Blood Lipid Profile And Tissue Minerals Status Biology Essay
Abstraction: During the last two decades the sum of Zn consumed from Zn-fortified nutrients and Zn rich dietetic addendums have doubled for all age groups and the sum of this will further increase over clip. The consequence of long term increasing Zn burden in the blood lipid profile and tissue minerals position in the organic structure has non been worked out so far.
In this survey, three groups of rats were fed on semi-synthetic diet incorporating 20 mg Zn/kg ( control, group-I ) , 40mg Zn/kg ( group-II ) and 80mg Zn /kg ( group-III ) diet severally for a period of 180 yearss. The consequences revealed that the gained in organic structure weight increased in rats in Zn concentration dependant mode. The blood lipoid profile displayed a important rise in serum entire lipoids, cholesterin, triglycerides, LDL-cholesterol, VLDL- cholesterin whereas HDL-cholesterol showed a decrease in their degrees in group-II and III than their control counter parts. The tissue metal position showed an addition of Zn, Cu, Mg and Mn in the serum, a rise in Zn in liver and kidney and autumn in Cu, Mg and Mn in kidney and liver. This information suggest that extra Zn in diet when Federal for longer period of clip induced dyslipidemia and alter tissue minerals position.Cardinal Wordss: Zn, Cu, Mg, Mn, Zn- supplementation, Blood lipoid profile, Minerals position*Corresponding writer Email ID: birla.kshetri @ gmail.
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It is good documented that Zn ( Zn ) is an indispensable hint mineral which is necessary for wellness and growing and is peculiarly of import for the map and activity of over 200 metalloenzymes ( Chen et al. 1991, NRC, 1989 ) . Zinc besides acts as indirect antioxidants via stabilisation of cell membranes and the suppression of free groups production ( Bray and Better, 1990 ) . Inspite of the fact that Zn is indispensable micronutrient, there are surveies which reported that both lack and surplus of Zn can do dyslipidemia and impair hepatic cholesterin and enhance oxidative emphasis ( Subramanyam and Vijaya, 1997 ) .
Contradictory groundss and consequences were reported from different surveies sing the consequence of Zn supplementation on plasma lipoid degrees both in animate beings ( Allen and Klevay, 1978, Koo and Lee, 1989 ) and worlds ( Hopper et al.1980, Chandra, 1984, Freeland Graves et Al. 1982 ) .
The bulk of human tests surveies have shown either increase/decreased or unaffected by the supplementation of extra Zn in blood cholesterin ( AREDS Report, 2002, Bogden et Al. 1988 ) , triglycerides ( Gato and Samman, 1995 ) , LDL-cholesterol ( Chandra, 1984 ) and HDL- cholesterin degree ( Black et al. 1988 ) .Furthermore, the association of altered metal position with alteration in blood lipid profiles has been reported by different research worker otherwise. Sugawara, in 1984, found a positive correlativity between serum cholesterin and serum Cu and Zn concentration. A positive relationship between serum Zn and Zn to Cu ratio with plasma entire cholesterin and LDL-cholesterol has besides been reported ( Tully et al. 1995 ) . The ratios of Zn to Cu appear to impact the degree of lipoprotein in the blood.
High degrees of Zn consumption have been shown to take down HDL – cholesterin degrees in blood, raise entire and LDL- cholesterin, bring on thrombocyte collection and lead to coronary artery disease in animate beings ( Klevay and Hyge 1973 ) . Cu lack is associated with atherogenic alterations in lipid profiles i.e increased LDL – cholesterin and decreased HDL- cholesterin which are risk factor for cardiovascular diseases ( Klevay, 1983 ) . Furthermore, grounds is available to propose that there is association between dietetic Mg and Mn lack, plasma lipid upsets and cardiovascular diseases ( Seelig and Heggtveit, 1974, Seelig, 1980, Dorothy J. Klimis-Tavantzis et Al. 2003 ) .Even though Zn is indispensable micronutrients, the usage of Zn addendums has been discouraged by some wellness professional because inordinate consumption of Zn has been reported to bring on Cu, Mg, and Mn lacks due their counter interactions ( Aaseth 1998, Couzy et Al. 1993, Yadrik et Al.
1989 ) . This counter interaction is of practical concern because experimental informations suggested that lack these hint minerals impairs enzymes of antioxidant system and can bring forth assortment of biochemical and physiologic alterations and has been implicated in the etiology of chronic diseases ( Rayssiguier et al. 1993, Thompson et Al. 1992 ) . During the past two decennaries, there has been a rise in the ingestion of higher Zn either from Zn fortified nutrients as in the USA ( Arsenault and Brown, 2003 ) , or from veggies ( 40mg/kg or more in above land veggies and 120mg/kg above in the belowground veggies ) and meat groceries ( 120 mg Zn/kg ) as in some provinces of India ( Ram et al, 2005 ; Singh and Taneja, 2010 ) and this will increase over clip. This guided us to look into the existent state of affairs of the long term consequence of increasing Zn burden in the blood lipid profile and tissue minerals position of the organic structure.
Therefore, the supplementation of pharmacological dose of Zn in otherwise Zn equal diet was investigated with the purpose if extra Zn in the diet triggers alteration in blood lipid metamorphosis and minerals position in rats which are non genetically predisposed to any diseases. The consequences of the present survey are given in this communicating.
MATERIAL AND METHODS:
For the present probes, a semi-synthetic basal diet rich in fat and refined saccharose was preferred over the standard pellet rat diet consisting of the natural ingredients to maintain the consistence of the composing of diet peculiarly of fat, saccharose and micronutrients through out of the experiment and to govern out the possibility of Zn- interaction with fibres and phytates ( Oberleas and Harland, 1981, Turnland et al. 1984 ) which are known to cut down bioavailability of Zn by adhering it in digestive piece of land and may take longer clip continuance of feeding to attest the impact inordinate Zn in diet. Consequently, isocaloric semi-synthetic basal diet for the rats was prepared following modified Orgebin – crist et Al. 1971. It contained ( g/100g of diet ) :
Table-1: Composition of Basal Diet
*Vitamin Mixture ( mg/ kilogram )
**Mineral Mixture ( g/kg )
Casein30Ascorbic acid – 500CaH2PO4 – 25.3Agar2.
0Biotin – 4CoCl3 – 0.04Corn oil5Calcium- D- pentothenate – 320CuCl2 – 0.10Cellulose8Choline chloride – 2500FeSO4 7H2O – 0.60Sucrose51.0Folic acid – 10Mn SO4.
5H2 O – 0.31*Vitamin mixture0.50Inositol – 1000Mg SO4A .H2O – 4.
05**Mineral mixture3.50Retinol – 0.31NaF – 0.088Entire diet100Pyridoxine HCl – 80Kl – 0.004Riboflavin -120Na2CO3 -1.15Ergacalciferol – 0.
0031KCL – 3.430Thiamin HCl – 200Zn SO4.7H2O – 0.088I±-Tocopherol ethanoate ( E ) – 60Cyanocobalamin – 0.40Nicotinic acid – 300The basal diet was further divided into 2 parts: control diet consisting of basal diet incorporating 20 mg Zn/kg diet ( per-se ) for Group- I and Zn-supplement-diet-I incorporating 40 mg Zn/kg diet for Group – II and Zn supplemented-diet-II incorporating 80mg Zn/kg diet for Group-III by consequently increasing ZnSO4.7H2O in basal diet. For each diet, the mineral & A ; water-soluble vitamins were land in sucrose and fat-soluble vitamins were dissolved in maize oil. Agar, which served as a binder and was dissolved in 25 milliliter of three-base hit distilled, deionised warm H2O ( 60A°C ) .
On chilling to 40A°C, the contents of each diet were exhaustively assorted in agar solution in separate containers. The dough so formed was put in petridishes and solidified in icebox. The coagulated diet was cut into little pieces of 2 A- 2 A- 2 centimeter size and stored in the container at the temperature & gt ; – 4A°C.
Male wistar rats ( 30 ) , aged 6 hebdomads, weighing 60-70g was procured from Central Animal House, Panjab University, Chandigarh.
They were maintained in plastic coops with unstained steel top grill at room temperature ( 25A° – 28A°C ) with 10:14 hours L: D rhythms of 70-80 % RH as per usher lines of Institutional Animals Ethics Committee. They were fed on standard pellet rat provender for one hebdomad to acclimatise. Thereafter, the rats were divided into 3 groups – I, II and II and in such a manner that their average initial organic structure weights remained about similar in each group.The animate beings were fed on their several diets ad libitium and three-base hit distilled deionised H2O was made freely available to them for 180 yearss. The organic structure weights were recorded at the beginning of the dietetic intervention and thenceforth every hebdomad. After the terminal of the dietetic intervention of 180 yearss, the male rats of each group were sacrificed utilizing diethyl ether as anesthesia.
The blood samples were collected by puncturing the bosom and blood serum was prepared by centrifugating blood at 2500 revolutions per minute for 15 proceedingss. The newly prepared serum was analyzed for cholesterin ( Roeschlau et al, 1974, Allain et Al. 1974 ) , triglycerides ( McGowan et al. 1983, Fossati and Prencipe, 1982 ) , HDL-cholesterol ( Burstein et al. 1970 ) ( all by utilizing commercially available kits- Reckon Diagnostic Pvt, Ltd, Baroda, India and ERBA diagnostic Mannheim GmbH, Mannheim, Germany, supplied through Transasia Bio- Medicals LTD, Daman ) and entire lipoids ( Frings and Dunn, 1951 ) . The LDL and VLDL-cholesterol was calculated by Friedewald ‘s equation ( Friedewald et al. 1972 ) .
Zn, Cu, Mg and Mn were estimated on atomic soaking up spectrophotometer ( Electronic Corporation of India Limited, Hyderabad- AAS 4139 ) utilizing hollow cathode lamps ( 213.9nm, 324.8nm, 285.2nm and 279.5nm for Zn, Cu, Mg and Mn severally ) . Samples of serum, liver and kidney are digested individually in 3:1 v/v azotic acids and perchloric acids on a sand bath until a white ash formed. The ash was dissolved in 6 milliliter of 10mM HNO3 and filtered through ash free filter paper before analysis. Standards of Zn, Cu, Mg and Mn from Sigma Chemical Co.
, USA were prepared by dilution in three-base hit distilled deionised H2O ( TDW ) . The consequences were subjected to statistical analysis using one manner ANOVA.
Consequences and Discussion:
The information of the survey revealed that the organic structure weight of the rats increased with addition in Zn concentration in diet during the first 150 yearss of dietetic intervention in the three groups of rats and thereafter it fell in group-II and III during the following 30 yearss with regard to their weight at 150 yearss than their control opposite numbers at twenty-four hours 180 of the experiment. The information revealed that Zn is extremely powerful food which ab initio promotes addition in organic structure weight in concentration dependent mode and its prolong supplementation consequences in decrease of organic structure weight ( Table-2 ) . This is in conformance with the studies of the old surveies ( Pomp et al. 1996, Chen et al. 1996, Taneja and Mandal, 2006 ) wherein Zn more than 20mg/kg diet ( control ) and non transcending 100mg/kg diet fed to rats for a period of widening 4 to 6 hebdomads ensuing in a important addition in organic structure weight than the control has been recorded.
Investigatory surveies on research lab animate beings besides provides groundss that the anabolic effects of Zn consequences from the higher soaking up of foods i.e aminic acids ( Moran and Lyerly, 1985 ) , fatty acids ( Koo and Turk, 1977 ) and glucose ( Taneja and Arya, 1992 ) in add-on to activation protein and nucleic acid syntheses. A positive correlativity between hair Zn and organic structure weight or BMI has been seen to be ( Taneja et al.1996, Taneja and Mahajan. 1999 ) .
Table-2: Month wise organic structure weight of male rats of Group-I [ ( fed on basal diet ( Control ) ] , Group-II ( fed on Zn- supplemented- diet- I ) and Group- III ( fed on Zn supplemented diet- II ) during 180 yearss of dietetic intervention.
[ Valuess are average A± SE of 10 observation each ] .
Time continuance ( in yearss )
Group-I ( Control )
068.0 A± 0.6867.0 A± 0.
6767.2 A± 0.7930162.15 A± 1.12192.
25 A± 1.13a231.46 A± 1.67 a60199.
17 A± 1.53271.67 A± 1.
67 a297.25 A± 1.14 a90261.
36 A± 1.97318.33 A± 1.70 a363.
43 A± 0.860 a120280.50 A± 1.
70346.14 A± 1.60 a384.55 A± 1.
39 a150320.67 A± 1.13359.57 A± 1.78 a411.65 A± 1.67 a180371.
35 A± 0.82331.83 A± 1.10 a371.67 A± 1.67 aUnit of measurements: gm ; P values: a & lt ; 0.
001 ( values of group- II and group-III were compared with group-I ) .
Table-3: Blood profile of male rats of Group-I [ ( fed on basal diet ( Control ) ] , Group-II ( fed on Zn- supplemented- diet- I ) and Group- III ( fed on Zn supplemented diet- II ) during 180 yearss of dietetic intervention. [ Valuess are average A± SE of 6 observation each ] .
Group-I ( Control )
Entire Lipids*202.50 A±2.14248.17 A± 1.78 a310.33 A± 2.
86 aCholesterol*61.67 A± 2.1782.0 A± 1.
02 a115.0 A± 1.86 aTriglycerides*63.67 A± 1.56104.83 A± 1.76 a121.
67 A± 2.67 aVLDL- Cholesterol*12.7 A± 0.32020.97 A± 0.360 a24.3 A± 0.340 aHDL-Cholesterol*20.
67 A± 1.1212.67 A± 0.710 a8.83 A± 0.410 aLDL- Cholesterol*28.
27 A± 1.3548.37 A± 0.760 a81.83 A± 1.94 aUnit of measurements: * : mg/dl ; P values: a & lt ; 0.
001 ( values of group- II and group-III were compared with group-I ) .The blood profile of treated rats revealed important rise of entire lipoids, cholesterin, triglycerides, VLDL – cholesterin and LDL- cholesterin and decreased HDL-cholesterol after 180 yearss of Zn supplementation in group-II and group-III as compared to the control group-I ( Table-3 ) . These observations are consistent with old studies from both in homo and animate being surveies where supplementation of Zn increased blood cholesterin ( both LDL and VLDL-c ) and decreased in HDL- cholesterin degree ( Subramanyam and Vijaya, 1997, Chandra, 1984, Black et Al, 1988, Hopper et Al. 1980 ) . This change in serum lipoproteins profiles in the Zn treated group-II and III in the present probes may be resulted from lack of Cu, Mg and Mn in the tissues as a effect of high Zn supplementation. The appraisal of the metal position in the present survey showed that Cu, Mg and Mn were about up to 30 % less in group-II and up to 50 % less in group- III rats both in liver and kidney bespeaking the initiation of the Cu, Mg and Mn lack or rise in ratio of Zn to Cu/ Mg/Mn in the tissues.
This rise of serum entire lipoids, cholesterin, triglycerides, VLDL and LDL- cholesterin an autumn in HDL- cholesterin after 180 yearss of intervention in group-II and III rats coincide really good with grade of autumn of Cu concentration in these tissue, inspite of the fact that blood serum has higher Cu concentration than the control rat. Cu lack induced hypercholesteremia has been demonstrated in animate beings and worlds ( Bureau et al. 1998, Nielsen and Milne, 1992 ) .
The plasma cholesterin addition by increasing dietetic Zn at all degrees Cu suggests that dietetic Zn seems to stamp down the plasma cholesterin take downing consequence of dietetic Cu ( Klevay, 1975, Lata and Mehta, 1989 ) . Cholesterol appeared to unclutter from the liver to blood plasma faster in the Cu deficient rats than the control ( Allen et al. 1982, Shao and Lei, 1980 ) , as a consequence of the increased in the activity of liver enzyme, 3-hydroxy-3 methylglutamate coenzyme A reductase ( Yount et al. 1990, Koo et Al. 1993 ) , the rate restricting enzyme in cholesterin biogenesis and lessening in the lipoprotein lipase, hepatic lipase and lecithin cholesterin acyltransferase ( ICAT ) activity ( Lau and Klevay, 1981 ) .Experimental Mg lack has besides been shown to increase triglycerides, cholesterin, VLDL, LDL rich lipoproteins and decreased HDL-cholesterol ( Rayssiguier and Gueux, 1986, Gueux et Al. 1991 ) , about similar to Cu lack. In Mg deficient animate beings, hypertriglyceridemia could hold arisen as a effect of increased synthesis of triglycerides in the liver, decreased remotion of lipoid from the blood or combination of both.
An increased in the HDL-cholesterol concentration is closely associated with triglyceride metamorphosis and faulty triglyceride remotion from the plasma compartment might explicate low HDL-cholesterol degree as found in the present probes. Kawano et Al. 1987 besides observed decreased HDL- cholesterin, HDL protein and HDL apo- Tocopherol degrees with manganese lack. Because HDL atoms carry a major part of plasma cholesterin in the rat, these alterations could impact cholesterin metamorphosis in Zn treated groups. One more possible mechanism for the change of lipid degrees in manganese-deficient animate beings is that manganese lack resulted in increased lipid peroxidation of the endoplasmic Reticulum, which is the site of lipoprotein synthesis. Bell and Hurley, 1973, observed swollen and irregular endoplasmic Reticulum in tissues of Mn- deficient mice. This suggests that the ascertained rise of in degree of cholesterin, triglycerides, VLDL-c and LDL-c and decrease in HDL- cholesterin in the group-II and III rats in the present probes resulted due to Cu lack twosome with Mg and Mn lack in tissues caused by inordinate Zn in diet. Such lipid metamorphosis upsets have become common and of import jobs in clinical medical specialty.
Interest in lipid upset arises from their close relationship to vascular diseases, a major job of western and presently in developing states. Prospective informations have shown that the hazard of coronary bosom diseases is related to the serum cholesterin and triglyceride degrees ( Morrison and John, 2009, Larsa and Bottiger, 2003 ) . The part of the serum entire cholesterin to hazard has besides been found to be determined by its partitioned in the assorted lipoprotein fractions. A comparatively big sum of cholesterin in the LDL- lipoprotein cabal is atherogenic whereas that in the HDL fractions appears protective ( Gordon et al. 1977 ) .The consequences of the blood lipid profile in the present survey were consistent with impairment of the metal position in the group -II and III rats which in bend led to an addition of the cholesterin, triglycerides, VLDL-c, LDL-c and lessening in HDL-c in their several groups. The element analysis in the three groups of rats revealed that serum Zn, Cu, Mg and Mn increased significantly in group- II and III compared to those control rats in group-I.
Furthermore, Zn in liver and kidney was found to be significantly higher whereas Cu, Mg and Mn were significantly lowered in group-II and III compared to those control opposite number ( Table 4 ) . The addition of these elements were more in serum of group-III than in group-II inspite of the fact that merely the sum of Zn on the diet has been increased while Cu, Mg and Mn were equal and were in equal sum to the diet of group-I. The metal position in the blood plasma at a given clip represents a sum of edge, exchangeable and katabolic constituents of metals and hence could take to confounding consequences and anticipations.
The concentration of metal in tissues are better index of their position in the organic structure than the blood plasma in measuring metal lacks or extra since their degree are maintained by a dynamic equilibrium between tissue metals and exchangeable metal constituents of blood plasma The present informations of micro and macro elements indicated that Zn when in surplus, even in pharmacological doses in diet, over a period of clip, replaces Cu, Mg, Mn taking to their leaching in the blood and elimination in the urine even if these metals are equal in diet. This leaching of Cu, Mg and Mn from the soft tissues to the blood ensue their lacks in the tissue as observed in liver and kidney of group-II and III rats.
Table-4: Average Zinc ( Zn ) , Copper ( Cu ) , Magnesium ( Mg ) and Manganese ( Mn ) concentration in the serum, liver and kidney of male rats of Group-I [ ( fed on basal diet ( Control ) ] , Group-II ( fed on Zn- supplemented- diet- I ) and Group- III ( fed on Zn supplemented diet- II ) during 180 yearss of dietetic intervention. [ Valuess are average A± SE of 10 observation each ] .
Group-I ( Control )
Serum Zn*0.57 A± 0.051.70 A± 0.
02 a1.90 A± 0.05 aSerum Cu*0.95 A± 0.042.35 A± 0.
05 a3.23 A± 0.14 aSerum Mg*1.56 A± 0,081.70 A± 0.06 a1.96 A± 0.
04 aSerum Mn*0.21 A± 0.050.67 A± 0.05 a1.
01 A± 0.06 aLiver Zn @32.7 A± 0.8844.
1 A± 0.84 a57.0 A± 0.65 aLiver Cu @60.3 A± 0.6843.
9 A± 1.13 a30.3 A± 0.77 aLiver Mg @61.0 A± 1.
0348.0 A± 0.49 a34.0 A± 0.
90 aLiver Mn @47.80 A± 0.5936.40 A± 0.65 a30.90 A± 0.66 aKidney Zn @35.4 A± 0.
5442.7 A± 0.69 a55.5 A± 1.52 aKidney Cu @53.5 A± 0.7446.50 A± 0.
68 a38.10 A± 0.63 aKidney Mg @54.50 A± 0.8445.
90 A± 0.54 a35.20 A± 0.76 aKidney Mn @42.80 A± 1.8737.
90 A± 0.94 a28.20 A± 0.62 aUnit of measurements: * : mg/dl ; @ : Aµg/g tissue weight ; P values: a & lt ; 0.
001 ( values of group- II and group-III were compared with group-I ) .However, in the present survey urinary Zn, Cu, Mg and Mn degree was non assessed because merely hint sums are found in the piss and urinary loss of metal is merely little fraction of their turnover, thereby restricting its usage as a sensitive index of metal position. The ascertained addition of Zn degree in liver and kidney in group -II and III rats than their control group-I may be attributed to the overexpression of Zn metallotheionein cistron during intervention period due to high and prolong Zn consumption. As a consequence of this they absorbed and retained greater sum of Zn than their control opposite number taking to lacks of Cu, Mg and Mn due to their interactions ( Irato et al. 1996 ; Nielsen and Milne, 2004 ) .The consequences of the present survey therefore supply strong grounds that inordinate Zn in diet even in pharmacological doses alter blood lipid metamorphosis and deteriorated mineral position of the organic structure.
These findings are peculiarly important for the people of some States of India where the groceries are overloaded with Zn ( Singh and Taneja, 2010, Ram et Al. 2005 ) . The lifting prevalence of cardiovascular related diseases even in the immature age population during the past two decennaries may non basically due to alter in life style but besides may be associated with inordinate consumption of Zn from Zn rich nutrients which in bend may change hint minerals and lipoproteins metamorphosis of the organic structure up to a degree plenty to attest as diseases. Therefore, there look to be several possible inauspicious effects of such pharmacological doses of Zn peculiarly when such consumptions are continued for a drawn-out period.
Thankss are due Prof.
S. Chaudhary, former Chairman, Department of Zoology, Panjab University, Chandigarh, for supplying the necessary research lab installations. Financial aid to K. Birla Singh under support strategies of Minor Research Project by University Grant Commission-North Eastern Regional Office ( UGC-NERO ) , Guwahati, is appreciatively acknowledged.