Abstracts
Oral – [A-10-476-1]
Role of glutathione system in biochemical and metabolic
functions of hepatocytes-like cells differentiated from
human bone marrow mesenchymal stem cells
Abdolamir Allameha, Ahmadi-Ashtiani Hamidrezaa,
Kazemnejad Somaieha, Soleimani Masouda, Gharebaghian Ahmadb,
Rastegar Hosseinc, Ismaeili Shahnazb, Hedayati Mehdib
a
POB 14115–111, Tehran, Iran
b
Tehran, Iran
c
Ministry of Health, Tehran
E-mail address: allameha@modares.ac.ir (A. Allameh)
Introduction: Glutathione plays a major role in various cellular
functions. However our information about glutathione system involvement in biochemical and metabolic functions of hepatocyte-like cells
during differentiation from mesenchymal stem cells is scanty. This study
is focused on the role of glutathione on biological activity of hepatocytes
during differentiation.
Methods: Differentiation of MSCs into hepatocytes was induced using
standard protocol in a specific culture media. The non-differentiated
and differentiated cells were collected at different time intervals (0, 2, 7
and 14 days) and the rate of cell viability (Trypan blue test) and
proliferation (Brdu assay) were monitored. After characterization of
hepatocytes and their progenitor MSCs, the levels of cellular albumin,
alpha-fetoprotein (AFP), cytochrome P450 (CYP), reduced glutathione
(GSH), oxidized glutathione (GSSG), total glutathione, GSH/GSSG ratio,
GSH peroxidase (GPX) and GSH reductase (GR) were measured and
compared to respective controls. Albumin and urea and secretion into the
culture media were monitored as the biosynthesis capacity of the
hepatocytes.
Results: The growth curve of the differentiated hepatocytes revealed
that the MSCs were successfully differentiated and the proliferation
rate was normal during three-week culture. Expression of liver function
factors such as; albumin, AFP and CYP were increasing following
hepatogenic differentiation of the MSCs. It was also found that
glutathione modifiers can influence cell function and metabolism.
Excretion of urea and albumin into the culture media as index of
metabolic activity was evident on day 7 of differentiation. Treatment of
the cells with glutathione modifiers showed that the cellular GSH play a
major role in cell survival and biological activity.
Conclusions: Cellular glutathione as a major antioxidant factor could
play a major role in viability, longevity and metabolic function of
hepatocytes differentiation from adult stem cells.
Keywords: Biochemical, Metabolic, Hepatocytes, Stem cell,
Differentiation
doi:10.1016/j.clinbiochem.2011.08.1061
Oral – [A-10-1458-2]
Promoting chemical safety through toxicology education in
the classroom
Rahmat Awanga, Nida Besbellib, Agneta Sunden Bylehnc,
Birger Heinzowd, Lillian Corrae
a
National Poison Centre, Universiti Sains Malaysia, Malaysia
b
United Nations Environment Programme, Geneva
c
LGA-SH, State Agency for Health of Land Schleswig-Holstei, Germany
d
New Zealand Poison Centre, New Zealand
e
International Society of Doctors for the Environment, Argentina
E-mail address: rahmatawang@yahoo.com (R. Awang)
In many developing countries, infants and children are particularly vulnerable to pesticides and other toxic chemicals because their
bodies are smaller and still developing. Children also face greater
S43
exposures than adults due to their hand-to-mouth behaviors.
Children living in farming areas or whose parents work in agriculture
suffer greater pesticides exposure than other children. Non-chemical
methods exist to prevent or manage pests. Integrated Pest Management (IPM) and Integrated Vector Management (IVM) apply different
methods, trying to reduce the use of chemicals to a minimum.
Despite this, pesticides are much used in farming economies today,
and for managing vectors of disease, especially in developing
countries and countries with economies in transition. Pesticides are
applied to kill pests, but they can also adversely affect non-target
organisms, including humans.
In the developing world most people live in rural areas and
children's exposure to pesticides is unavoidable since farms are
homes as well as workplaces. Educational materials that is aimed
specifically at creating awareness among young students about
potential side effects of chemicals and help reduce careless use and
handling of pesticides and other chemicals are not available.
This has brought the attention of the United Nations Environmental Programme (UNEP) and the World Health Organisation
Regional Office for Europe (WHO Euro) and the Malaysian National
Poison Centre of Universiti Sains Malaysia (MNPC) with initial
support from the International Union of Pure and Applied Chemistry
(IUPAC) to develop an innovative teaching courseware utilizing
multimedia that will help provide basic understanding of toxicology
and awareness of the need for protective and precautionary
measures to minimise adverse effects on human health and the
environment.
This paper highlights how young children learn toxicology and
interact with the courseware. The experience gained through its
testing in Malaysia and Argentina will also be described.
doi:10.1016/j.clinbiochem.2011.08.1062
Oral – [A-10-818-1]
Effects of pesticide on the thyroid hormones of pesticide sprayers
living in Mazandaran
Fatemeh Tohidia, Farzaneh Farrokhib, Ali Taravatic
a
Department of Modern Sciences and Technologies Mashhad University
of Medical Science Mashhad, Iran
b
Islamic Azad University, Sari Branch, Sari, Iran
c
Islamic Azad University, Jouybar Branch, Jouybar, Iran
E-mail addresses: tohidiF891@mums.ac.ir (F. Tohidi),
ali_taravatie@yahoo.com (A. Taravati)
Introduction: Pesticides are occasionally used indiscriminately in
large amounts causing environmental pollution. Pesticides cause
adverse effects on different body systems, including hematological
and endocrine systems. The ability of organophosphate pesticides to
disturb thyroid gland function has been demonstrated by experimental studies on animal, but evidence of such effects on human
remains scarce. The purpose of this study was to investigate the
association between exposure to pesticide and serum levels of
thyroid hormones in pesticide sprayers.
Methods: Blood samples were obtained from 40 cases and 20
controls after overnight fasting. Glucose, urea, acid uric and lipid
profile were measured by enzymatic methods. TSH, T3 and T4 levels
were determined by immunoassay kits.
Results: The TSH level in sprayers (2.64 ± 1.50 μM, P = 0.007) was
significantly increased compared with the control subjects (1.33 ±
0.83). T3 and T4 levels of sprayers were lower than controls.
Conclusion: These results suggest that exposure to organophosphate pesticides may be responsible for increasing TSH level and
decreasing T3 and T4 serum hormone levels, therefore supporting the
hypothesis that organophosphate pesticides cause hypothyroidism in
S44
Abstracts
pesticide sprayers. So administration of supplementary nutrients or
improving defense system in these subjects is advised.
Keywords: Hypothyroidism, Pesticide, TSH, Immunoassay
doi:10.1016/j.clinbiochem.2011.08.1063
Oral – [A-10-831-1]
Protective role of N-acetyl-cysteine on diazinon induced oxidative
stress in rat kidney
Farideh Izadia, Mahvash Jafarib, Alireza Asgarib, Maryam Salehib
a
Department of Biology, Islamic Azad University, Tehran, Iran
b
Baqiatall University, Iran
E-mail addresses: Izadi_FArideh@yahoo.com (F. Izadi),
jafari@bmsu.ac.ir (M. Jafari)
Introduction: Organophosphate pesticides (OP) may cause generation of reactive oxygen species (ROS), which may lead to oxidative stress.
Diazinon (DZN) is an organophosphorus insecticide that has been
widely used throughout the world with applications in agriculture and
horticulture for controlling insects. N-acetyl-l-cysteine (NAC), precursor
of reduced glutathione (GSH), is an antioxidant and free-radical
scavenger. In this study, the possible protective role of NAC in DZNinduced toxicity was evaluated in male Wistar rats.
Materials and methods: In vivo experimental groups were: control
group (corn oil as diazinon solvent), DZN group (100 mg/kg, i.p.), NAC
(160 mg/kg, i.p.) group and NAC–DZN group. 24 hours after injection,
animal to ether anesthesia and kidney removed and superoxide
dismutase (SOD) and catalase (CAT) activities, as well as GSH and
malondialdehyde (MDA) levels were determined by biochemical
methods.
Results: The result showed that SOD and CAT activities and MDA level
were increased, while GSH level was decreased in DZN treated rats as
compared to control. NAC–DZN groups were found to improve these
disorders.
Conclusion: The results suggest that NAC provides protection against
DZN-induced oxidative stress and prevents lipid peroxidation in kidney.
Keywords: Diazinon, N-acetyl-cysteine, Antioxidant system, Rat,
Kidney
doi:10.1016/j.clinbiochem.2011.08.1064
Oral – [A-10-1001-1]
Activation of Nuclear Factor-kappaB by doxorubicin leads to
inhibition of extrinsic pathway of apoptosis in H9c2 cells
Leila Hosseinzadeh, Gholamreza Karimi,
Fatemeh Mosaffa, Mitra Rezaee Asgharian
Faculty of Pharmacy, Kermanshah University of Medical Sciences,
Kermanshah, Iran
E-mail addresses: L_hoseinzadeh@yahoo.com (L. Hosseinzadeh),
gkarimi@mums.ac.ir (G. Karimi), ft_mosaffa@yahoo.com (F. Mosaffa)
Introduction: Doxorubicin (DOX), a chemotherapeutic agent, can
give rise to serve cardiotoxicity by inducing apoptosis. The exact
mechanism of DOX-induced apoptosis has not been fully understood
yet. Here, we evaluated the effect of DOX on some important genes
involved in apoptosis and also Nuclear Factor-kappaB (NF-κB) activity in
H9c2 cardiomyoblast cells.
Materials and methods: Cell viability was determined by MTT.
Apoptosis was assessed by annexine V/PI double staining. Real time RT-
PCR was used to evaluate the expression of genes. The DNA-binding
capacity of NF-κB was examined by ELISA.
Results: DOX-mediated cytotoxiocity is executed by inducing
apoptotic cell death. DOX induced apoptosis by down regulating antiapoptotic Bcl-2 and up-regulating pro-apoptotic Bax. Consequently, the
ratio of Bax to Bcl-2 is significantly increased upon treatment with DOX
leading to increase caspse-9, the initiator caspase of mitochondrial
pathway for apoptosis, expression. However, it had no effect on the
expression of caspase-8 the mediator of extrinsic pathway. DOX wasn't
able to inhibit expression of anti-apoptotic gen cIAP1. Also, we observed
that NF-κB activity was sharply increased by incubation in the presence of
DOX. Previous studies have shown that depending on the cell models,
apoptosis from DOX can follow different pathways. Our findings
demonstrated that apoptosis induced by doxorubicin occurred through
intrinsic pathway rather than by extrinsic pathway. It was indicated that
NF-κB induces multiple factors to regulate apoptosis including cIAP1.
Conclusion: Based on these results, it is possible that NF-κB activation
suppresses extrinsic pathway of apoptosis through blocking the cIAP1
down-regulation and consequently caspase-8 activation by DOX in H9c2
cells.
Keywords: Doxorubicin, NF-kappaB, Apoptosis, H9c2 cells
doi:10.1016/j.clinbiochem.2011.08.1065
Oral – [A-10-1026-2]
Apoptosis induction of methysulfonylmethane in human
gastrointestinal cancer cell lines
Naser Jafari, Shahab Bohlooli
Ardabil University of Medical Sciences, Ardabil, Iran
E-mail address: n.jafari@arums.ac.ir (N. Jafari)
Introduction: Methylsulfonylmethane (MSM) is an organic sulfur
containing compound that occurs naturally in a variety of fruits,
vegetables and unpasteurized milk. It has been used as a supplement
in the last two decades. Recently it has been suggested that MSM has
clinical potential as a non-toxic agent effective against metastatic
melanoma. Considering the preventive effects of this substance on
tumor onset and nontoxic to healthy body we have investigated in vitro
effects of methylsulfonylmethane on apoptosis induction in human
gastrointestinal cancer cell lines.
Methods: The human cancer cell lines including, AGS, HepG2 and
KYSE30 were cultured and incubated until confluence. The cells were
removed and seeded in 96-well plates at a density of 1 × 103 cells/well
and incubated overnight, and then treated with (21–30) mg/ml MSM. To
stain apoptotic cells, the plates were centrifuged. The EB/AO dye mix was
prepared in PBS and 10 μl was added to each well and cells were viewed
and counted under an inverted fluorescence microscope.
Results: The results showed that after incubation at (21–29) mg/ml
of MSM for 24 h the cells displayed a series of morphological changes
including condensation and fragmentation of chromatin and nucleus,
and formation of apoptotic bodies which were designated as typical
evidence of apoptotic bodies. In contrast control cells exhibited a normal
appearance. There was a significant increase of apoptotic cells in cancer
cells after treating with MSM.
Conclusion: The results demonstrated that MSM exerts its anticancer and cytotoxic effect by inducing apoptotic cell death.
Keywords: Apoptosis, Methylsulfonylmethane, Cancer
doi:10.1016/j.clinbiochem.2011.08.1066