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Pt NPs Diabetes

Pt NPs Diabetes

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Published by Webb Norfleet

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Published by: Webb Norfleet on Apr 23, 2013
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MEETING ABSTRACT Open Access
Anti-diabetes effect of water containinghydrogen molecule and Pt nanoparticles
Sanetaka Shirahata
1,2*
, Takeki Hamasaki
1
, Keisuke Haramaki
2
, Takuro Nakamura
1
, Masumi Abe
1
, Hanxu Yan
2
, Tomoya Kinjo
2
, Noboru Nakamichi
3
, Shigeru Kabayama
3
, Kiichiro Teruya
1,2
From
22nd European Society for Animal Cell Technology (ESACT) Meeting on Cell Based TechnologiesVienna, Austria. 15-18 May 2011
Background
Electrochemically reduced water (ERW) contains a lot of hydrogen molecule (H
2
) and scavenges reactive oxygenspecies (ROS) to protect DNA from oxidative damage[1]. ERW also contains small amounts of Pt nanoparti-cles (NPs) and elongates the lifespan of 
C. elegans
[2].Pt NPs are newly recognized multi-functional ROS sca- vengers [3]. ERW exhibits anti-diabetes effects
in vitro
and
in vivo
[4-6][7]. We proposed mineral nanoparticle active hydrogen reduced water hypothesis to explain theactivation mechanism of H
2
to hydrogen atom (H)[4].Recently, H
2
has been reported to scavenge ROS andsuppress a variety of oxidative stress-related diseases [8],however, the action mechanism of H
2
has not been clar-ified thoroughly. Here, we examined anti-diabetes effectsof H
2
and Pt NPs.
Materials and methods
Pt NPs of 2-3 nm sizes were synthesized from H
2
PtCl
6
by the citrate reduction method. L6 rat myoblast cells(1.2 x 10
5
cells) were inoculated into a 35 mm culturedish and a day later, the cells were treated with or with-out 25mM N-acetylcystein in the presence of BES-H2O2, a H
2
O
2
-specific detection reagent in DMEM for2 h. After washing the cells, molecular hydrogen treat-ment was performed in a dark condition by cultivatingcells in a fresh DMEM medium in a mixed gas incuba-tor under an atmosphere of 75%N
2
/20%O
2
/5%CO
2
or75%(H
2
and N
2
mixed gas)/20%O
2
/5%CO
2
for 1.5 h, fol-lowed by flowcytometric analysis. In this condition, cul-ture medium contained maximum 0.4-0.5 ppm of dissolved hydrogen. Glucose uptake of differentiatedmyotube L6 cells was examined after treating the cellswith
3
H-2-deoxyglucose for 10 min. Gene expression of catalase (CAT), glutathione peroxidase (GPx) andhemoxoigenase (HO-1) was examined using RT-PCRmethod. Three weeks old type 2 diabetes model mice(KK-
 A
 y 
) were fed H
2
and/or Pt Nps-containing water
ad lib
for 6 weeks.
Results
H
2
stimulated glucose uptake into L6 cells. Pt NPs cata-lyzed the activation of H
2
to hydrogen atom (H) to sca- venge DPPH radical
in vitro
. The combined use of molecular hydrogen and Pt NPs resulted in extremely stimulated glucose uptake into L6 cells, suggesting thatH produced from H
2
by catalyst action of Pt NPs regu-lated glucose uptake signal transduction. As oppose tothe paper by Ohsawa et al.[8], H
2
of 25 to 75% concen-tration in the mixed gas significantly scavenged intracel-lular H
2
O
2
in rat fibroblast L6 cells (Figure1) andinduced the gene expression of antioxidative enzymessuch as CAT, GPx and HO-1 via activation of Nrf2 (Fig-ure2). H
2
, Pt NPs and their combination significantly suppressed the levels of fasting blood glucose andimproved the impaired sugar tolerance abilities of obeseinsulin-resistant type 2 diabetic KK-
 A
 y 
mice.
Conclusion
H
2
, Pt NPs, and their combined use resulted in activa-tion of glucose uptake signal transduction pathways andstimulation of glucose uptake into L6 myotubes. In thegroups of H
2
, Pt NPs and their combined use groups,blood sugar levels and impaired sugar tolerance of type2 diabetes model mouse (KK-
 A
 y 
) were significantly improved, suggesting that H
2
, Pt NPs and H are redoxregulation factors in animal cells.
* Correspondence:sirahata@grt.kyushu-u.ac.jp
1
Department of Bioscience and Biotechnology, Faculty of Agriculture, KyushuUniversity, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, JapanFull list of author information is available at the end of the article
Shirahata
et al 
.
BMC Proceedings
2011,
5
(Suppl 8):P18http://www.biomedcentral.com/1753-6561/5/S8/P18
© 2011 Shirahata et al; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the CreativeCommons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, andreproduction in any medium, provided the original work is properly cited.
 
Figure 1
The scavenging effect of hydrogen molecule on intracellular hydrogen peroxide in rat myotube L6 cells. ***, p<0.001.
Figure 2
Induced gene expression of anitoxidative enzymes by hydrogen molecule. L6 myoblast cells were cultivated under an atmosphere of 75%N
2
or H
2
 /20%O
2
 /5%CO
2
for 2 h and gene expression was analyzed by RT-PCR. *, P<0.05
Shirahata
et al 
.
BMC Proceedings
2011,
5
(Suppl 8):P18http://www.biomedcentral.com/1753-6561/5/S8/P18Page 2 of 3

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