Annals of RSCB

Vol. XIV, Issue 1

RESEARCH ON API-DIET INFLUENCE OF LIPIDIC PROFILE IN

WISTAR RAT SUFFERING FROM HEPATOPATHY INDUCED BY
ACRYLAMIDE
C.V. Andrioiu1, V. Andrioiu2, Magdalena Cuciureanu3,
Rodica Cuciureanu4, Cristina Mihaela Ghiciuc1, Liliana Taru1,
Alina Manole5, Ctlina Elena Lupuoru1
1

DISCIPLINE OF PHARMACOLOGY-ALGESIOLOGY, FACULTY OF MEDICINE, GR. T. POPA

UNIVERSITY OF MEDICINE AND PHARMACY ROMANIA; 2 APIREGYA IMUNOSTIM PRIVATE
OFFICE, GORJ, ROMANIA; 3 DISCIPLINE OF PHARMACOLOGY-ALGESIOLOGY, FACULTY OF
DENTAL MEDICINE; 4DISCIPLINE OF ENVIRONMENT CHEMISTRY, HYGIENE AND NUTRITION,
FACULTY OF PHARMACY; 5DISCIPLINE OF PRIMARY HEALTH CARE AND EPIDEMIOLOGY,
FACULTY OF MEDICINE, GR. T. POPA UNIVERSITY OF MEDICINE AND PHARMACY ROMANIA

Summary
The aim of this experiment was to study the apidiet influence of hepatopathy induced by
acrylamide, in rats. The objectives were to assess the following parameters: cholesterol and
triglycerides, V-LDL, HDL, in terms of chronic hepatic injury induced by acrylamide, in rats. In
order to reduce factors that precipitate the progression of hepatic lesions, we administered
apitherapic products. Material and methods: Substances: apitherapic products - Apiregya,
ApiImunomod, ApiImunostim get from Stupina SRL (office and laboratories in the Balanesti
parish, Gorj county, Romania, phone 0253.270.221). Animals were manipulated under general
anesthesia with Thiopental. The levels of investigated parameters were measured using an automatic
analyzer (Aeroset, Abbott) and commercial kits (Abbott, USA). A number of 60 white rats Wistar
race was used, divided equally in six lots: the control lot standard nourishment (lot I), apidiet
control lot (lot II), apidiet and royal jelly (RJ) control lot (lot III), acryl amide lot (lot IV), apidiet
and acrylamide lot (lot V), acry amide, apidiet, and RJ lot (lot VI). After that, a number of 30
subjects were kept for study and research. Animals were kept in normal conditions for light, with
access to water and food. Toxic hepatopathy was experimentally induced by acryl amide
administered by intra-gastric gavage (hydrous solution, 50 mg/kg). Results and discussions: Apidiet
+ RJ administration (lot III) was efficient to keep the level of total cholesterol within normal limits.
Apidiet + RJ administration in rats with previous acryl amide intoxication was efficient to hold the
level of triglycerides within normal limits. No statistically significant differences between the
studied lots were registered. This fact demonstrated the efficacy of apidiet+RJ administration in lot
with previous acryl amide intra-gastric gavage, in order to maintain VLDL level. The increase of
HDL cholesterol level certified benefic effect of apidiet + RJ nourishment after a previous
intoxication with acryl amide. Conclusions: Authors recommend apitherapy with Apiregya,
ApiImunostim, and ApiImunomod in hepatic affections due to toxic nutrition, in order to normalize
the lipid profile.
Key words: apitherapy, pollen, hepatopathy, cholesterol, triglycerides.

dr_calin_andritoiu@yahoo.com
increased and various contain in essential
aminoacids, needful to life. Aminoacids are
(after A. Caillas, cited by L.A. Mrghita,
2005): arginine 5.70 %, histidine 2.4%,
isoleucine 4.50%, leucine 6.7%, lysine

Introduction
Pollen contains all of essential and
non-essential aminoacids with a role in
human nutrition (V. Andrioiu, 2005). High
biological value of pollen results from its
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Vol. XIV, Issue 1

5.7%, methionine 1.80%, phenylalanine

3.9%, threonine 4.0%, tryptophan 1.3%,
valine 5.7% (L.A. Mrghita, 2005).
Azotated substances were identified, also:
xanthine,
hypoxanthine,
geranine,
trimetylamine (L.A. Mrghita, 2005; V.
Andrioiu, 2005). Royal jelly (RJ) is a
resinous substance, with anti-mutagen,
hepato-protective, anti-inflammatory, antioxidant, and anesthetic effects. Apilarnil1 is
a new product from bees, with a
composition almost similar with RJ and
other bee products and represents a cocktail
of vitalizing and regenerative substances
used in therapy and needful for nutrition
and medical practice. It is obtained by
trituration of drone larvae collected in
special conditions, in day 7 of larvar life,
using a technology based on intensive
production, in order to turn to advantage in
natural state2 (N.V. Iliesiu, 19913).
Acrylamide was obtained for the first
time by Moureu in Germany, in 1893
(Eriksson, E., Acrylamide, 2005), and
industrial production was inaugurated by
American Cyanamid Inc., in 1954
(NICNAS, Acrylamide. Priority Existing
Chemical. Assessment Report No. 23,
2002). Acrylamide is used nowadays in the
drinking water and wastewater treatment,
plastic fabrication and some packs for food
products, paper and cellulose manufacture,
sugar refining, and raw oil processing
(Cuciureanu, R., Bulea, D., 2005, ***,
CERHR, 2005).

Material and methods

Necessary substances: apitherapic
products
(Apiregya,
ApiImunomod,
ApiImunostim), obtained from Stupina
SRL4; acrylamide, and thiopental. The
measurement of investigated parameters
level, and total serum proteins, respectively,
was performed using an automatic analyzer
(Aeroset, Abbott) and commercial kits
(Abbott, USA).
A number of 60 white rats, Wistar
race was used, divided equally in 6 lots (the
control lot standard nourishment (lot I),
apidiet control lot (lot II), apidiet and royal
jelly (RJ) control lot (lot III), acrylamide lot
(lot IV), apidiet and acrylamide lot (lot V),
acrylamide, apidiet, and RJ lot (lot VI).
Average weight: 250 g. Animals were kept
in normal conditions for light, with
unlimited access to water and food.
Toxic hepatopathy was experimentally induced by acrylamide, administered
by intra-gastric gavage (hydrous solution,
50 mg/kg). All the experiments performed
on laboratory animals (Wistar rats) were in
accordance with international ethical
settlements. Animals were anesthetized
using Thiopental, 50mg/kg, i.p., before the
biological samples collecting. In the end of
the study, animals were euthanatized after
general anesthesia and the death was
induced rapidly, without any agony
(Andrews E.J., AVMA Euthanasia, 1997).
Animals were sacrificed by anesthesia with
Thiopental in a dose of 1 ml/100g from
0.01% Thiopental solution. After the
Thiopental anesthesia, the thorax was
opened and blood was collected by heart
puncture, using a Vacuette collecting
system (R), in order to measure the
established parameters.
The aim of this study consisted in
experimental research on apidiet influence
in hepatopathy induced by acrylamide in
rats Wistar race. The objectives of the

The new product, active biologically, was named

apilarnil, suggesting its bee origin (api), larvar
provenance (lar), and researcher initials (nil).
The product was registered at State Patents and
Brands Office (O.S.I.M.), as trade mark, under this
name and no. 69389/1980.
2

Its finalization as an active biologic product,

collected by an intensive technology, and meant, as
raw material, for numerous utilizations in humans
and animals, represents a world performance, under
its form and achievement and utilization conditions.
3
Chapter 1: Contemporary apiculture turns to
diversification of bee production and its
capitalization. Subchapter 2: A bee potential with
profit perspectives: larvae, page 23.

The society office and laboratories are located in

Blaneti Parish, Gorj County, Romania, phone:
0253270221.

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Vol. XIV, Issue 1

experimental study were to assess the

parameters of protein electrophoresis, in
terms of chronic liver injury induced by
acrylamide. In order to reduce the factors
that accelerate the progression of hepatic
lesions, we administered apidiet and apidiet
with royal jelly.

Alpha-1 globulin. Co-administration

of acrylamide and apidiet (lot III)
determined the significant statistic decrease
of alpha-1 globulin, in comparison to the
control lot with standard food (lot I)
(28.640.22 versus 23.831.62, p<0.0001)
(Figure no. 3). Administration of
apidiet+RJ (lot II) determined a significant
statistic decrease of alpha-1 globulin,
comparatively to the control lot with
standard food (lot I) (28.640.22 versus
25.120.24, p<0.0001) (Figure no. 3). No
significant statistic differences for alpha-2
globulins level between the studied lots
were registered (Figure no. 4).
Beta-globulin. Co-administration of
acrylamide and apidiet (lot III) led to the
significant statistic decrease of betaglobulin, comparatively to the control lot
with standard food (lot I) (26.47 0.44
versus 19.271.32, p<0.0001) (Figure no.
5). Administration of apidiet+RJ (lot II) led
to the significant statistic decrease of betaglobulin in relation to the control lot
standard food (lot I) (26.470.44 versus
18.320.26, p<0.0001) (Figure no. 5). No
significant statistic differences between the
studied lots for gamma-globulin were
registered (Figure no. 6).
Albumin/Globulins.
Coadministration of acrylamide and apidiet
(lot III) determined the significant statistic
increase of albumin/globulin ratio, in
relation to the control lot with standard
food (lot I) (0.430.01 versus 0.720.04,
p<0.0001) (Figure no. 7). Administration of
apidiet+RJ (lot II) led to the significant
statistic increase of albumin/globulins ratio
in comparison to the control lot receiving
standard food (lot I) (0.430.01 versus
0.680.03, p<0.0001) (Figure no. 7). Little
information regarding the acrylamide
toxico-kinetics in humans is available.
Symptoms described after the intoxication
with highlighted the fact that it is absorbed
in human body by ingestion, cutaneous or
inhalator way (***IPCS.1985, ***
European Risk Assessment).

Results and discussion

We mention that, in the present study,
the control lot that received acrylamide did
not survive without treatment; so that all
the results obtained on the lot treated with
apidiet+RJ (lot III) have a high
significance. A killer rat was observed in
the lot with acrylamide+apidiet, this
behaviour, probably, due to acrylamide
neuro-toxicity, associated with a genetic
factor; it killed and partially devoured 6
cohabitants and it dead after that, by toxic
overdose. Due to this fact, we eliminated
this lot from the study. We compared the
results obtained in acrylamide+apidiet+RJ
lot with results from the two control lots
the control lot with standard food and
apidiet+RJ, respectively.
Co-administration
of
Albumin.
acrylamide and apidiet (lot III) led to a
significant statistic increase of albumin
level, comparatively to the control lot with
standard food (lot I) (30.140.69 versus
41.71.34, p<0.0001) (Figure no. 1).
Administration of apidiet+RJ (lot II) led to
a significant statistic increase of albumin
level, in comparison to the control lot with
standard food (lot I) (30.140.69 versus
40.710.95, p<0.0001) (Figure no. 1).
Globulins. Co-administration of
acrylamide and apidiet (lot III) determined
a significant statistic decrease of globulins
level comparatively to the control lot with
standard food (lot I) (69.850.69 versus
58.411.12, p<0.0001) (Figure no. 2).
Administration of apidiet+RJ (lot II) led to
a significant statistic decrease of globulins
level, in comparison to the control lot with
standard food (lot I) (69.850.69 versus
59.141.06, p<0.0001) (Figure no. 2).
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Figure no. 1 Average values of albumins and

standard deviation (* a p<0.0001 vs. control lot
standard food).

Figure no. 5 Average values of beta-globulin and

standard deviation (* a p<0.0001 vs. control lot
standard food).

Figure no. 2 Average values of globulins and

standard deviation (* a p<0.0001 vs. control lot
standard food).

Figure no. 6 Average values of gamma-globulin

and standard deviation.

Figure no. 7 Average values of albumin/globulin

ratio and standard deviation (* a p<0.0001 vs.
control lot standard food).

Figure no. 3 Average values of alpha-1 globulin

and standard deviation (* a p<0.0001 vs. control
lot standard food).

Information
about
absorption,
distribution, metabolization and excretion of
acrylamide in rats is numerous. Experiments on
rats showed that the absorption of acrylamide can
be orally, or cutaneous or by inhalation (Sumner
S. & colab. 2003). Acrylamide is metabolized in
two ways: by conjugation with glutathione or by
oxidation to glycidamide, a toxic epoxide
(Fennel.T.R.&colab., 2003). Direct conjugation
of acrylamide with glutathione leads to N-acetylS-(3-amino-3-oxopropil) cysteine and S-(3amino-3-oxopropil)-cysteine, which are excreted

Figure no. 4 Average values of alpha-2 globulin

and standard deviation.

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in urine (Sumner S.& colab. 2003, Friedman

M.,2003). Glycidamide is excreted in urine also,
and, after that, gets through the same way of
conjugation with glutathione, leading to N-acetylS-(3-amino-2-hydroxi-3-oxopropyl)cysteine and
N-acetyl-S-(1-carbamoil-2-hydroxi-etil)cysteine,
which are excreted by renal way, also (Fennel
T.R.,.&colab., 2003). The main iso-enzyme
involved in the acrylamide metabolism is
cytochrome P450E1 (Park, J.&colab., 2002); the
other cytochromes do not metabolize acrylamide
in the absence of P450E1 (Sumner S.&colab.,
2003).
In April, 2002, researchers from the
Swedish National Food Administration and
Stockholm University announced the presence of
acrylamide in various foods, alerting the whole
world, because of the well-known toxic profile of
this compound. Many countries were involved in
an international program with the following
objectives: to develop sensitive methods of
quantitative analysis of acrylamide, to determine
the toxic concentration in different foods, to
discover its mechanism, and possibilities of
reducing and preventing the presence of
acrylamide in foods (Lindsay, R., 2002).
The results of this experimental study are
in accordance with those cited in specialty
literature. Rodica Cuciureanu and collab. noted:
beginning with the first day of experiment, the
animals behaviour was completely modified.
Animals in the lot treated with acrylamide
presented specific symptoms of neuro-toxic
effects of it: weight loss, depression, trembling,
weakness in extremities. These phenomena were
absent in lots treated with bee products, and
animal behaviour was almost similar with the
control lot; exception, an apathy state of 30-40
minutes after the toxic administration.

In conclusion, products such as Apiregya,

ApiImunostim, ApiImunomod can be efficient in
liver affection, but recommended only attending
to the results of serum proteins electrophoresis.
Treatment also should be modulated during the
evolution.
.

References
Andritoiu, V. Mierea, In cap VI. Produsele apicole,
Apiterapia n planningul familial, tez de disertaie,
pp. 162-166, Arad, 2006.
Cuciureanu, R., Bulea, D., Cuciureanu, M., Prunel, A.,
Acrilamida din alimente. Compus cu actiune
potential cancerigen. Rev. Med. Chir. Soc. Med.
Nat., 109, 4, suppl. 1, 402-412, 2005.
Cuciureanu R., Andrioiu, C.V., Vrlan, I., Cuciureanu,
M., Murariu, M.: Rolul unor produse apicole n
diminuarea toxicitii acrilamidei, lucrare susinut la
al II-lea Congres de Apiterapie, Iai, 2008.
Eriksson, E., Acrylamide in food products:
Identification,
formation
and
analytical
methodology. Doctoral Thesis, Stockholm
University, 2005.
Fennel, T. R., Snyder, R. W., Krol, W. L., Sumner, S.
C. J., Comparison of the Hemoglobin Adducts
Formed
by
Administration
of
NMethylolacrylamide and Acrylamide to Rats,
Toxicological Sciences, 71, 164-175, 2003.
Iliesiu, N.V.: Apicultura contemporana se orienteaza
spre diversificarea productiei apicole si rentabilizarea
ei; Un potential apicol cu perspective de valorificare:
larvele, in Apilarnil, Editura Apimondia, Bucuresti,
pp.23, 1991.
Lindsay, R., Generalized Potential Origins of
Acrylamide in Foods, Workshop on FRI
Acrylamide Project, Chicago, 2002.
Mrghita L.A.: Albinele si produsele lor. Cap.
Produsele apicole si principalele lor insusiri. Subcap.
Mierea, pp. 280-326, Ed. Ceres, Bucuresti, 2005.
Park, J., Kamendulis, L. M., Friedman, M. A., Klaunig,
J., Acrylamide - induced cellular transformation.
Toxicol. Sci., 65, 177-83, 2002.
Sumner, S., Williams, C. C., Snyder, R. W., Krol, W.
L., Asgharian, B., Fennel, T. R., Acrylamide: A
Comparison of Metabolism and Hemoglobin
Adducts in Rodents following Dermal,
Intraperitoneal, Oral, or Inhalation Exposure.
Toxicol. Sci., 75, 260-270, 2003.
***, CERHR, Monograph on the Potential Human
Reproductive and Developmental Effects of
Acrylamide, 2005.
*** NICNAS, Acrylamide. Priority Existing
Chemical. Assessment Report No. 23, 2002.

Conclusions
Administration
of
standard
food
determined a decrease in albumin level and
increase of globulins values (alpha-1, alpha-2,
beta and gamma), leading to liver affection risk.
Administration of apidiet+RJ in lots with previous
intoxication with acrylamide maintained normal
values of albumin and globulins (alpha-1, alpha-2,
beta, and gamma). Values are comparable with
those in healthy control lots receiving apitherapy.
116