Compared levels
of specific divalent trace elements in hyperlipidemia
and hepatitis sera patients
......................................................................................................................................................................
Namama Soran
H (1)
Dler. M. Salh (1)
Avan Arif A (2)
(1) Chemistry Department,
School of Science, Faculty of Science and Educational
Science,
Sulaimaniyah University, Kurdistan Region, Iraq.
(2) Ministry of Health. Central Lab. Sulaimani,
Iraq
Correspondence:
Email: sareaali@hotmail.com,
dlerchem@yahoo.com
ABSTRACT
Patients with hepatitis infection can also
have hyperlipidemia. Sera of both hepatitis
(40) and hyperlipidemia (40) were collected
from the central lab of Sulaimaniyah, Kurdistan
region, Iraq. Age, sex, duration of infection,
medication used and lipid profile using
computerized database program, as characteristics
information of patients were done. The levels
of Cr, Cu, Zn, Mn, V and Se were determined
in sera of both patients and compared with
healthy controls (n=30), using Inductively
Coupled Plasma-Optical Emission Spectrometry
(ICP-OES). Hepatitis samples yielded concentration
in ppb of (7.34±0.01) Cr, (171.7±2.51)
Cu, (138±3.41) Zn, (1.309±0.99)
Mn, (184.7±6.12) V and (218.9±3.14)
Se respectively, while hyperlipidemia samples
yielded concentration (ppb) of (5.22±0.45)
Cr, (197.01±2.34) Cu, (160.3±4.01)
Zn, (0.068±1.91) Mn, (247.01±0.65)
V and (185.5±3.11) Se respectively.
Control samples reflect the following concentrations
(ppm), (5.324±1.34) Cr, (131.2±4.34)
Cu, (216.6±0.83) Zn, (0.024±1.75)
Mn, (235.3±0.18) V and (223.2±3.15)
Se respectively. The aim of the study focused
on evaluating metal ions physiological role
in the body, and as liver was responsible
for synthesizing lipid, thus lipid profiles
were suggested to be investigated along
the study.
Key words: hepatitis, hyperlipidemia,
trace elements, ICP-OES technique
|
Hepatitis and hyperlipidemia
have become a major human health problem worldwide
[1]. It is not clear what the prevalence of hyperlipidemia
is and how often hyperlipidemia is treated in
patients with hepatitis. In addition, in those
patients receiving cholesterol-lowering medication,
it is not clear whether it is associated with
worsening of liver synthetic function or not.
In 1978, a comprehensive compilation of literature
values was published by Iyengar et al. [2]. It
served as a guideline for approximate concentrations
of many elements in numerous tissues. In 1983,
Woittiez [3] exclusively investigated the problem
of establishing reference values for 28 elements
in human serum. Trace elements such as Cr, Cu,
Zn, Mn, V and Se are essential nutrients for humans
and are required in very small amounts for many
physiological functions, including immune, antioxidant
function, growth and reproduction [4]. It has
been determined that humans need nearly 72 trace
elements, including very low concentrations of
heavy metals, such as Cu, Se, V, Cr, Mo, Mn and
Co. Most metals are toxics at high concentrations,
while others provoke deleterious effects at low
concentrations [5]. For example, Vanadium (VI)
revealed a biological interest due to its biotoxicity
[9]. The significance of the biochemical and nutritional
roles of trace elements is widely recognized,
since metals are found as constituent components
of many metalloproteins and metalloenzymes. Some
trace elements such as Copper act as cofactors
against hepatic fibrosis in chronic liver diseases.
Trace elements also affect many aspects of lipids
metabolism through enzymes action and have modulator
effects on the synthesis and metabolism of lipids
[10]. Zinc for example, functions as an antioxidant
and stabilizes membranes; Selenium is an essential
micronutrient for human health [11-13]. The human
body contains approximately ten milligrams of
Mg most of which is found in the liver, bone and
kidneys; it is a cofactor for a number of important
enzymes, including arginase, pyruvate carboxilase
and several phosphatase, peptidases and glycosyl
transferase. Low levels of Mg have been associated
with Atherosclerosis [14]. We conducted a study
to assess the relationship and changing of some
trace elements between hyperlipidemia and hepatitis.
The randomly selected study
group comprised 40 patients with hepatitis that
included 25 males and 15 females (aged 30±15.2),
ranging between 25 and 60 years. Forty patients
with hyperlipidemia were also included, 23 males
and 17 females aged 35±12.3 years. The
control group comprised 30 healthy individuals
, which included 19 males and 11 females aged
between 20 and 63 years. Sera of patients and
controls were isolated from blood at the central
lab (Sulaimaniyah- Kurdistan region, Iraq). All
sera were collected in the morning after fasting
8 hours. Patients with hepatitis were diagnosed
based on clinical, biochemical and histological
data. Serum with hyperlipidemia was also diagnosed
based on increased concentration of cholesterol,
triglyceride, HDL and LDL. Standard solution of
the metals, 1000 µg/ml of Cr, Cu, Mn, Zn,
V and Se, were prepared. Other chemicals were
purchased from Fluka. Standard solutions were
prepared freshly from the stocks, with diluted
nitric acid (3 %v/v). In order to achieve ICP-OES
responses, the experiments were performed using
different concentration levels.
Sample Digesion:
1ml of serum was transferred to a Teflon beaker
and 10ml of concentrated nitric acid and 2.5ml
concentration perchloric acid were added. The
sample was then brought very slowly to boiling
on a hot plate and heated to dryness. If sample
blackening occurred during the fuming stage, nitric
acid was added drop wise, then the sample was
cooled, dissolved again in distilled water and
concentrated HCl (10:1) and brought to a volume
of 25ml in a volumetric flask. The solution was
analyzed against calibration curve [16].
ICP-OES:
An inductively coupled plasma-Optical Emission
Spectrometer has been extensively used in the
analysis of major, minor and trace elements in
biological material because of its high sensitivity,
accuracy, low matrix effect and simpler operation.
The presence of various elements in the sample
was identified by determining the wavelength of
the emitted radiation (Cu: 327.393nm, Se: 196.026nm,
Zn: 213.857nm, Cr: 267.716, Mn: 257.610, V: 290.880)
and the concentration was calculated by intensity
of the radiation, which might be sufficiently
low for certain applications with a simple matrix.
Sample and standard were analyzed in triplicate
[17]. Statistical analysis, using STATISTICAL
program (statsoft) was applied for data analysis.
A p-value of < 0.05 was considered statistically
significant [18].
Serum concentrations of total
cholesterol, HDL, LDL and triglyceride are represented
in Table 1. Patients with hyperlipidemia show
a significant decrease (p<0.05) in Zn serum
(160.3±4.01), (0.068±1.91) Mn and
Se (185.5±3.11) ppb respectively compared
with controls, while levels of Cu and V increased
significantly (p<0.05) compared with those
of controls. No significant changes were found
in case of Cr.
Table 1 : Characteristics of both patients which
show the same features
Serum Cr, Cu, and Mn of hepatitis patients were
significantly higher (17.53±0.01), (171.7±2.51)
and (1.309±0.99) ppb respectively compared
to normal (Cr=5.324±1.34), (Cu=131.2±5.93)
and (Mn=0.204±1.75) ppb. The serum Zn,
V and Se level was (Zn=138.3±3.41), (V=184.7±6,
12) and (Se=218.9±3.41) ppb in patients
with hepatitis which was higher than normal, (216.6±0.83),
(235.3±0.18) and (223.2±3.15) respectively,
as shown in Table 2. Results of this study have
been summarized and show the overall comparison
between trace elements in both cases of hyperlipidemia
and hepatitis in Figure 1.
Table 2: Concentrations of Cr, Cu, Zn, Mn,
V and Se in healthy control hepatitis and hyperlipidemic
patients
Figure 1
Damage of hepato cells can result
in fluctuation of body cell constituents. Lipid
profiles are of importance and can be affected
by liver damage and the process results in variation
in lipid levels. Trace elements are used as a
diagnosing tool during disease; it is important
to know whether the balance is changed in free
or bound elements. The results of the present
study have shown a significant increase (p<
0.05) in Cr, Cu and Mn in hepatitis and Cu and
V in hyperlipidemia, while there were no significant
changes in serum Cr in hyperlipidemia patients.
These metals are members of one of the major subgroups
of the micronutrients that have attained prominence
in human nutrition and health. The biological
role of trace metals, especially serum Zn, Cu,
Cr and Mn, in different physiologic conditions
has been extensively investigated in recent years
(19). Similar observations were made by Lin CC
et al. (20), and Pramoolsinsap C (21). They reported
statistically significant decreased levels of
serum Mn, Cu and Se in patients with hepatitis.
But different observations have been reported
by Saghir M. et al. (22), and show that Cu level
decreased in hepatitis. It is clear that deficiencies
of some trace elements, such as Cu, Cr, Zn and
Mn can result in marked alterations in lipid and
lipoprotein metabolism (23). To the best of our
knowledge, there has been no previous research
regarding the correlations of serum trace elements
with lipids and lipoprotein in hyperlipidemia
patients. Several studies reported an inverse
relation between serum Cu and cholesterol in rats
during Cu deficiency (4), while Koo and Williams
found no significant correlation between the serum
Cu and cholesterol levels in non Cu deficient
rats (11). Decrease in serum Se might indicate
the development and progression of hepatitis,
it also links to the disease progress of some
viral agents in relation to the biosynthesis of
selenoproteins (24), and decrease in serum Se
significantly increases the risk of cancer mortality.
Four-year animal studies showed that dietary supplement
of Se reduced the hepatitis infection by 77.2%
(25). El-Hendy et al, showed that Zn deficiency
increases serum cholesterol in a dose-dependent
rat (26). Manganese is critical for lipid and
lipoprotein metabolism; it has been demonstrated
that Mn enhances cholesterol synthesis in the
liver. The above results show that serum Cu concentrations
of hepatitis patients are higher than normal individual
serum concentrations. These elevated serum Cu
levels indicate an alteration of Cu metabolism
during the acute phase of uncomplicated hepatitis
(27). It may be explained by the release of copper
from damaged necrotic hepatocytes (28).
Vanadium has a role in the regulation of the metabolism
of lipids and other constituents of importance.
The major concern is that excessive levels of
vanadium have been suggested to be a factor in
manic depression, as increased levels of vanadium
is found in hair samples from manic patients,
and these values fall towards normal levels with
recovery (29).
The real mechanism is not known
but abnormal results of trace elements may damage
the liver by oxidative stress. Our study results
suggest trace element supplementation may be complementary
therapy to hyperlipidemia and hepatitis patients,
so some of these trace elements might be considered
as a marker of normal liver function. Dietary
intake of these elements or vegetable and food
which are considered as rich sources of these
elements are necessary to reduce these two syndromes.
1) Sankaraya, S, Oncu, S. Ozturk, B. Effect of preventive
applications on prevalence of hepatitis B virus
and hepatitis C virus infections in west Turkey.
Saudi Med.J. 2004; 25(8), 1070-82.
2) Iyengar GV, Kollmer WE, Bowen HJM. Elemental
composition of human tissues and body fluids. Weinheim:Verlag
Chemis,1978.
3) Woittiez JRW. Elemental analysis of human serum
and serum protein fractions by thermal neutron activation
(phd thesis). Amsterdam: University of Amsterdam,1983.
4) Cunnane, S,C. Role of zinc in lipid and fatty
acid metabolism. Prog.food.Nutr.Sci.1988; 12, 151-188.
5) Rizk, SL; Sky-PECK, H,H. Comparison between concentrations
of trace elements in normal and neoplastic human
breast tissue. Cancer Res. 1984; 44; 5390-5394.
6) Kammerer, M., Mastain, O.; LE Drean-Ouenech,
DU, S.; Pouliquen,H.; Larhantec, M. Liver and kidney
concentrations of vanadium in oiled seabirds after
the Erika Wreck. Scien. Total Environ. 2004; 333(1-3):295-302.
7) Bin, QH; Garfinkel, D. The cadmium toxicity hypothesis
of aging: a possible explanation for the zinc deficiency
hypothesis of aging. Med. Hypothesis. 1994; 42;
380-384.
8) Dowd, T.L; Rosen, J.F; Gundberg, C.M; Gupta,
R.K. The displacement of calcium from osteocalcin
at submicromolar concentrations of free lead. Biochem.
Biophys. Acta. 1994; 122(6):131-137.
9) Rojas, E.; Valverde, M.; Herrera, L.A.; Altamirano-Lozano,
M.; Ostrosky-Wegman, P. Genotoxicity of Vanadium
pentoxide evaluate by the single cell gel electrophoresis
assay in human lymphocytes. Mutat.Res. 1996, 359;
77-84.
10) Suzuki K, Oyama R, Hayashi E, Arakawa Y. Liver
disease and essential trace elements. (Article in
Japanese), Nihon Rinsho. 1996 Jan; 54(1):85-92.
11) Koo, SI, Williams D.A. Relationship between
the nutritional status of zinc and cholesterol concentration
of serum lipoprotein in adult male rats. Am. J.
Clin. Nutr. 1981, 34, 2376-2381.
12) Shankar AH, Prasad AS. Zinc and altered resistance
to infection. Am J. clin Nutr. 1998, 68; 447S-463S.
13) Rayman M.P. The importance of Selenium to human
health. Lancet. 2000, 356, 233-34.
14) Blaurock-Busch E. Mineral and trace element
analysis, Laboratory and clinical application. Tmi
1997.
15) Raghunath, R.; R.M. Tripathi, K. Vinod, A.P.
Sathe, R.N. Khandekar, K.S.V. Nambi. Assessment
of pb, Cd, Cu and Zn exposures of 6-19 year old
children in Mumbai, Environ. Res. 1991, 80, 215-222.
16) Nakayama, A.H., Fukuda, M. Ebara, H. Hamasaki,
K. Nakajima, H. Skural, A. New diagnostic method
for chronic hepatitis, liver cirrhosis and hepatocellular
carcinoma based on serum metallothionein copper
and zinc levels. Bioliphar. Bull. 2002; 25(4), 426-431.
17) R. Selvaraju, R. Ganapathi Raman, R. Narayana
Swamy, R. Vallappan, R. Baskaran. Trace element
analysis in hepatitis B affected human blood serum
by inductively coupled plasma-Atomic Emission spectroscopy
(ICP-AES). ROMANIAN, J. BIOPHYS. 2009; 19(1):35-42.
18) Joaquim P. Applied statistics using SPSS, statistica,
Matlab and R. 2nd ed., Springer company USA. 2007;
205-211,451-499.
19) Hambidge M. Human zinc deficiency. J. Nutr.
2000; 130, 1344-9.
20) Lin CC, Huang JF, Tsai LY, Huang YL. Selenium,
iron, copper and zinc levels and copper-to zinc
ratios in serum of patients at different stages
of viral hepatic diseases. Biol Trace Elem Res.
2006, 109(1):15-24.
21) Pramoolsinsap C, Promvanit N, Kurathong S. Serum
trace metal levels in patients with acute hepatitis
B. Southeast Asian J Trop Med public health. 1996,
27(3):476-80.
22) Saghir M, Shaheen N, Shah MH. Comparative evaluation
of trace metals in the blood of hepatitis C patients
and healthy donors. Biol trace elem res. 2011, 143(2):751-63.
23) Engle T.E, Spears J.W., Xi L. Concentration
in fishing stress dietary copper effects on lipid
metabolism and circulating catecholamine. J. Am.Sci.
2000,78:2737-2744.
24) Banares F.F.,E. Cabre, M. Esteve. Serum Se and
risk of large size colorectal adenomas in a geographical
area with a low Se status. Am. J. Gastroenterol.
2002,97:2103-8.
25) Yu. S.Y., Y.J. Zhu, W.G. Li. Protective role
of Se against hepatitis B virus and primary liver
cancer in Qidong. Biol. Trace Elem. Res. 1997, 56(1):117-124.
26) El-Hendy H.A., Yousef M.I., Abo El-Naga N.I.
Effect of dietary zinc deficiency on hematological
and biochemical parameters and concentration of
zinc, copper and iron in growing rats. Toxicol.
2001,167:163-170.
27) Hatano R. Accumulation of copper in liver and
hepatic injury in chronic hepatitis C. J. Gastroenterol
Hepatol. 2000,15:786-779.
28) Taylor E.W., R.G. Nadimpalli, C.S. Ramanathen.
Genomic structures of viral agents in relation to
biosynthesis of selenproteins. Biol Trace Elem.
Res. 1997, 56: 63-91.
29) Goujy, Han CC, Lin YM. A contemporary treatment
approach to both Diabetes and Depression by cordyceps
sinensis, rich in Vanadium. Evid Based complement
Alternat Med. 2010,7(3):387-9.
|