Translation 2579

FISHERIES RESEARCH BOARD OF CANADA
Transl-ation Series No. 2579
Biological evaluatiôn ôf the nutritive value of edible fats. Part II.
Effect of selected fats on changes in the blood serum.lipid fractions
and the levels of fatty acids and of their trans isomers in the.
tissue lipids of some organs
by Swiatoslaw Ziemlanski, Barbara Bulhak-Jachymczyk,
Barbara Kucharczyk, Malgorzata Rusiecka,'Teresa Opuszynska,
Janina Budzynska-Topolowska, Alina Piekarzewska; Augustyn
Jakubowski,-and Piotr Krasnodebski
Original title: Badaniabiologiczne wartosci zywieniowych
• tlùszczow jadalnych. Czesc Ii. Wplyw wybranych tluszczôw na •.
zmiany frakcji liidowych surowicy krwi i zawartosc kwasow
.tluszczowych i ich izomerow trans. w lipidach tkanek
wybranych narzadow
Roczniki Panstwowego Zakladu Higieny (Annals of the State
From:
Institute of Hygiene), 23(5) :.575-591, 1972
Translated by the Translation Bureau(JN0)
Foreign Languages Division
Department of the Secretary of State of Canada
Departrnent of the Environment'
Fisheries Research Board ofCanada
Halifax Laboratory
Halifax, N. S.
:Z"-1 pages typescript
..1.011M•11.010.10•1•111
•
•
•
-I(
1-DEPARTMENT
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*lee
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f-RANSCATE0 FROM - TRADUCTION DE
AUTHOR
English
Polish
- AUTEUR 'Swiatoslaw nàmlalIski,
Barbara Bu/hak-Jachymczyk, Barbara Kucharczyk,
laIgorzata Rusiecka, Teresa Opuszynska, Janina Budzynska-Topolowska,
Alina Piekarzewska, Augustyn Jakubowski and Piotr Krasnodeski.
TITLE IN ENGLISH - TITRE ANGLAIS
Biological evaluation of the nutritive value of edible fats. Part II.
The effect of selected fats on changes in the blood serum lipid fractions
and the levels of fatty acids and of their trans isomers in the tissue
lipids of some or gans.
TITLE IN FOREIGN LANGUAGE (TRANSLITERATE FOREIGN CHARACTERS)
Badania biologiczne wartoè'ci ywieniowych
TIIRE àNzL.Neg T RASecralriAiNyS giR E. EN CÔRnT ktà.5
S
Wp1yw wybranych tluszcz6w na zmiany frakcji
lipidowych surowicy krwi i zawartoU kwas6w tluszczowych i ich izomer6w trans
w lipidach tkanek wybranych narze6w.
REFERENCE IN FOREIGN LANGUAGE (NAME OF BOOK OR PUBLICATION) IN FULL. TRANSLITERATE FOREIGN CHARACTERS.
RÉFÉRENCE EN LANGUE ÉTRANGÉRE (NOM DU LIVRE OU PUBLICATION), AU COMPLET, TRANSCRIRE EN CARACTÈRES ROMAINS.
Roczniki Palistwowego Zak/adu Higieny
REFERENCE IN ENGLISH - RÉFÉRENCE EN ANGLAIS
Annals of the State Institute of Hygiene
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LIEU DÉ PUBLICATION
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1972
FisheriesService
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575 - 591
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J. No .
MAY 2 81973
Roczniki PaAstwowego Zakxadu Higieny (Annals of the State Institute of Hygiene), 1972, Vol. 23, No. 5• pP• 575 - 591.
Biological evaluation of the nutritive value of edible fats.
II.
The effect of selected fats on changes in the blood serum
lipid fractions and the levels of fatty acids and of their
trans-isomers in the tissue lipids of some organs.
By:
Swiatos-law Ziemla6ski, Barbara Bulhak-Jachymczyk, Barbara
Kucharczyk, 101a-lgorzata Rusiecka, Teresa Opuszynska, Janina
BudzyAska-Topolowska, Alina Piekarzewska, Augustyn Jakubowski and Piotr Krasnodgbski
From the De.par.tmént of Nutritional Biochemistry and Physiology of the Institute of Food and Nutrition in Warsaw,
Head of Department : Dr. 9. Ziemlanski D'I.D.,
and from the Institute of the Fat Industry in Warsaw,
Director:
sos-200-10-31
Dr. A. Jakubowski.
UriEDtTtD TRANSLATION
Fer infonna^ion only
Ti2taDi1CTi::N NON REVISEE
Inforimalrion seulement
2
Long-term experiments on Wistar strain rats were
conducted to study the effect of applying in their diets
liquid and hardened rapeseed oil, as well as experimental margarines of various raw-material composition and
of different erucic acid content, on changes in the
lipid fractions of the blood serum, on the composition
of fatty acids and the levels of their trans-isomers
in selected tissues.
In the first part of this study (1) therewere presented
the general and methodological basis for the experiments conducted and also the results of some physiologico-biochemical
and histochemical experiments.
The aim of the present section
is to present further results of tests on the effect of the
application in the diet of experimental margarines and rapeseed
oil on changes in the lipid fractionsof the blood serum and
the content of the lipid fractions and fatty acids in selected
organs and tissues (heart, liver, lungs, adrenals, subcutaneous
and perirenal fatty tissue).
Determinations were also made of
the content of trans-isomers of fatty acids in the lipids of
the lungs, heart, subcutaneous and perirenal fatty tissue, and
also in the phospholipid e triglyceride and cholesterol ester
fraction of the liver.
Four kinds of margarine (MA, MB, MC, MD), of a different
raw-material composition and with differing fatty acid content
were employed in the experiments.
Butter (M) was employed in
the control group. The erucuc acid content in the test margarines ranged from 18% (MA) to 33% (MB).
In parallel to these
studies there was conducted a year-long experiment with liquid
rapeseed oil (OR), containing 47% erucic acid, and a 6 month
experiment with hardened rapeseed oil, the so=cal-lp^1 ;'@ma;,' fat.
The animals received, ad libitum, the experimental die_ts which
5-76
contained 15% by weight of fat, comprising the margarines, the
liquid rapeseed oil, the "0ma" fat or butter as the porntrol fat.^
One group of animals received only a rearing=gr-.anu-lated diet,
from the "Bacutil" firm, containing 7% calories from the fat.
.The composition of the fatty acids in the diets employed is
illustrated in Table I.
In the lipids of the blood serum s.nd of the tissues of
selected organs there was determined the 1eve1 of total lipid
(2), phospholipids (3) and of total cholesterol (11.). In the
tissue of the liver, lungs, heart, perirenal and subçutaneous
fat there was d.etermined the content of total fat (2, 5) and,
except for the perirenal and subcutaneous at tissue, the content of cholesterol ( 4).
The method of Fplch !^t al. ( 6) was
used for the extraction of lipids from the liuer, heart, lung
and adrenal tissue. The blood for testing was.eolleçted in
vivo from the tail after 3, 6 and 9 months duratiôn 9f the experiment, and also after decapitation of the animals after 7
and 12 months.
CHANGES IN THE LIPID FRACTIONS IN THE BLOOD
..^_ SEEIIM
..__._..
No significant differences were found in oomparing the
results of determinations of the total fat and of the phospholipids in the blood serum in groups M&, MB, VC and NID with respect to group P.: in the in vivo tests after 3, 6 and 9 months
of the experiment and also after decapitation of the animals
after 12 months of application of the diet (Figure 1). After
12 months there was found an increase in the level of total at
577
I
I
Table I
The fatty acid composition of the tested-fat s
Tabela 1
SkIad 1,,tvasôw tliiszevo-,~-3-eh badanveh fluszcz6w
Ir
__J . 1. ,
C16 :0
1 C6 : 0 C'- :O C
! pal .
.1
rni- .i
Icap . kap- kap- îi lau- ~I
rysrone- i rylo- ryno . ryno - ;
no- ;
W3~ ~ , wy M'y I wy !
Margaryna A MA
Margaryna B MB
Margaryna C Y, C
Margaryna .D
IYM
Maslo
M
Olej rzepalzoivy OR
,,Oma "
- I -
I'l
0,05
0,05
01 01
0 :2 0:2
0,8 1,9
- 70,2 0,2
0
C18 :t
0,2
0,4
0,9
1,4
2,7
O'l
2,0
1 C18 :2
C14 .1
pal
., 1 Ini .
C15 :0
"- i to-o-
ty no .
i
%vy
0,4
0,5
1,8
1, 4
9 lejo .
W.%
.
5,5 1 0A
55 ( 0,5)
10:0 1
12,2
O'l
1,3
10,8
38,0
3:4
62
1,8 1 1, 9
.2
0,6
2, :3
0,2
0,4
C32- 0
C"* : lino . ~ ejkolino- ~ 1 belie- 1 cru .
ole- 1 iizo~ Tyno-!
leno- 'zenostea-
C17 :0
1
linowy 1 mer I loivy 1 .,%Vy 1
i
1
i
Margaryna A
hIargar~ na B
Margaryna C
Margaryna D
Mash?
Olej rZepakowy
1,0
5,0
4,2
4,6
4,3
9,2
1,2
3, 5
.i
i
2,1 i 22,6
1,4
32,3
23,4 2,1 1 19,1 1,5
1,3
22,7 . 1,7
25,4
23,8 1,9 23,1 1,1
22,7 1 , 8 1,5
14,9
8,1
13,2 1,8 15,9 2, 9
20,1
nowy i 1,
Nvy
i
I
8,9 2,5 , 18,4
6,3 3,0 1,,33 .1)
5,5
2,1 1 20 . 3
2,2 1' 22 .9
5, 0
7,8 0,8 47 .6
5,9 2,5 37 .1
I
c6to
c8 : 0
C1010
C1210
C14. : 0
C14- :1
C15 : 0
n6to
016tl
C17 0
C1 8 0
C1 8 1
C18 :2
C18 .-2
C1 8 st 3
C20 :1
C22 : 0
C22 i I
caproic
caprylic
capric
lauric
myristic
palmitic
palmitolei c
stearic
oleic
isomer
linoleic
linolenic
elcoseenic (gadoleic ?)
beheni c
erucic
• à
5
Total fat
•0 g 500-
1".
T1zi.‘7r7 calkomty
■ Jk.•
• ,4opi
-e)
•-P
31,01,-
E " ` 200 1-
•JD, celocLL
_L
3
L-40
E mg%
à
12
Nargaryna A
11
8
Phospholipids
300L
Fosfolipidy
------ .•
Bdter
■
200r-
,
100
à
Cholesterol caiRowity
r
•
-----_ _ • •
.00
•
•ea
•
•
Total cholesterol
mØg
100
12
'
.
3
à
,
• -
-------9—
-
,
n
•
-Pliesifice trecama daskraciczenia
Mont
s oi auraIlon of experiment
'Rye. 1. Poziom frakcji lipidowych w surowicy lorwl.
Figure 1.
The level of.lipid fractions in the blood serum.
in all of the groups of rats which received the margarines, as
the control group which received butter in its diet.
comparedt
Statistically significant differences were evident between
group M and groups MB, MC and MD.
The level of phospholipids in the serum of the rats
(Figure 1) after 12 months was higher in groups MB, MC and MD,
as compared with group M, and in this case a significant difference was found between group M and groups MC and MD.
6
Deserving particular attention are the very considerable
seasonal differences Which we found in the level of the tested
lipid fractions in the blood serum.
As is evident from our
tests the level of the total fat and of the phospholipids in
the blood serum is higher during the period from April to September, and moreover the highest values were obtained in June.
This finding possesses not only a real theoretical significance
but also a practical one, since it points to the necessity of
being careful in interpreting and comparing results obtained at
•
different times of the year.
A statistical analysis of the total cholesterol level
in the blood serum after 12 months of application of the diet
indicated that the cholesterol content was lower in group MA as
compared with the group of animals that received butter in their
diet, whereas in the remaining groups of animals which received
margarine the cholesterol level in the blood serum was also
somewhat lower as compared with the group that had received
butter, though these differences were not statistically significant. After 9 months of the experiment, however, the choleste-
rol values in group MA were higher as compared with the other
g roups .
CHANGES IN THE LIPID FRACTIONS IN SELECTED ORGANS
In the liver tissue after 7 months of the experiment the
level of total fat in the groups that received margarine in their
diet was lower or equal to that of the group receiving butter,
and in this case statistically significant differences were demonstrated between group M and groups MA and MB.
Similar
7
changes in the total fat content were also noted in the lung
tissue.
However, statistically significant differences were
found only in groups Prr!A and MC.
After 12 months of the experiment the total fat content
in the liver anf lung tissue was higher in the animals that
received margarine, with the exception of group MB, as compared
with the animals which were given butter in their diet (Figure 2).
In the heart muscle, on the other hand, in the groups
of animals that received margarine in their diet over a period
of 12 months the total fat level was almost the same (groùp T4îC)
or higher than in the group of animals that received butter,
and in this case the difference between group BOD and group M
In the group which received
was statistically significant.
butter the total fat content averaged 13.7 ±0.4 mg / 1 g tissue,
while in group ND this was 26.4. ±1.4. mg/ 1 g tissue. The
data obtained are illustrated in-Figure 2.
wqtroba
Qtuca
Liver
i,ungs
Serce
Heart
fi
..s0
I
4
30
k. :
20
M.
10
,..
I
0 -,.j
N
M-A - margaryna A
M-8 - - „ - B
-C - - u -- C
M-°nasto Butter
p 7 miesiqcu s►osowania diety
12 -n^ - n-. -u-
mQnths of application of
diet
Ryc. 2. Poziom t1uszczu calkowitego w przeliczeniu na kwas palmitynowy w tkan-
+kach.
Figure 2. The level of total fat, calculated as palmitic acid,
in tissues.
-8
11
6
I
Q2'
211
_R
0
ÇI)
ti
Ryc . 3. Poziom- cholesterolu calk-owitego w nadnerczach po 12 miesiqcacb dogwiad-,
czenia .
Figure 3 .
The level of total cholesterol -in the adrenals after
12 months of the,experiment .
Of particular interest is the cholesterol level in the
adrenals . In the adrenal tissue after 12 months of the experiment the cholesterol level in the group of animals which received
margarine B in their diet comprised 11.8-5 ± 4 .9 mg per 1 g tissue
and was considerably higher than that of the group which receive d
butter in its diet or even those which received the other margarines . We found particularly large amounts of cholestero l
in the adrenals after giving the animals in their diet the liquid
rapeseed oil, containing about 11 .7% erucic acid . The cholesterol
content in the adrenals of this group averaged 80 .8 mg/ 1 g tissue
(Figure 3)-
In studying the effect of the experimental margarines
on the total fat and cholesterol level in the selected tissues
it was found that these margarines did not have any substantial,
constant effect on changes in the above mentioned parameters,
with the exception of margarine B which had the highest content
of high erucic acid rapeseed oil.
This effect is most clearly
apparent in the adrenals, which may be accepted as a very characteristic disturbance connected with the erucic acid content
Ln the diet (7, 8, 9, 10, 11).
THE FATTY ACID COMPOSITION OF THE TISSUE LIPIDS *
The effect of fats in the diet, with different content
of liquid and hardened rapeseed oil, on the fatty acid composition of the tissue lipids has for several years been a subject
of study by many authors (12, 13, 14, 15, 16, 17, 18, 19, 20,
21, 22, 23, 24).
The fatty acid composition was determined by the method
of gas-liquid chromatography in the lipids of the following
tissues:
heart, liver, liver mitochondria, lungs, subcutaneous
fatty tissue and perirenal fatty tissue. For the determinations
we employed a Pye gas chromatograuh with a flame ionization detector.
From the data obtained it is apparent that the greatest
group differences in the fatty acid composition were found in
the perirenal and subcutaneous fat tissue.
These differences
* In the present paper only the general changes in the fatty
acid composition of the selected organs are discussed. The
detailed analysis of the fatty acid composition of the lipids
of the various tissues will be the subject of a separate publication.
10
a m Kwasy nasycone
b f-7-1 Kwasy jednonrenasycone
C EM Kwasy ivielonlenasycone
After 7 months
Pa 7 mie: rqcach
°l 60 60..4O 30 20 10
Sümy kwâsôw
After 12 months
!
I
Pd 12 miesiqcach
10 20 30 40 50 60
^- Sumy kwasôwAC].d
frodowlE^
_^
I
totalB
Stoçk•
(rearing diet )
Rye. 4., Procentma zavtTartosc kwasdw tluszczowych nasyconych, jednonienasyconych i wielonienasyconych w lipidach tkanki tluszczowe5 r-odskbrnei.
Figure 4. The percentage composition of saturated, mono-unsaturated and poly-unsturated fatty acids in the lipids of the subA
cutaneous fat tissue.
a - saturated acids
b - mono-unsaturated acids
c -- poly-unsaturated acids
become greater as time passes. In the more metabolically active
tissues, as for example in the tissue of the heart, liver, liver
mitochondria and lungs, the differences are smaller and become
even more obliterated as the experiment proceeds (Figures 4, 5
and 6).
The small content of saturated fatty acids in the tested
margarines clearly changes the value of the ratio of saturated
to unsaturated fatty acids in all of the examined tissues, and
11
.
_
____ ... _ . ..-_.______
7' 6 5 4 3 2 1
-
.
7
'1 ,.
! ?
•
Tk ttuszcz poaskarnaSu
. b cutaneous
Perirenal.6.tius.tÈ7.okok,, ner.kowa
fat
tissue
.
1.=.7_,
7.,110dom22
.
0
fat tissue
tias;iO3
.
, n,
.
• .:•=4 IN-A
i
irf".3
1
•
.=
. ■
. .
. ... ..
Watroba
..
Liver
e4-8 Igns.
1"1 - c 'r- 7
•
• .
t
.
1
i,
.
ti-D 'Mr•gn
r•='! OMA . rt
. , OR
. ..
e"
.
,
—
'rel--
' :,
.•
. .. .
•
.
4
-
1
Mitochondria wcitrobç
IIver mitochéndria
.
mashr--7.1----butter
•
ilocirmq
M-C
. . .477'71 M-D .°737-1
OMA
OR
.M.]
Serce
He art
•
Nodow
tiasto
M-A
•
8
•
Stock (rearing diet)
Kwasy nasycone • a. .
b
Kwasy nienasycone
po 7 mostacach
Kwasy nt:enasycone c
po 1Z miestarach
-f:
;
-
eunlOCIA
.Rye. 5. 15roporeja kwas6w tluszczowych nasyconych i nienasyconych go 7 i 12
raiesiqeach poda•wania ,diety (sum:: kwas6w nasyecnych przyjgto za jednostkg). .
Figure 5. The proportion of saturated and unsaturated fatty
acids after 7 and 12 months of application of the diet (the
sum of the saturated acids was taken as unity)..
b c a
Saturated acids
Unsaturated acids after 7 months
Unsaturated acids after 12 months
especially.in the subcutaneous and perirenal fat tissue, i.e.
in the so-called reserve fat (Figures 4 and 5).
After 12 months
of the experiment this ratio in the MA and MB groups of animals,
for example, averaged
1 : 6.3
(Figure 5).
12
From the studies of many authors it is known that the
most favourable ratio should be
1 : 2.3 (= 0.43). Thomasson
(25) correctly emphasizes that many natural oils and fats do
not provide the most favourable composition of fatty acids for
the organism.
On making an overall evaluation of the results obtained
it may be stated that the greatest differences in the fatty acid
composition of the lipids of the tested tissues, inthe group of
animals receiving margarine as compared with the group receiving
butter, relate to the accumulated amounts of saturated fatty
acids, especially palmitic acid, and also to the poly-unsaturated
acids.
The . tested margarines provide too small an amount of •
saturated acids, they are however a good source of the polyunsaturated acids, because of the fact that they contain 30%
unhardened sunflower oil.
In all of the tissues examined there was found a lower
content of palmitic acid in the animals which received margarines
in their diet, as compared with the group of animals which received butter in the diet (Figure 7).
In the fat of the diet
which contained butter there was 32.7% palmitic acid, while in
the fat of the diets containing the margarines this ranged from
6.6
to 12.4%.
The accumulation of linoleic acid in the tissues
of the rats shows a dependence, similar to that of palmitic
acid, on the level of this acid in the dietary fat (diet MI 5.1%,
diets MA, MB, MD: 22.3% and diet MC: 26.1%).
Among the tested
organs the highest percentage contribution of linoleic acid was
found in the lipids of the heart and of the liver (23.2 - 27.7%)
(Figure 8).
13
.
.
1
..
.
..
.
a um *miry nmycone
b. EM Kwasy jednonienasycone
o I= Kwasy wielonienasycone
-
.
After 7 months . After. 12 months
.
•
•
.
'
Po 12 rmesiacach
W 20 ;30 •<.p., ,5p 6•0%
..4!mY Kwaidw Ac
'
Po 7 miesieach
0460K5040 3p 20 W
'
Surrig' 'kwa'selw
Hnclowl
MELIZSMTEINSIffil
MUM
CC=913111311
Mask)
M-A
73e125117J- 517
id tota l
Sto«
(rearing diet)
Butter
r3ZISINTM
M- 13
M-C
M-D
OMA
.
=•,
_ • .. .
'
Bffegemian
OR
.
I,..
Ryc. 6. 15rocentol.va zawartogé kwaselw tluszczowych nasyconych, jednonienasyconych 1 wielonienasyconych w lipidach serca.
Figure 6. The percentage content of saturated, mono-unsaturated
and poly-unsaturated fatty acids. in the lipids of the heart.
• a - Saturated acids
b - Mono-unsaturated acids
- Poiy-unsaturated acids
Comparing the composition of the fatty acids of the
liver mitochondria after 7 and 12 months of presentation of the
diet it may be stated that with the passage of time there occurs
a change in the proportion of saturated to unsaturated fatty
acids, in the direction of lowering the amount of the latter
(Figure 9). This change is brought about by both a decrease
in the content of the poly-unsaturated acids as well as by a
certain increase in the content of the saturated fatty acids,
which is particularly marked in the group of animals receiving
butter.
L.
'
Afot^2rm,^2^^m^onths
After 7 mànt^is
t '% 7 m,est4car
5 10 15 20 25 30
30 '25 20 45 10 5
^
k wasow
tka tu^cznwa okofnnerkowa kfaC1Ug
a,.,Tkan
t>^RLTMR . -=c.ts;
•
f_.
^Hadowl
:^s•,z;^ Masto
M-A
CLS:•
3>^ 7M-C
M-D
L?:7^:F7kG
CMrn.CXR!C( OKA
OR
0f
1. . _.. ,^.,•.^yara^
`
' A
•D
L'^s7^
^
b. Tkanka tfius<czowa podskflrr,a
fdi>tïa^^J&E^
^^y
%^ How7:^I
rva^ÿ ^??sn1 Masto
M-A
.t37
t;^ w^•^ M-C
.^taTl
M-D
OMA
MrUMQ.4
OR
C, Serce
Hadowl. Imr-,V3rB
Masfo
-'U
°y'it
M-A
M-B
^.. M-C tF^m3.:rm9
Ce>:
tr^:^ =
=
M-D
ÔRA
fw '^
émoa
d. Wntroba
Hodowl.
Ftasio.
14 -A
Irw^e•a.^s7 M-8
.Wr^1 ^. LMFM,
Cr..
V:I.:;
^5^;`..,'.ycY7^
,.t
M-C
QNA
OR"
n
e . Mitochondria watmby
Hoiowt
Mas?o tk°' ' C
M-A
PI-B Gi-n-=s'i,<.aïJ
YT•lc3•J537
.
M-C •R^.:.`
.-=:SiV•..'E4f
ly-D
^ OORA
.
*f^Po12 m,esiacach nie oznaczano
Rye. 7. Zawarto§é kwasu polirnitynowego w lipidach tkankowych.
Figure 7. Content of palmitic acid in the tissue lipids.
a.
b.
c.
d.
e.
=
-
Perirenal fat tissue
Subcutaneous fat tissue
Heart
Liver
Liver mitochondria
# Not determined after 12 months
In the lipids of the liver after 12 months of application of the diet, in comparison with the data obtained after 7
months, there occurs a change in the proportion of saturated to
unsaturated acids, in the direction of increasing the amount of
unsaturated acids (Figure 10). The character of this change is
the reverse of that in the case of the lipids of the liver
15
After 7j.qpnths AfP0t-eFm12 . mcopths
Po 7 miesiecy lesloca
c
4L- 42 25 20 15 10 5 5 10 15, 20 25 30 36
a .Tkanka ttuszczowa okoton'erkow~
K~vasow
Podawl
ni Mwto
H-A
M- C
M-D
OMA
OR
0MV-VMMMwMC5~=
b .rkanka ttuszczowa podskdrna
Hodowl ks
Masto
M A
WK,
OM A
OR
c
Pt
SSLOrCL,
Modowl
M-A
M-B
M
OIYA
OR
d . W4froba
- - x
F1
.
.
Hodowl.
Mask
M-A M=,
"z-_-An
OR
esMitochond6o wqtroby
Hoaowl.
M-A
OMA
OR
k=. :~ E,
XFA M
4 P0 12 miasic;cach nie oznaczano
.8 . Zawartogd kwasu lirlolmvego w lipidach tka-nkowych .
Figure'8- Content of linoleic acid in the tissue lipids .
a.
b.
c.
d.
e.
-.
Perirenal fat tissue
Subcutaneous fat tissue
Heart
Live r
Liver mitochondri a
Not determined after 12 month s
mitochondria (Figure 9) .
In the groups of animals that received
margarines 'In their diets the change in this proportion results
primarily from the decrease in the amount of saturated acids .
In the animals that received butter in their diet there
occurred a decrease in the content of saturated acids and an
.16
increase in the content of poly-unsaturated acids, probably as
a result of the decrease in the total fatty degeneration of the
liver tissue after 12 months. It should be noted, however, that
in this group of animals the total content of saturated acids
in the lipids of the liver, in comparison with the margarine
groups, is higher after 7 as well as after 12 months.
On the
other hand the content of poly-unsaturated acids is smaller. •
Comparing the content of saturated and poly-unsaturated
fatty acids in the tissue lipids of the heart muscle with the
content Ln . the lipids of other organs, such as, for example,
the liver and lungs, it should be assumed that the composition
of the fatty acids of the heart lipids is to a certain extent
conditioned by the content of the fatty acids in the diet
(Figures 5, 6, 7, 8).
This is connected with the fact that the heart muscle
to a considerable degree exploits the fatty acids of the diet
as a source of energy. Erucic acid displays
a particularly
great dependence between the content in the diet and the content in the tissues.
Therefore particular attention was paid to the content
of this long-chain fatty acid in the analysis of the determination of the fatty acid composition of the tissues (Figure 11).
It was found that erucic acid is accumulated mainly in
the subcutaneous and perirenal fat. The highest content of
erucic acid in the tissues was obtained in the animals winch
received margarine B in their diet, i.e. the margarine containing
the highest amount of this acid (33%); eg.
in the subcutaneous
fat there was found after 7 months 4.4% and after 12 months 3.7%,
17
.
a. = Kwasy nasycnne
b. C:I .Kwasy /édnonienasymne
C. n .Kwasy wielonienasycone
•
After.7 months
• PO 7 miesicqcach
60 50 40 30 20 10
,
7t
p
_TT-T-^T7'
After 12 months
Po 12 imesiqcach
40 5() 60'0
10 20 3 0TT-r-
j
Semy kwasow ^ sc^my kwasow -Sum of
acids
^-`^'
xoc^wr
Nasfo
..^+^.;t,^•^._
:..:.:.....:^{ OR
•
:.
Ryc. 9. Procentowa zawartosé kwasôw t3uszczowych nasyconych, j ednonienasyco,nych i wielonienasyconych w lipisiash rnitochondri6w wqtroby.
Figure 9.
Percentage content of saturated, mono-unsaturated
and poly-unsaturated fatty acids in the lipids of the liver
mitochondria.
a. - Saturated acids
b. -• Nono-unsaturated acids
c. - Poly-unsâturated acids
while correspondingly in the perirenal fat these values were
6.1f after 7 months and 4.3% after 12 months. A still greater
dependence was found in the rats that received the high erucic
acid liquid rapeseed oils eg. in the subcutaneous tissue the
erucic acid content was 9.1% after 7 months and 6.4% after 12
months.
For the perirenal tissue the corresponding values were
9.8% and 8.5% of erucic acid.
;
•
1.8
• a-. ell
Kwasy nasycone
b. C3 Kiyasy jednonienasycone
Kwasy wielonienasycone
• e.
After 12 monthà
„After'? months
••
Po 7 miesiacach
.°À;60 e) 40 30 20 10
Surny
Po '12 mieslacach
10
30 40
I
1111=112SZIME
liadowl
SO 60%
kwasnw SUM
ffiZZ:=EEME
of
acids
Mast° ma.
dIFOURIIIIIIIM:1511MRMUMUIM
M-B
"
M-C
IMMEBRUMM
OMA
OR
Rye. 10. Procentowa zawartoàé kwas6w tluszczawych nasyconych, jednonienasycolipidach wqtroby.
nych i wielonienasyconych
Figure 10.
Percentage content of saturated, mono-unsaturated
and poly-unsaturated fatty acids in the lipids of the liver.
a. - Saturated acids
b. - Mono-unsaturated acids
C. - Poly-unsaturated acids
Among the tested organs, the highest accumulation of
erucic acid was found in the lipids of the heart muscle;
eg.
in the MB group of animals - 2.5% after 7 months and 2.4% after
12 months:
In the group of animals receiving liquid rapeseed
oil these values were, correspondingly, 4.8% and 4.2%.
From
these data it is evident that the content of erucic acid in the
tissue of the heart muscle, in contrast to the subcutaneous and
perirenal fat tissue, is not subject to any great variation
19
with the passage of time, i.e. within the limits of the 7 and
12 month period. As is evident from studies recently conducted
by Houtsmuller (19) and also by us (unpublished data), in young
animals the content of erucic acid in the heart muscle increases
in a parallel manner to the degree of fatty degeneration of the
heart and attains its maximal values as early as 3 - 6 days
after the application of the high-fat diet. For example, the
administering in the diet of 50% of the calories from rapeseed
oil leads to a considerable accumulation of erucic acid in the
heart muscle as early as 3 days later; this may amount to27%
to 4.0% of all of the fatty acids. After 4• - 6 weeks of aministering the same quantity of rapeseed oil to the animals, the
content of erucic acid in the heart muscle decreases and attains
a level of about .15 - 20%. At the same time the .st.e.atos.is of
the heart muscle disappears. It may be assumed that this is
connected with the metabolic adaptation of this organ. If however we administer the rapeseed oil in the diet over several
successive weeks, there appear in the heart muscle focal histiocytic infiltrations that become transformed into a fibrosis,
which will be discussed in detail in Part III of our study.
THE CONTENT OF UNSATURATED TRANS ACIDS IN THE TISSUE LIPIDS
The quantitative determination of the trans isomers in
the lipids of the heart, lungs, subcutaneous and perirenal fat
tissue and also in the individual lipid fractions (phospholipids,
triglycerides and cholesterol esters) of the liver tissue was
carried out by measuring the absorption of infra-red radiation
(26, 27).
586
20
As is evident from the data obtained (Figure 12) the
trans isomers of the unsaturated fatty acids had been incorpo7
rated into the lipids of all of the investigated tissues of the
rats which were given margarines in their diet. They were not
found to be present Ln the tissue lipids of the rats which received butter in their diet. Among the tested tissues the largest quantities of the trans isomers were found'in the lipids
of the subcutaneous and perirenal fat tissue (averaging 18.4%
Ln groups MA and MB and about 13% in groups MC and MD). There
was considerably less in the lipids of the lungs (about 11.8%
in groups MA and MB and about 9.0% in groups MC and MD).
.
The
smallest quantities of the trans isomers were found in the
lipids of the heart (averaging about
5.7% in groups MA and MB
and about 4.0% in groups MC and MD).
The individual lipid fractions from the liver contain
differing amounts of the trans isomers.
The largest quantities
of these"were found in the cholesterol esters (averaging 18%
in groups MA and MB and about 16% in groups MC and MD), somewhat smaller values were obtained in the triglycerides (averaging
17% in groups MA and MB and 11.5% in groups MC and MD) and the
least in the phospholipid fractions (averaging 9.6% in all of
the groups).
On comparing the results of the conducted tests it can
be stated that a dependence exists between the quantity of the
trans isomers in the tissue lipids and their content in the
diet.
The quantities of the trans isomers in the lipids of the
tested organs after 7 and 12 months of application of the diet
do not display statistically significant differences in a
21
After 12 months
After 7 7nuesiqcach
months
Po 12 tniesmcach
Po
2 4 6 8 10
. 10 8 6 4 z
%St"Pg a.Tkanka ttuszczowa okoionerkowa al
SUM
k°,asj
u e % acias,
kwasore
lllll
1111
...1111
.
of
. e Hodow ez
""'="
• • »e=f;.:=e ei=e e7‘7.7.7-;re
Ir===i1 11-D esr-unmo
mA
OR
Irtmizuzs•mmus
b.
•
mes=rammommommum
Ticanka thiszczowa podskarna
Nodowl um
Nash, •
`1- A
,rmvuois
:1- A
M - C =uarszw
N-D 'mama
oPtA wzimminiemmi
-i
mitU
ommemassi
' •
weeznacral
•
ir=a
celearermi
trrommeineczre.ffla
•.
DR
- C Sere?
•
Rai Hodowl it
• •
.e
•
•
affli Nast° 1i
eta N- A •
sotoza Pi- B muse
a= N -C km
/I- D
PIme= N-17
erstzd
Mismi
inœr:dtezeig Ooly
d. Wqtrtiba
.•
Hodowl.
11:2s10
M -A
IT?1-C
mu 11-D
um=
NA
wOR
mom
msa
c. Mitochondria
1.
wqtroby
•
Hodowl. 1
Plasio
•
11-C
14-D
OHA
OR
•
• Po 12 iniesiqcach nie oznaczano
Rye. :11. Èmmrto§é kwr—su eru:kowego w lipidaeh tkankowych.
The content of erucic acid in the tissue lipids.
Figure 11.
a.
-
b. c.
-
d. e.
-
Perirenal fat tissue
Subcutaneous fat tissue
Heart
Liver
Liver mitochondria
* Not determined after 12 months
substantial majority of the tested animals; nevertheless, after
as corn12 months the values are, in general, somewhat higher
,
pared to the data obtained after 7 months of administration of .
diet.
the
22
Heart
Serce
Jrkanka thiszczowa podskdma rkanka Ousmowc DkOfO-
0"'j ;~'UUUUUUIIUVUy
J.
zur
Lclu
An r i
' W~V
f at - t LSSU6
Watroba
Liver
Phospholipids . Triglycerides Cholesterol esters
%
Fosfdlipa~ M-5ijIlcerydy rstry' cholesterofa
20~
II
M-A - margaryna
M-B t4-C- .
M-DRyc . 12. Zawartok izomer6w trans wyraiona w
stosowania diety
a r-i 7 mjesi~cy
bM3 12 -- - It- . -$I-
A
8
C
D
Figure 12 . The content of trans isomers expressed as
percentages .
a - 7 months of application of the diet
. of 01 o f
11 of it
b - 12
As an example we present the data from the subcutaneous
fat,tissue in groups MA and MB : after 7 months duration of the
experiment 1?-3% of the trans isomers were found, and this value
was 19 .8% after 12 months . -The quantity of the trans isomers
-in the triglycerides of the liver in groups MA and MB comprised
23
r
16.3% after 7 months, and 18.4% after 12 months.
In the cho-
lesterol esters the differences were smaller and comprised,
correspondingly s 17.4% after 7 months and 18.5% after 12 months.
The phospholipid fractions do not display significant group
differences in the content of the trans isomers.
CONCLUSIONS
The following conclusions may be extracted from the
studies that were carried out:
1. A considerable dependence was found between the content of erucic acid in the diet and th.e total cholesterol content in the adrenals. Especially high quantities of cholesterol
in the adrenals were demonstrated after the administration of
liquid rapeseed oil to the animals in their diet.
2. The fatty acid composition of the diet, administered
over a period of 6, 7 or 12 months, exerts a certain influence
on the fatty acid composition of the lipids of various tissues.
The reserve fat tissue (subcutaneous and perirenal) displays
the greatest dependence in this aspect. In the metabolically
more active tissues such as, for example, the heart, liver and
lungs, the differences are smaller and become even more obliterated as the experiment proceeds.
3. The rapeseed oil and the experimental margarines
possess an'unfavourable ratio of the saturated to unsaturated
fatty acids, which was clearly displayed in the fatty acid composition of the examined tissues. The greatest changes in the
ratio of the saturated to unsaturated fatty acids were found in
the reserve fat tissue, i.e. in the perirenal and subcutaneous
fat.
589
24
I()
VIO
(
4. A marked dependence was found between the content
of erucic acid in the diet and its content in the fat tissue,
i.e. in the perirenal and subcutaneous fat.
Among the tested
organs it was the lipids of the heart muscle which showed the
greatest dependence in this direction.
'
However, the content
of erucic acid in the heart muscle in the long-term experiments
was considerably smaller than that of the reserve fat tissue.
The accumulation of erucic acid in the heart muscle may be the
immediate cause of the occurrence of histopathological changes
in this.
5. There exists a dependence between the content of
the trans isomers of the fatty acids in the diet and their content in the lipids of the tested tissues. The greatest concentration of the trans isomers was found in the reserve fat tissue
(the perirenal and subcutaneous fat).
25
590
r ^t9LS r S ^'AL ^'.AIrTTATTbN TNTSi^ ^ ^7^^ ^ ^^T.F .FATS
ÎI. An influence of selert.ed fats :u,gon the rhang£s in Ufé 1^evè?s •1if fatty
-ag^&5 and of thèir -trans :isome.rs in the tissu lipiF3s aif same,:or.gans
Suimmca-r:y
An influence of iiietary harslened and liquid rape-serd oïl -.^:irl 'of txperimental margarines (MA, MB, Mç, MD) differing in =zm znatex-ial composition and
eontaini3ig various amounts of er=ic acid .upon the. changes in -serum lipids
fractions and on the levels of latty acids in variouu organs (heart, liver, mitochflndria of the :tiver, subcn:taneous _and retro-rean1 fai-ty 'tissues) has been inveatigated during the long-tercn experiments, 6, 7, and -13 momt.fls in duration, using
rats of Wistar strain. The levels of ;erucic acid in the fats under study range d
from 18 to 47 percent, and the contents of at in zli.et covered So percent of caloTic requirements.
Among the most rexnarkable findings was an excessive r3ePnsitïon of cholesterol in adrenals of animals receiving greater amoun;.s of hardened or ïluid rape-seed fli7.. The contents of fatty acids in subcutaàneons and retrL-renal fatty tissue
was 'largely dependent on the composition of dietary 'fai, liver and liver mitochondria being less influenced, -v1th the Jarngs and heart being even less affected by
this factor.
Following a long period of time .('I and 12 months) there was a change in lipid composition in J:ivrr, in tivhich' the proportion of saiurated :Lnsaturated fatty
acids altered on the bènxlf •of the latter group). In mitDchnniria of the liver the
opposite was true i.e. a decrez,;e cf unsaturated and an increase of =aturated fatty
acids. This shift was independent of the fatty acids composition of diet (butter,
margarin or rape-seed oil). The rleposits of ernzi.c acid -were maiidy found in subcutaneous and retro-renal tissues.
In the long-term experiments a great deposition of eruci. ac'd was found in
lipids of the heart. The largest amounts of trans isomers tive^c encontered in fatty
lissue (up to 18 percent). In the heart tissue trans isomers were present in amo- •
unts of about 5 percent. The maximal level of trans isomcrs in the margarines
under study was 42 percent.
591
26
it
PISMIENNICTW O
1 . ~. Ziemlafiski, T . 6puszy?iska, B. Bulhak-jachymczyk, G . Okolska, H
. Bartlet,
. Z. 01szewska, J . Badzyfiska-Topolowska : Badania biologimne -wart09ci 2ywicniowei t1uszcz6w jacialnych . Czqgd I. Roczniki PZH IM9 , 1 23, 387 . - 2. Illinow P. P .,
Vatresni Bolestj (Bulg,) 1967, 6, 367 . - 3. Fiske C. H., Subbarow J. wg J :
Homolki : Diagnostyka biochemiczna wyd . III. PZWL . Warszawa
1961 . - 4 . Bloor
Journ . Biol . Chem . 1947, 170, 671 . - 5 . Bragdon J. H. : Journ . Biol
. Chem, 1951,
i9O, 513 . - 6 . Folch J ., Lees M ., Sloane Stanley G . : Journ. Bloch
. Chem . 1937, 226,
497 . - 7 . Carroll K . K. : Endocrinology 1951, 48, 101 . ~ 8 . Carroll K
. K. : Can . Journ .
Blechem . and Physiol. 1962, 40, 1115 . - 9 . Ro-4ynkowa D ., W. Koziol6wna,
A . Horst:
Acta Physiol, Pol . 1962, 13, 591 . - 10 . Nikonorow M., HoMTOWski S .
: Roczniki
Woj~k . Inst. Hig . EpId . 1968, 3-4, 19 .
,11. Rakowski W . : Poziom cholesterolu w nadnerczach szczur6w iywionych die'A.1 zawierajcqcq olej rzepakowy . Dokumeritacia w Bibliotece Inst . Przern
. Tluszcz,
,w Warszawle 1962. - 12. Hopkins C, J., Murray T. K, Campbell J.: Can
. Journ .
Blochem . Physiol. 1955, 33, 1047 . - 13 . Craig B. M., Youngs C. G., Beare J .
L.,
Cambell- j . - A .: Can. Journ . Biochem . Physiol . 1963 , 41, 43 . - 14. Craig B
. AT.,
Youngs C. G ., Beare J . L ., Campbell J, A .: Can
. Jou m_ Biochem . Physlol . 1963, 41,
51 . - 15. Holman R . T. : Federation Proceedings . 1964, 23, 1062. - 16
. Davis J . T.,
'Bridges R. B ., Coniglio J. G. : Biochem. Journ . 1966, 98, 342
. - 17. Bach'A ., Metals
P., Raulin R ., Jacoquot R. : Bull . Soc. Chim . biol.
1969, 51, 167 . ---7- 18 . Rocquelin G .,
Leclerc J. : Ann . Biol . anim. Bloch. Biophys . 1969, 2, 41-3.
19 . HoutsmuZler U. AT. T . ,
.- Struijk C. B., Van Der Beek A . : Biochim
. Blophys . Acta, 1970, 218, ~ 564 . - 20 .
Beare~ROgeTS J . L.: 1~roceedings of the International Conference on the Science, Technology and Marketing of Rapeseed and Rapeseed Products Sainte-Adele,
Canada (20-23 september 1970) Wyd . w Ottawle 1.970, str . 450.
21 . Beare-Rogers J . L ., Nera F . A ., Heggtveit H. A. : Proceedings p. poz
. (20) str.
450. - 22 . Kodykow B ., Juzewa L., Filipowa G., Kocman Z
., Jakubowski A., Kaczanc;wski Cz.: Prace Inst . Lab . Bad . Przem . Spo2. 1969, 12, 587 . - 23. Rocquelln
G .,
Cluzan R . : Cah . Nut . Diet. (Med . et diet .) . 1971 ., 6, 85. - 24 . Rocquelin G
., Sergiel
J. P ., Martin B., Leclerc J ., Cluzan R. : J6urn . Ain. Oil .Chem
. Soc. 1971, 48, 729 . 25. Thomasson H. J . : International Symposium for the chemistry and Technology
of Rapeseed Old and other Cruciferae . Gdafisk 1967 . ?:eszyty Probl. Podst. N~iuk Rol .
1070, 7- 91, str . 331 . - 26 . Ucciani E. i wsp. : Bull. Soc . Chim . France. 1960, 1511
. 27 . Rapport of the spectroscopy CGmmitte : 1959. Jbqrn Am . Oil Chem . Soc., 627.
Dri . 4-V-1972 r .
Warszawa, ul. Powslfiska 63.
BIBLIOGRAPH Y
1 . S . Ziemlan'ski et al .i Biological studies of the nutritive
value of edible fats . Part I . Roczniki PZH (Annals of
the State Institute of Hygiene) 1972, 23, 387-
2 . Illinov P . P .i V'treshni bolesti (Internal Diseases Bulgarian)
1967, 6t 367 .
*
Fiske C. H. and Subbarov J., according to J. Homolkae
Biochemical diagnosis, 3 rd edition. Palçstwowy Zaklad
Wydawnictw Lekarskich (State Medical Publishing HouSe)
Warsaw, 1961.
10. Nikonorow M. and Homrowski S.: Roczniki Wojskowego Instytütù
Higieny i Epidemiologii (Annal of the Military institute
of Hygiene and Epidemiology) 1968, 3-4, 19.
11. Rakowski W.: The cholesterol level in the adrenals of ratè
fed on a diet containing rapeseed oil. Documentation
in the Library of the Institute of the Fat Industry in
•
Warsaw, 1962.
20.
Beare-Rogers J. L.:
Proceedings
Published in Ottawa, 1970,
21.
(20-23 September 1970)
p. 450.
eare-Rogers J. L., Nera E. A. and Heggtveit H. À.
p.450
22. Kodykow B et al.: Prace instytut6w 1 Ieboratori& .Eadeirétybil
Przemyslu Spoiywczego (Works of the Institutes and
Research Laboratories of the Food Industry) 1969.,
12, 587.
25. Thomasson H. J.:
International Symposium , o4
■
•
edafiuk
Zeszyty Problemowe Postçpciw Eauk.RoZnizzych (flotes On
the Problems and Advances in Agricultural ,S:ciens) 197:0, :fasc.
91,
p. 381.
•th May, 1972
Warsaw, Powsirlska St. 63.