572
Subject Item
_:vb6461065
rdf:type
n3:Context
rdf:value
Whether it is the direct target of resveratrol or not, it appears that Sirt1 can also function upstream of AMPK in HepG2 hepatocytes and HEK293T cell line (>>26<<,27). Since Sirt1 is not required for resveratrol-mediated activation of AMPK activation in mefs (Fig. 6A), it is possible that Sirt1 is upstream of AMPK only in certain cell types.
n3:mentions
n2:18482975
Subject Item
_:vb6461066
rdf:type
n3:Context
rdf:value
Whether it is the direct target of resveratrol or not, it appears that Sirt1 can also function upstream of AMPK in HepG2 hepatocytes and HEK293T cell line (26,>>27<<). Since Sirt1 is not required for resveratrol-mediated activation of AMPK activation in mefs (Fig. 6A), it is possible that Sirt1 is upstream of AMPK only in certain cell types.
n3:mentions
n2:18687677
Subject Item
_:vb6461067
rdf:type
n3:Context
rdf:value
Resveratrol-induced physical endurance has been largely attributed to increased mitochondrial function (>>7<<). Certainly, converting muscle fiber to mitochondria-rich, slow-twitch fibers increases physical endurance (52).
n3:mentions
n2:17112576
Subject Item
_:vb6461068
rdf:type
n3:Context
rdf:value
Certainly, converting muscle fiber to mitochondria-rich, slow-twitch fibers increases physical endurance (>>52<<). However, it is also likely that the glycogen content in skeletal muscle, which is known to be increased by AMPK activity (34), also plays a role in resveratrol-induced physical endurance.
n3:mentions
n2:16847346
Subject Item
_:vb6461069
rdf:type
n3:Context
rdf:value
However, it is also likely that the glycogen content in skeletal muscle, which is known to be increased by AMPK activity (>>34<<), also plays a role in resveratrol-induced physical endurance.
n3:mentions
n2:9529258
Subject Item
_:vb6461070
rdf:type
n3:Context
rdf:value
We do not have a clear explanation for this, but one possibility is that the antiadipogenic function of Sirt1 (>>49<<) is being driven by the resveratrol-AMPKα1-Sirt1 pathway in AMPKα2−/− mice.
n3:mentions
n2:15175761
Subject Item
_:vb6461071
rdf:type
n3:Context
rdf:value
In skeletal muscle, the AMPKα1 isoform makes up only ∼20% of the total AMPK activity (>>17<<), and, yet, resveratrol-induced mitochondrial biogenesis (Fig.
n3:mentions
n2:10698692
Subject Item
_:vb6461072
rdf:type
n3:Context
rdf:value
One possible explanation is that in addition to the AMPKα2 activity, a crosstalk between the skeletal muscle and either the nervous system or fat, where the AMPKα1 isoform is predominant (>>17<<), is required for resveratrol-induced mitochondrial biogenesis or reduction of ROS in skeletal muscle.
n3:mentions
n2:10698692
Subject Item
_:vb6461073
rdf:type
n7:Section
dc:title
research design and methods
n7:contains
_:vb6461074 _:vb6461075 _:vb6461076 _:vb6461077 _:vb6461078 _:vb6461079 _:vb6461080
Subject Item
_:vb6461074
rdf:type
n3:Context
rdf:value
AMPKα2−/− (>>22<<) and AMPKα1−/− (28) mice were backcrossed to C57BL/6J for at least six generations before this study.
n3:mentions
n2:12511592
Subject Item
_:vb6461075
rdf:type
n3:Context
rdf:value
AMPKα2−/− (22) and AMPKα1−/− (>>28<<) mice were backcrossed to C57BL/6J for at least six generations before this study.
n3:mentions
n2:14573616
Subject Item
_:vb6461076
rdf:type
n3:Context
rdf:value
and fed a high-fat diet (40% calories from fat; Bio-serv) or a high-fat diet supplemented with resveratrol (400 mg · kg−1 · day−1; Orchid Chemicals and Pharmaceuticals) for 12 weeks as previously described (>>7<<). All experiments were approved by the National Heart, Lung, and Blood Institute Animal Care and Use Committee.
n3:mentions
n2:17112576
Subject Item
_:vb6461077
rdf:type
n3:Context
rdf:value
Reactive oxygen species (ROS) levels were determined in muscle extracts using the ROS-sensitive fluorescent dye dichlorodihydrofluorescein (DCF), as previously described (>>29<<,30). Briefly, oxidation-insensitive dye (carboxy-DCFDA) was used as a control to ensure that changes in the fluorescence seen with the oxidation-sensitive dye (H2DCFDA) were due to changes in ROS production.
n3:mentions
n2:17609368
Subject Item
_:vb6461078
rdf:type
n3:Context
rdf:value
Reactive oxygen species (ROS) levels were determined in muscle extracts using the ROS-sensitive fluorescent dye dichlorodihydrofluorescein (DCF), as previously described (29,>>30<<). Briefly, oxidation-insensitive dye (carboxy-DCFDA) was used as a control to ensure that changes in the fluorescence seen with the oxidation-sensitive dye (H2DCFDA) were due to changes in ROS production.
n3:mentions
n2:14766800
Subject Item
_:vb6461079
rdf:type
n3:Context
rdf:value
Sirt1−/− mouse embryonic fibroblasts (mefs) (>>31<<) were maintained in DMEM supplemented with 10% fetal bovine serum.
n3:mentions
n2:12482959
Subject Item
_:vb6461080
rdf:type
n3:Context
rdf:value
AMPKα1/α2 double knockout and wild-type mefs were derived as previously described (>>32<<). Cells were treated with 50 μmol/l resveratrol (Sigma) or DMSO for the indicated time. C2C12 myoblast cells (ATCC) were maintained in DMEM and 10% FBS. To generate C2C12 myotubes, a confluent culture of C2C12 cells were grown in DMEM and
n3:mentions
n2:16809770
Subject Item
_:vb419250061
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
53
n3:hasRelevantPaperId
n2:17086191
Subject Item
_:vb419250062
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
49
n3:hasRelevantPaperId
n2:17112576
Subject Item
_:vb419250063
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
37
n3:hasRelevantPaperId
n2:12939617
Subject Item
_:vb419250064
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
29
n3:hasRelevantPaperId
n2:22055504
Subject Item
_:vb419250065
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
26
n3:hasRelevantPaperId
n2:22560220
Subject Item
_:vb419250066
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
21
n3:hasRelevantPaperId
n2:22304913
Subject Item
_:vb419250067
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
19
n3:hasRelevantPaperId
n2:20061378
Subject Item
_:vb419250068
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
18
n3:hasRelevantPaperId
n2:19262508
Subject Item
_:vb419250069
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
16
n3:hasRelevantPaperId
n2:17438283
Subject Item
_:vb419250070
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
16
n3:hasRelevantPaperId
n2:20519126
Subject Item
_:vb419250071
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
15
n3:hasRelevantPaperId
n2:15744310
Subject Item
_:vb419250072
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
14
n3:hasRelevantPaperId
n2:20197054
Subject Item
_:vb419250073
rdf:type
n3:RelevantBibliographicResource
n3:RelevantScore
14
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