Pharmacologica/ Research Communications, Vol. 9, No. 9, 1977
MODIFICATIONS OF LIPIDS, LIPOPROTEINSAND PLASMATICAPOLIPOPROTEINS INDUCED BY A PHOSPHOLIPID EXTRACTEDFROMCALF HEART. Avogaro P., Cazzolato G., Bittolo Bon G., Quinci G.B., Belussi F., Hantero O. e Conti F. Regional General Hospital, Venice, Italy Unit for Atherosclerosis, Hyperlipemias a. Diabetes. Ospedale"Ca Granda" Divisione Vergani, Milano J
Receivedin final form 24 September 1977
ABSTRACT 11 hyperlipemic subjects were given intravenously 200 mg daily of purified phospholipids, iisolated from calf heart, for 20 days. A significant decreaseiof total cholesterol, LDLJ cholesterol, apo-B-LDL Land apo-AI-VLDLwas observed. Levels of total apo-B and apo-AI-HDL were also lowered even not signlficantly. I
INTRODUCTION
The recent conclusions of the coronary Drug Project (The Coronary Drug Project Research Group, 19~5) have emphasized, that none of the more widely used antili/pemic drugs, show, any •
(
effectiveness in reducing cardiovascular morbility or mortality rate. Studies on drugs, which might reduce the effects of athero sclerosis, are therefore of interest. A regression of athero
885
Pharmacological Research Communications, Vol. 9, No. 9, 1977
886
sclerosis has been recently described (Stafford a. Day, 1975), following the administration of naturally occuring phospholipids. This communication refers the effects of the intravenousjadministration of an extractive phospholipidic /
/
complex, named Cellucor (xx) on plasma lipoproteins and apo lipoproteins in man. The results indicated that this compound: a) reduces the levels of plasmatic lipids in the studied subjects b) moreover, modification on lipoprotein structures and of apolipoproteins is found after administration pf Cellucor.
PATIENTS
I I hyperlipemic patients were examined. The classifi cation was done, following the indications of the World Health Organization (Memorandum W.H.O., 1970). Seven patients appeared to be affected by type IIA hyperlipemia and 2 patients by the f
type IIB. Other 2 patients, to whom a type IIA hYperlipemia was o r i g i n a l l y attributed, showed normal lipemia values at the beginning of the experiments, as a consequence of a long term anti-atherogenic d i e t ( P a t r a s s i , Avogaro a. Crepaldi, 1967).
(xx) The composition of lO mg of phospholipids is: phosphatidycoline + lisophosphatidylcoline + sulphatides . . . . . . . . . . . . . . . .
= 51.8%
Sphingomyelin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
= If.O%
Phosphatidylethanolamine + Phosphatidylserine . . . . . . . . . = 14.3% Cardiolipin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
= 4.1%
Plasmalogens + Phosphatidylinositol + phosphatidic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
= 18.8%
Pharmacological Research Communications, Vol. 9, No. 9, 1977 No change of the diet was done during the time of the experi ments.The body-weight of the patients was daily controlled; variations were less than + l Kg during 20 days of the experiments. METHODS
A dispersion of Cellucor in saline (200 mg/250 ml) was administered by a 2 hrs. perfusion to patients, which were previously fasted for a least 12 hours. Samples of blood were taken from all patients before the perfu sion and after 20 days of treatment; from 6 patients collection was performed also at the lOth day. Blood was immediately centrifuged in tubes cpntaining an l mg/ml EDTA solution. The obtained plasma was used for the determination of cholesterol, triglycerides and total proteins. Moreover, lipoprotein fractions with dI.063 (HDL) were separated with an ultracentrifuge Spinco-Beckman L2-B65 with f i x angle, according to the Manual of Lipid Clinics Operations (Manual of Lipid Operations, 1974). Cholesterol content of each fraction was than determined. Both apolipopro teins, "B" and "Al
were determined in total plasma and in each
lipoprotein fraction, according to the Electroimmunoassay procedure (Curry et A l . , 1976). Statistical correlation on the data was performed, taking into account all the subjects, without any reference to the different lipoproteinemic phenotypes. The techniques of "between pairs" was used for the statistical analysis (Yamane, 1973).
887
Pharmacolog~al Research Comrnun~ations, VoL 9, No. 9, 1977
888 RESULTS (Table_ss l &21
The value of total cholesterolemia shows a significant reduction (pLO.Ol). The decrease was observed in lO cases over I I , with a reduction smaller than I0% in 3 cases, greater than I0% in 2 cases and higher than 15% in 5 cases. An increase of the \ cholesterolemia was observed in I case~ The variations of the 4--
\
triglycerides were not significant despite a reduction in 5 cases over I I . In 4 patients the decrease was greater than 15%. An increase was recorded in 5 cases.- The C-VLDL and the C-HDL were not substantially modified. The C-LDL showed a Significant decrease (PC0.05) in I0 cases over I I . In 2 patients the decrease was less than 10%, but in 8 subjects was greater than 15%. In one subject was observed an increase of the C-LDL. During the treatment the total apo-B decreased in 8 subjects over II with a loweriqg of 19 mg%. The reduction however was not signi ficant. The total Apo-Al did not change. Regarding the modifi cations of the apoproteins within the different lipoproteic classes, Apo-B-VLDL did not change, whereas apo-B-LDL has decreased significantly (pLO.Ol). This reduction, in average of 40 mg%, has regarded 9 subjects over I I , with a decrease smaller than I0% in 4 and greater t~an 15% in 5. Apo-AI-HDL was reduced in an average of 16 mg%, concerning 7 subjects over I I , which was not significant.
20"
1"
,
P
MODIFICATIONS OF LIPIDS, LIPOPROTEINSAND PLASMATICAPOLIPOPROTEINS INDUCED BY A PHOSPHOLIPID EXTRACTEDFROMCALF HEART. Avogaro P., Cazzolato G., Bittolo Bon G., Quinci G.B., Belussi F., Hantero O. e Conti F. Regional General Hospital, Venice, Italy Unit for Atherosclerosis, Hyperlipemias a. Diabetes. Ospedale"Ca Granda" Divisione Vergani, Milano J
Receivedin final form 24 September 1977
ABSTRACT 11 hyperlipemic subjects were given intravenously 200 mg daily of purified phospholipids, iisolated from calf heart, for 20 days. A significant decreaseiof total cholesterol, LDLJ cholesterol, apo-B-LDL Land apo-AI-VLDLwas observed. Levels of total apo-B and apo-AI-HDL were also lowered even not signlficantly. I
INTRODUCTION
The recent conclusions of the coronary Drug Project (The Coronary Drug Project Research Group, 19~5) have emphasized, that none of the more widely used antili/pemic drugs, show, any •
(
effectiveness in reducing cardiovascular morbility or mortality rate. Studies on drugs, which might reduce the effects of athero sclerosis, are therefore of interest. A regression of athero
885
Pharmacological Research Communications, Vol. 9, No. 9, 1977
886
sclerosis has been recently described (Stafford a. Day, 1975), following the administration of naturally occuring phospholipids. This communication refers the effects of the intravenousjadministration of an extractive phospholipidic /
/
complex, named Cellucor (xx) on plasma lipoproteins and apo lipoproteins in man. The results indicated that this compound: a) reduces the levels of plasmatic lipids in the studied subjects b) moreover, modification on lipoprotein structures and of apolipoproteins is found after administration pf Cellucor.
PATIENTS
I I hyperlipemic patients were examined. The classifi cation was done, following the indications of the World Health Organization (Memorandum W.H.O., 1970). Seven patients appeared to be affected by type IIA hyperlipemia and 2 patients by the f
type IIB. Other 2 patients, to whom a type IIA hYperlipemia was o r i g i n a l l y attributed, showed normal lipemia values at the beginning of the experiments, as a consequence of a long term anti-atherogenic d i e t ( P a t r a s s i , Avogaro a. Crepaldi, 1967).
(xx) The composition of lO mg of phospholipids is: phosphatidycoline + lisophosphatidylcoline + sulphatides . . . . . . . . . . . . . . . .
= 51.8%
Sphingomyelin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
= If.O%
Phosphatidylethanolamine + Phosphatidylserine . . . . . . . . . = 14.3% Cardiolipin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
= 4.1%
Plasmalogens + Phosphatidylinositol + phosphatidic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
= 18.8%
Pharmacological Research Communications, Vol. 9, No. 9, 1977 No change of the diet was done during the time of the experi ments.The body-weight of the patients was daily controlled; variations were less than + l Kg during 20 days of the experiments. METHODS
A dispersion of Cellucor in saline (200 mg/250 ml) was administered by a 2 hrs. perfusion to patients, which were previously fasted for a least 12 hours. Samples of blood were taken from all patients before the perfu sion and after 20 days of treatment; from 6 patients collection was performed also at the lOth day. Blood was immediately centrifuged in tubes cpntaining an l mg/ml EDTA solution. The obtained plasma was used for the determination of cholesterol, triglycerides and total proteins. Moreover, lipoprotein fractions with dI.063 (HDL) were separated with an ultracentrifuge Spinco-Beckman L2-B65 with f i x angle, according to the Manual of Lipid Clinics Operations (Manual of Lipid Operations, 1974). Cholesterol content of each fraction was than determined. Both apolipopro teins, "B" and "Al
were determined in total plasma and in each
lipoprotein fraction, according to the Electroimmunoassay procedure (Curry et A l . , 1976). Statistical correlation on the data was performed, taking into account all the subjects, without any reference to the different lipoproteinemic phenotypes. The techniques of "between pairs" was used for the statistical analysis (Yamane, 1973).
887
Pharmacolog~al Research Comrnun~ations, VoL 9, No. 9, 1977
888 RESULTS (Table_ss l &21
The value of total cholesterolemia shows a significant reduction (pLO.Ol). The decrease was observed in lO cases over I I , with a reduction smaller than I0% in 3 cases, greater than I0% in 2 cases and higher than 15% in 5 cases. An increase of the \ cholesterolemia was observed in I case~ The variations of the 4--
\
triglycerides were not significant despite a reduction in 5 cases over I I . In 4 patients the decrease was greater than 15%. An increase was recorded in 5 cases.- The C-VLDL and the C-HDL were not substantially modified. The C-LDL showed a Significant decrease (PC0.05) in I0 cases over I I . In 2 patients the decrease was less than 10%, but in 8 subjects was greater than 15%. In one subject was observed an increase of the C-LDL. During the treatment the total apo-B decreased in 8 subjects over II with a loweriqg of 19 mg%. The reduction however was not signi ficant. The total Apo-Al did not change. Regarding the modifi cations of the apoproteins within the different lipoproteic classes, Apo-B-VLDL did not change, whereas apo-B-LDL has decreased significantly (pLO.Ol). This reduction, in average of 40 mg%, has regarded 9 subjects over I I , with a decrease smaller than I0% in 4 and greater t~an 15% in 5. Apo-AI-HDL was reduced in an average of 16 mg%, concerning 7 subjects over I I , which was not significant.
20"
1"
,
P
