Annals of RSCB Vol. XIV, Issue 1

112

RESEARCH ON API-DIET INFLUENCE OF LIPIDIC PROFILE IN

WISTAR RAT SUFFERING FROM HEPATOPATHY INDUCED BY

ACRYLAMIDE

C.V. Andrioiu1, V. Andrioiu

2, Magdalena Cuciureanu

3,

Rodica Cuciureanu4, Cristina Mihaela Ghiciuc

1, Liliana Taru

1,

Alina Manole5, Ctlina Elena Lupuoru

1

1DISCIPLINE 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

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

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

Annals of RSCB Vol. XIV, Issue 1

113

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, anti-

oxidant, 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, 1991

3).

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).

1The 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.

3Chapter 1: Contemporary apiculture turns to

diversification of bee production and its

capitalization. Subchapter 2: A bee potential with

profit perspectives: larvae, page 23.

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 experimen-

tally 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

4The society office and laboratories are located in

Blaneti Parish, Gorj County, Romania, phone:

0253270221.

Annals of RSCB Vol. XIV, Issue 1

114

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.

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.

Albumin. Co-administration of

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

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115

Figure no. 1 Average values of albumins and

standard deviation (* a p

Annals of RSCB Vol. XIV, Issue 1

116

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-acetyl-

S-(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.

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.

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 N-

Methylolacrylamide 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.