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HISTOPATHOLOGICAL STUDY OF NITRATE ION
EFFECT ON PANCREAS EXPERIMENTALLY IN LABORATORY
MICE.
Salma S.AL- Anni*, Zainab R. Zghair**, Mohanad D. Al- jaboore***, Eman
K. Khalel***
*Ministry of science and Technology, Environment and Water office ,Baghdad, Iraq
** Zoonosis Unit ,College of Veterinary Medicine,university of Baghdad,Baghdad,Iraq
***department of Public health College of Veterinary medicine,University of Falluja,Iraq.
(Received 4 May 2016 ,Accepted 25July 2016)
Key words :- Nitrate , Histopathological change,Diabetes
ABSTRACT
This study was designed to explore the effect of nitrate ion on induction of diabetes
experimentally in laboratory mice.Fifteen white mice, 7-8 week olds were randomly divided
into three equal groups. First group was administrated orally o.3 ml of nitrateion
(concentration was 0.25gm/10ml D.W) daily for one month. Second group was administrated
orally o.3 ml of potassium nitrate (concentration was 0. 5gm/10ml D.W) daily forone
month. Third group was administrated orally with 0.3 ml of distilled water (D.W) daily for
one month.After 30 post administrationthe concentration of blood glucose of infected
animals treated with nitrate ion increase in both first and second groups when compared with
control group.
The histopathological change study showed pathological lesions in pancreasof first group
treated with nitrate ion showed the exocrine region and islets of Langerhans with damaged
due to necrosis, while the second group wasinfected with nitrate ion showed the exocrine
region and islets of Langerhans with necrosis and congestion of blood vessel.
INTRODUCTION
Diabetes mellitus is one of the most common endocrine diseases, associated
with a group of metabolic disorder characterized by chronic hyperglycemia with
disturbances of carbohydrate, lipids, and protein metabolism resulting from defects in
insulin secretion, insulin action, or both (1). Type 1 diabetes is thought to be inherited
in genetically susceptible individuals by environmental factors such as viral, toxic or
chemical agents that lead to autoimmune destruction of B-cells, resulting in the formation
of altered protein components. This material is a foreign antigen to the immuno system,
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establishing the basis for an autoimmune reaction against the cell of origin the (B-cell)
(2),(3). Nitrate ion Occurrence in food, food supplements and medicines Potassium nitrate
and nitrite are used as food preservatives, mainly for curing meats. (4) investigated the
toxicity of potassium nitrite (KNO2) andconcluded that the sub-chronic toxicity.
Nitrate ion content is an important quality characteristic of vegetables. Vegetable nitrate
content is of interest to governments and regulators owing to the possible implications for
health and to check that controls on the content are effective. Nitrate itself is relatively nontoxic
but its metabolites may produce a number of health effects. Nitrate was perceived as a
purely harmful dietary component which causes infantile methaemoglobinaemia,
carcinogenesis and possibly even teratogenesis (5). Other complicationschildren who are
exposed to large amounts of nitrate or nitrite might be at an increased risk of developing
childhood diabetes, experience recurrent diarrhea or contract upper respiratory infections.
Nitrates are also dangerous for unborn children. Mothers with diets rich in nitrite and nitrate
are more likely to have babies with slow intrauterine growth, heart defects and Sudden Infant
Deaths Syndrome (SIDS) (6).
MATERIALS AND METHODS
1- Nitrate ion preparation:
-Nitrate ion dissolved in distilled water (D.W) to prepare doses one of them 0.25 gram/10
ml. of D.W and the other dose 0.5 g/10 ml.10 cc of D.W .
- Experimental Design:
Fifteen white mice both sexes, 7-8 week olds were randomly divided into three groups
equally and treated as follows:
1- First group was administrated orally o.3 ml of nitrateion (concentration was
0.25gm/10ml) daily for one month.
2- Second group was administrated orally o.3 ml of nitrate ion (concentration was 0.
5gm/10ml) daily for one month.
3- Third group was administrated orally with 0.3 ml of distilled water (D.W) daily for one
month.
At day 30 post administratation all animals were sacrificed and collection of blood from each
animal 0.5-1milliliter of blood to get serum that using in blood sugar measuring , and pieces
from pancreas were taken for fixed in 10% neutral buffer saline 72 hours for
histopathological changes examination according to(7).
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RESULTS
1-Measurements of blood sugar treated animals:
The concentrations of blood serum sugar treated animals with potassium nitrate 0.25 g/10 ml
and 0.5 g/10 ml with control group as showed in table (1) increase in the concentrations of
blood sugar for both first and second groups when compared with control group.
No. Infected with0.25 g/10 ml
of potassium nitrate
Infected with0.5 g/10
ml of potassium nitrate
control
1 127 141 118
2 124 138 116
3 154 149 105
4 173 153 112
5 195 171 115
Total 154.6 150.4 113.2
Table 1: show the titer of serum blood glucos infected animals with potassium nitrate 0.25
g/10 ml and 0.5 g/10 ml with control group
2- Histopathological change study:
- Pancreas-Showed that exocrine region and islets ofLangerhans with damaged β cells due
to necrosisof the first group (fig: 1), and the second group appeared the exocrine region and
islets of Langerhans with damaged β cells due to necrosis and a decreased number of β cells
and congestion of blood vessel(fig: 2).
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Fig1:Histopathological section in pancreases of one animal infected
with potassium nitrate 0.25 g/10 ml showed the exocrine region and
islets of Langerhans with damaged β cells due to necrosis and a
decreased number of β cells ( ) (H&EX400).
Fig2:Histopathological section in pancreases of one animal infected
with potassium nitrate 0.5 g/10 ml showed the exocrine region and
islets of Langerhans with damaged β cells due to necrosis and a
decreased number of β cells and congestion of blood vessel ( )
(H&EX400).
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DISCUSSION
From our data, we concluded, the increased blood sugar is good indicators for the
diagnostic of tissue injury of pancreas by the toxicity of potassium nitrates .The sub-chronic
toxicity of potassium nitrite has been investigated by (4) study, 6 week old Wistar rats were
treated with potassium nitrite (concentrations not specified) in their drinking water for 90
days.There were nodeaths but hypertrophy of the was found in all treatedgroups (increased
incidence and severity with increasing levels of nitrite in the water (8). A number of studies
evaluated possible associations between developmental end points and exposure to
nitrate.The results provide some evidence of nitrate-related developmental effectspossible
confounding by other potential toxicants was not evaluated, and most studies did not account
for dietary nitrate or nitrite intake, which is typically the major source of ingested nitrate and
nitrite. Some studies reported significant associations between selected developmental end
points and nitrate in drinking water sources. Other studies found no evidence of associations
between nitrate and risk of developmental effects. Finally there are no studies about
induction of nitrate ion to diabetes. There is deficiency in references about this study.
دراسة إمراضیھ نسجیة لتأثیر ایون النترات على عضو البنکریاس تجریبیا فی الفئران المختبریة
سلمى سھام العانی*، زینب رزاق زاھر**، مھند ضیاء الجبوری*** ، ایمان خلف خلیل***
*وزارة العلوم والتکنلوجیا ، دائرة البیئة والمیاه .بغداد ،العراق
**وحدة الامراض المشترکة ، کلیة الطب البیطری ،جامعة بغداد ،بغداد ،العراق
*** قسم الصحة العامة ، کلیة الطب البیطری، جامعة الفلوجة،العراق .
الخلاصة
صممت ھذه الدراسة للتحری عن تاثیر ایون النترات فی احداث مرض السکری تجریبیا فی الفئران المختبریة، استخدم
7 اسابیع عشوائیا الى ثلاثة مجامیع متساویة. اعطیت - فی الدراسة خمسة عشر فارا ابیض من کلا الجنسین وبعمر 8
المجموعة الاولى 0.3 مللترمن نترات البوتاسیوم عن طریق الفم (وبترکیز 0.25 ملغرام/ 10 مللتر من الماء المقطر). اما
0 ملغرام/ 10 مللتر من الماء . المجموعة الثانیة فقداعطیت 0.3 مللترمن نترات البوتاسیوم عن طریق الفم (وبترکیز 5
المقطر)، والمجموعة الثالثة فقداعطیت 0.3 مللترمن الماء المقطر عن طریق الفم یومیا ولمدة شھر. وبعد ثلاثون یوما
لوحظ ارتفاع ارتفعت معدلات سکر الدم للحیوانات المعاملة بایون النترات للمجموعتین الاولى والثانیة مقارنة بمجموعة
السیطرة.
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اظھرت الدراسة المرضیة النسجیة وجود افات مرضیة فی البنکریاس، واظھرتالمجموعة الاولى تحطم المنطقةخارجیة
الافرازنتیجة التنخروتناقص اعداد خلایا بیتا ، بینما اظھرت المجموعة الثانیة تحطم المنطقة خارجیة الافرازوجزیرات
لانکرھانز مع التنخرواحتقان الاوعیة الدمویة.
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