You are on page 1of 7

1

Original Scientific Paper

Biochemistry Section, Department of Basic Medical Sciences,


Faculty of Medical Sciences
University of the West Indies, Mona
Biotechnology Centre, University of the West Indies,
Mona, Kingston 7, Jamaica
E-mail: fomoruyi@uwimona.edu.jm

HEPATIC FUNCTION ENZYMES AND LIPID PEROXIDATION IN


STREPTOZOTOCIN-INDUCED DIABETIC RATS FED BITTER
YAM (DIOSCOREA POLYGONOIDES) STEROIDAL
SAPOGENIN EXTRACT
Marie A. McAnuff1, Felix O. Omoruyi1, Errol Y. Morrison1, Helen N. Asemota1,2

Key words: lipid, transaminase, yam, sapogenin, diabetes,


streptozotocin

alterations in the lipid composition of the liver with subsequent


reduction in lipid peroxidation. Data from this study also
showed improvement in liver damage associated with diabetes.

SUMMARY
INTRODUCTION
Diabetic male wistar rats (n=32) were fed diets supplemented
with 1% bitter yam steroidal sapogenin extract or commercial
diosgenin for three weeks. Thiobarbituric acid reactive
substances (TBARS), conjugated dienes, lipid profile and the
activities of alanine and aspartate transaminases and acid
phosphatase were measured in the liver. Diabetic rats (groups
B-D) lost weight significantly (p<0.05) compared with the
normal group even though there was no significant difference
(p<0.05) in their feed intake. There was no significant change
(p<0.05) in liver weight. Test diets significantly lowered plasma
glucose level. Total cholesterol and VLDL-cholesterol decreased
significantly (p<0.05), while HDL-cholesterol levels increased
significantly in group C and D rats compared to group B rats.
There was no significant change in the levels of phospholipids
and triglycerides. The feeding of commercial diosgenin and yam
sapogenin extract resulted in a significant decrease in conjugated
dienes and TBARS compared with the diabetic group fed
normal diet and normal rats. Yam extract and commercial
diosgenin also caused a significant decrease in the diabetesinduced increase in the activity of alanine transaminase.
However, they had no effect on the aspartate and acid
phosphatase levels. The study showed that the feeding of bitter
yam steroidal sapogenin extract to diabetic rats may result in

Diabetologia Croatica 32-1, 2003

Diabetes is by far the most common of endocrine


disorders and a major threat to health care worldwide.
It is projected that by 2010, at least 239 million people
will be affected by the disease (1). Diabetes poses a
threat to developing as well as developed countries. In
America, for example, diabetes ranks sixth as the
primary cause of death, and has an estimated economic
cost of 85 to 92 billion USD, two-thirds of which are
the result of productivity losses because of admission
to hospital or death (2). The prevalence of diabetes is
high in the Caribbean; in Jamaica, for example, the
point prevalence of diabetes mellitus in the 15 age
group is estimated to be 17.9% (3).
Oxidative stress is thought to be a major contributor
to cardiovascular disease in diabetes mellitus (4). In
fact, it is well documented that diabetes is associated
with increased oxidative stress, as evidenced by the
increased accumulation of lipid peroxides in the plasma
of rats (5) and humans (6) with diabetes mellitus.
Several species of wild yams are known to be
biologically active against hypertension, arthritis,
diabetes mellitus and other physical ailments (7), and

17

M. A. McAnuff, F. O. Omoruyi, E. Y. Morrison, H. N. Asemota / HEPATIC FUNCTION ENZYMES AND LIPID PEROXIDATION IN
STREPTOZOTOCIN-INDUCED DIABETIC RATS FED BITTER YAM (DIOSCOREA POLYGONOIDES)
STEROIDAL SAPOGENIN EXTRACT

have been used in traditional medicine in Africa, among


the Chinese and other Asiatic people. The saponins or
sapogenins present may be the active principles in
these tubers (8). The Jamaican wild yam (Dioscorea
polygonoides) is used in the preparation of root tonic and
contains high levels of sapogenin/saponin. The purpose
of the present studies, therefore, was to examine the
effects of yam steroidal sapogenin extract on liver lipid
peroxidation and function enzymes in streptozotocininduced diabetic rats.
MATERIAL AND METHODS

Sample preparation
Samples of wild yam were obtained from Muirhouse,
in the Parish of St. Ann, Jamaica. The sapogenins were
extracted using the method of Morris et al. (9). The
sapogenin extract from bitter yam contains 80%
diosgenin, the remaining 20% being made up of sitosterol, pennogenin, stigmasterol and 2 diosgenin.

Animals and diets


Male wistar rats were obtained from the Animal
House, University of the West Indies. The rats were
divided into four groups of eight rats each (average
weight 248.98 g) as follows: healthy rats receiving
normal diet (group A); diabetic rats receiving normal
diet (group B); diabetic rats receiving 1% bitter yam
sapogenin extract (group C); and diabetic rats fed 1%
commercial diosgenin (Sigma Chemical Co., St. Louis,
MO, USA) (group D). The normal diet (PMI Feed Inc.
Lab diet #5001) was a commercial laboratory diet
recommended for rats, hamsters and mice with the
nutrient composition of 23% protein, 4.5% fat, 6.0%
fiber, 8.0% ash and 58.5% carbohydrate.
Groups B-E received a single injection of
streptozotocin (Sigma Aldrich, 65 mg/kg body weight
in 0.05 M citrate buffer, pH 4.5), while group A
received a single injection of an equivalent amount of
buffer. After 8 days, blood was collected from the tail
and the level of blood glucose was determined. Rats
were considered diabetic if their blood glucose level
was four times in excess of the normal.
The rats were housed in stainless steel wire cages at
room temperature with a 12-h light/dark cycle and were
allowed free access to their respective diet and water.

18

The cages were cleaned daily and total feed intake was
recorded daily. The rats were kept on their respective
diet for 21 days and killed after an overnight fast by
decapitation. The facilities met the requirements of
the Institutional Animal Care and Use Committee.
Fasting plasma and liver samples were obtained for
further analyses.
Lipid peroxidation assays. Liver lipid peroxidation was
determined using the procedure of Tappel and Zalkin
(10). Lipid peroxidation was assessed as the amount of
thiobarbituric acid reactive substances (TBARS)
produced.
Conjugate diene analysis. The liver conjugated dienes
were determined by the method of Hu et al. (11). Liver
lipids were extracted using the method of Bligh and
Dyer (12), the extract was dissolved in cylcohexane,
and conjugated dienes were measured at 233 nm using
an extinction coefficient of 27,000 (mol/L)-1cm-1.
Lipid composition analyses. Liver lipids were extracted
with chloroform/methanol (2:1) according to the
method of Bligh and Dyer (12). Total cholesterol was
determined according to Zlatkis et al. (13), whereas
HDL cholesterol was determined using the method of
Lopes-Virella et al. (14). Triglycerides were assayed
according to Gottfried and Rosenberg (15).
Phospholipids were determined using the method of
Fiske and Subbarow (16) after digesting with
perchloric acid.
Plasma glucose determination. Glucose was determined
using the method of Teller (17).
Transaminase assay. The activities of alanine and
aspartate transaminases were determined using the
method of Reitman and Frankel (18) as reported by
Bergmeyer and Erlich (19).
Acid phosphatase assay. The activity of acid phosphatase
was determined using the method of Calzyme
Laboratories (20).

Statistical analysis
Data were analyzed statistically by Duncans Multiple
Range Test. Differences between groups were
considered significant at p<0.05 (21).

M. A. McAnuff, F. O. Omoruyi, E. Y. Morrison, H. N. Asemota / HEPATIC FUNCTION ENZYMES AND LIPID PEROXIDATION IN
STREPTOZOTOCIN-INDUCED DIABETIC RATS FED BITTER YAM (DIOSCOREA POLYGONOIDES)
STEROIDAL SAPOGENIN EXTRACT

Table 1. Feed intake, body and liver weight (g) changes in diabetic rats fed yam sapogenin extract or commercial
diosgenin
Normal
control

Diabetic control

Sapogenin extract

Commercial
diosgenin

Fi nal b od y w ei g ht

325.4 9.2a

247.3 27.0b

190.8 17.4b

193.3 8.8b

Ini ti al b od y w ei g ht

248.4 16.2a

249.3 9.2a

249.8 14.1a

248.4 16.8a

Dai ly feed i ntake (p er rat)

14.6 0.0

B lood g lucose after feed i ng (mMol)

5.87 0.38

14.5 0.1

29.60 2.20

7.5 0.4a

Li ver w ei g ht

14.6 0.1
b

18.20 0.71

6.6 0.6a

14.4 0.1a
b

19.72 0.77c

7.2 0.6a

6.3 0.4a

Values are means SEM; values i n the same row w i th d i fferent sup erscri p t are stati sti cally si g ni fi cant (p >0.05)

Table 1 shows food intake, body and liver weight


changes, and final blood glucose in diabetic rats fed
yam sapogenin extract or commercial diosgenin. The
diabetic rats (fed supplemented and unsupplemented
diets) lost weight significantly compared with the
normal group, although there was no significant
difference in their food intake. The decrease in body
weight was more significant in rats fed sapogenin
extract or commercial diosgenin. There was no
significant change in liver weight. Blood glucose was
significantly elevated in diabetic rats compared with
the normal controls. Supplementation of diets with
bitter yam sapogenin extract and commercial diosgenin
significantly decreased blood glucose towards normal.
The liver cholesterol level is shown in Table 2. The
feeding of supplemented diets to diabetic rats resulted
in a significant decrease in total cholesterol and VLDLcholesterol levels, and a significant increase in HDLcholesterol levels.
Table 2. Influence of yam steroidal sapogenin extract
and commercial diosgenin on liver cholesterol levels
Total
HDLcholesterol
cholesterol VLDL + LDL
( mol/g liver) ( mol/g liver) ( mol/g liver)
8.06 0.51

4.15 0.54

1.81 0.78

Diabetic

13.99 0.78

3.37 0.54

7.25 0.78

S apogenin
extract

11.40 0.78

6.70 0.54

1.8 0.58

Commercial
diosgenin

11.92 1.04

6.22 0.54

2.85 0.26

Normal

Values are means SEM; values i n the same row w i th d i fferent


sup erscri p t are stati sti cally si g ni fi cant (p >0.05)

Diabetologia Croatica 32-1, 2003

Figure 1. Influence of yam steroidal sapogenin extract


and commercial diosgenin on liver triglyceride and
phospholipid levels
0,004

phospholipids

triglycerides

0,0035

a
a

mol/g
mol/g liver
liver (wet
(wetwt.)
wt.)

RESULTS

0,003
a
a
a

0,0025

0,002

0,0015

0,001

0,0005

Normal

Diabetic

Sapogenin
extract

Commercial
diosgenin

Figure 1 shows the effect of yam steroidal sapogenin


extract and commercial diosgenin on the phospholipid
and triglyceride levels. Triglyceride and phospholipid
concentrations did not differ in normal and diabetic
rats. These variables were not significantly affected by
the dietary supplements.
Table 3 shows the effect of yam steroidal sapogenin
extract and commercial diosgenin on the HDLcholesterol and phospholipid to total cholesterol ratio.
The ratio of HDL-cholesterol and phospholipids to
total cholesterol was significantly lowered in diabetic
rats compared with the normal group. Supplementation of the diet with sapogenin extract and
commercial diosgenin resulted in a significant increase
in their ratios, restoring them almost to that of the
normal group.

19

M. A. McAnuff, F. O. Omoruyi, E. Y. Morrison, H. N. Asemota / HEPATIC FUNCTION ENZYMES AND LIPID PEROXIDATION IN
STREPTOZOTOCIN-INDUCED DIABETIC RATS FED BITTER YAM (DIOSCOREA POLYGONOIDES)
STEROIDAL SAPOGENIN EXTRACT

Table 3. Effect of yam steroidal sapogenin extract and


commercial diosgenin on the ratio of HDL-cholesterol
and phospholipids to total cholesterol
HDL cholesterol to Molar cholesterol:
total cholesterol
molar
ratio (%)
phospholipid ratio
Normal

45.7 2.1

0.13

Diabetic

24.1 1.8

0.23

S apogenin
extract
Commercial
diosgenin

61.4 2.8

52.2 2.3

0.14

0.16

Table 5. Effect of bitter yam steroidal sapogenin


extract, commercial diosgenin and bitter yam on liver
function enzymes
Alanine
transaminase

Normal

Table 4 shows the amount of lipid peroxidation


products in the liver. Bitter yam sapogenin extract and
diosgenin had significant effects on the hepatic levels
of oxidation products. Diabetic rats receiving the
unsupplemented diet had significantly (p<0.05)
higher levels of conjugated dienes (Table 4) and
TBARS than the supplemented groups. Diabetic rats
also had significantly higher levels of conjugated dienes
than normal rats.
Table 4. Lipid peroxidation products (TBARS and
conjugated dienes) in the livers of rats fed bitter yam
sapogenin extract or commercial diosgenin
Conjugated dienes
( mole/g liver)

TBARS
(nmol/g liver)

Normal

0.495 0.040

26.1 2.9

Diabetic

0.582 0.068

31.3 2.2

S apogenin
extract

0.416 0.023

17.7 1.1

Commercial
diosgenin

0.428 0.082

18.1 2.1

Values are means S EM; figures in the same row with different
superscript are statistically significant (p>0.05)

Table 5 shows the effect of bitter yam steroidal


sapogenin extract and commercial diosgenin on liver
function enzymes. Alanine and aspartate transaminase
activities were significantly increased in the livers of
diabetic rats (group B) compared with that of the
normal group. Supplementation of the diet with yam
steroidal sapogenin extract and commercial diosgenin
resulted in a significant decrease in the alanine

Acid
phosphatase

Aspartate
transaminase

S p.Act.=mg pyr/min/mg protein x 10

Values are means S EM; values in the same row with different
superscript are statistically significant (p>0.05)

20

transaminase activity. Neither the food supplements


nor the induction of diabetes had any effect on the
activity of acid phosphatase.

16.9 2.15 a
b

S p. Act. = mg
PNP/min/mg
protein x 10-5

-4

9.1 1.5

16.2 0.5 a

10.4 1.8

17.9 0.5 a

Diabetic

22.4 4.14

S apogenin
extract

12.6 1.80 a

8.5 0.7ab

17.9 0.6

Commercial
diosgenin

14.1 2.80 a

8.6 0.7ab

17.3 0.9 a

Values are means SEM; values i n the same row w i th d i fferent


sup erscri p t are stati sti cally si g ni fi cant (p >0.05)

DISCUSSION
The feeding of 1% diosgenin to hypercholesterolemic
rats has been shown to significantly lower plasma and
liver lipids (22). In this study, the short-term feeding of
bitter yam steroidal sapogenin extract resulted in
significantly decreased liver total cholesterol and
VLDL cholesterol levels, and significantly increased
HDL-cholesterol level (22,23). The increase in HDLcholesterol levels may be beneficial owing to the
negative correlation between HDL-cholesterol levels
and cardiovascular diseases. The steroidal sapogenin
extract was more effective in decreasing liver lipid
distribution than commercial diosgenin. This could be
due to the presence of other hypolipidemic agents such
as -sitosterol (24) present in the bitter yam steroidal
sapogenin extract.
There was no significant change in triglyceride and
phospholipid levels between diabetic and normal
groups (25). The cholesterol to phospholipid ratio was
significantly increased in the diabetic group compared
with normal rats. This may be due to the increased
cholesterol levels observed in the diabetic group and
the lack of change in the phospholipid levels. Increased
cholesterol to phospholipid ratio may affect the motion
of the lipid hydrocarbon chains and physical properties
of the bilayer (26). The observed hypocholesterolemic
properties of yam steroidal sapogenin extract and
commercial diosgenin significantly decreased the

M. A. McAnuff, F. O. Omoruyi, E. Y. Morrison, H. N. Asemota / HEPATIC FUNCTION ENZYMES AND LIPID PEROXIDATION IN
STREPTOZOTOCIN-INDUCED DIABETIC RATS FED BITTER YAM (DIOSCOREA POLYGONOIDES)
STEROIDAL SAPOGENIN EXTRACT

cholesterol to phospholipid ratio towards that of normal


rats. Ulloa and Nervi (27) have reported that steroid
molecules may modify the cholesterol-phospholipid
interactions in the membrane by interacting with the
hydrophobic part of the membrane. This modification
may be beneficial as increases in the levels of
cholesterol render membranes more lipophilic and
therefore less easy to degrade (28).
Conjugated dienes absorb ultraviolet light at 230235
nm and are considered by researchers appropriate for
the measurement of lipid peroxidation (29). The levels
of conjugated dienes and TBARS in the liver measure
its susceptibility to lipid peroxidation as well as the
antioxidant capability of dietary supplements. In this
study, the highest levels of TBARS and conjugated
dienes were seen in the diabetic group, suggesting
them to be more susceptible to lipid peroxidation
compared with normal rats. This result is consistent
with numerous reports of increased oxidative stress in
the tissues of diabetic rats (30-35). Supplementation of
the diet with bitter yam steroidal sapogenin extract or
commercial diosgenin reduced the diabetes-induced
increase in the liver lipid peroxide levels, and thus
reduced the susceptibility of the liver to lipid
peroxidation. The effect of bitter yam steroidal
sapogenin extract on the activities of the antioxidant
enzymes (e.g., superoxide dismutase, glutathione
peroxidase, and catalase) is not known. However, these
enzymes are involved in the scavenging/inactivation of
the reactive oxygen species or redox metal ions before
lipid peroxidation takes place, and their assessment in

relation to this dietary supplement may elucidate the


probable mechanism of action in reducing lipid
peroxidation as observed in this study (36).
Recent publications have shown that saponins
extracted from soybeans inhibited the formation of
lipid peroxides in corn oil samples, owing to their
ability to scavenge free radicals (36). The antioxidative
activity of the sapogenin extract and commercial
diosgenin may also be due to the presence of the
hydroxyl groups similar to that reported for sesamol and
sesamolinol (37). The sapogenin extract used in this
study consisted of -sitosterol, stigmasterol
pennogenin, and diosgenin compounds, which contain
more hydroxyl groups compared with the commercial
diosgenin, and this may have accounted for the higher
antioxidant activity in the sapogenin extract group.
Alanine and aspartate transaminase activities are used
as an indicator of hepatocyte damage (38). Acid
phosphatase activity is normally high in diseased states
and is often used as a tool in clinical investigations
(39). Data from this study show that alanine
transaminase activity is elevated in diabetes, while the
feeding of the sapogenin extract and commercial
diosgenin resulted in a significant decrease in alanine
transaminase activity. These dietary supplements may
protect liver cells from free radical damage.
In conclusion, the study has demonstrated that the
feeding of yam sapogenin extract and commercial
diosgenin to diabetic rats may lead to alterations in the
lipid composition of the liver and reduction in lipid
peroxidation, and may prevent liver damage associated
with diabetes.

REFERENCES
1.

Mandrup-Poulsen T. Recent advances in diabetes.


BMJ 1998;316:1221-1225.

2.

Javitt JC, Chiang Y. Economic impact of diabetes.


In: National Diabetes Data Group, eds. Diabetes
in America. Bethesda, MD: National Institutes of
Health, 1995; pp 233-255.

3.

Ragoobirsing D, Lewis-Fuller E, Morrison EY. The


Jamaican diabetes survey: a protocol for the
Caribbean. Diabetes Care 1995;18:1277-1279.

Diabetologia Croatica 32-1, 2003

4.

Giugliano D, Ceriello A, Paolisso G. Diabetes


mellitus, hypertension, and cardiovascular disease:
which role for oxidative stress? Metabolism
1995;44:363-368.

5.

Kakkar R, Kalra J, Mantha SV, Prasad K. Lipid


peroxidation and activity of antioxidant enzymes in
diabetic rats. Mol Cell Biochem 1995;151:113-119.

6.

Ozdemirler G, Mehmetcik G, Oztezcan S, Toker


G, Sivas A, Uysal M. Peroxidation potential and
antioxidant activity of serum in patients with

21

M. A. McAnuff, F. O. Omoruyi, E. Y. Morrison, H. N. Asemota / HEPATIC FUNCTION ENZYMES AND LIPID PEROXIDATION IN
STREPTOZOTOCIN-INDUCED DIABETIC RATS FED BITTER YAM (DIOSCOREA POLYGONOIDES)
STEROIDAL SAPOGENIN EXTRACT

diabetes mellitus and myocardial infarction. Horm


Metab Res 1995;27:194-196.
7.

8.

9.

Kaimal A, Kemper K. Wild yam (Dioscoreaceae)


Longwood Herbal Task Force 1999: http://
www.mcp.edu/herbal/default.htm
Coursey DG. Yams, an account of the nature,
origins, cultivation and utilization of the useful
members of Dioscoreaceae. London: Longman,
1967; pp 230.

19. Bergmeyer HU, Erlich B, eds. Methods of


enzymatic analysis. Weinheim New York: Verlag
Chemie Weinheim Academic Press, Inc., 1974;
pp 737-764.
20. Calzyme Laboratories http://www.calzyme.com/
catalog/acidphos.html
21. Sokal RR, Rohlf FJ. The principles and practice of
statistics in biological research. San Francisco, CA:
Freeman and Co., 1969; pp 469-484.

Morris MP, Roark BA, Cancel B. Simple procedure


for the routine assay of Dioscorea tubers. J Agric
Food Chem 1958;6:856-858.

22. Cayen M, Dvornik D. Combined effects of


clofibrate and diosgenin on cholesterol metabolism
in rats. Atherosclerosis 1978;29:317-328.

10. Tappel AL, Zalkin H. Inhibition of lipid


peroxidation in mitochondria by vitamin E. Arch
Biochim Biophys 1959;80:333-336.

23. Uchida K, Yakase H, Takeda K, Takeuchi N,


Ishikawa Y. Changes in biliary fecal bile acids in
mice after treatments with diosgenin and sitosterol. J Lipid Res 1984;25:236-245.

11. Hu M, Frankel EN, Leibovitz BE, Tappel AL.


Effect of dietary lipids and vitamin E on the in vitro
lipid peroxidation in rat liver and kidney
homogenates. J Nutr 1989;119:1574-1582.
12. Bligh EG, Dyer WJ. A rapid method of total lipid
extraction and purification. Can J Biochem Physiol
1959;37:911-917.
13. Zlatkis A, Zak B, Boyle AJ. A new method for the
direct determination of serum cholesterol. J Lab
Clin Med 1953;41:486-492.
14. Lopes-Virella MF, Store P, Ellis S, Colwell AJ.
Cholesterol determination in the high-density
lipoprotein separated by three different methods.
Clin Chem 1977;23:882-885.
15. Gottfried SP, Rosenberg B. Improved manual
spectrophotometric procedure for determination
of serum triglycerides. Clin Chem 1973;19:10771078.
16. Fiske CH, Subbarow Y. The colorimetric
determination of phosphorus. J Biol Chem
1925;66:375-400.
17. Teller JD. Direct colorimetric determination of
serum and plasma glucose. Abstracts of papers,
130th Meeting. American Chemical Society,
Washington DC, 1956;69C.
18. Reitman S, Frankel S. Colorimetric method for
determination of serum GOT and GPT. Am J Clin
Pathol 1957;28:56.

22

24. Ram A, Lauria P, Gupta R, Kumar P, Sharma VN.


Hypocholesterolaemic effects of Terminalia Arjuna
tree bark. J Ethnopharmacol 1997;55:165-169.
25. Kabir M, Rizkalla SW, Champ M et al. Dietary
amylase-amylopectin starch content affects
glucose and lipid metabolism in adipocytes of
normal and diabetic rats. J Nutr 1998;128:35-43.
26. Johnson SM. A new specific cholesterol assay gives
reduced cholesterol/phospholipid molar ratios in
cell membranes. Ann Biochem 1979;95:344-350.
27. Ulloa N, Nervi F. Mechanism and kinetic
characteristics of the uncoupling of plant steroids
of biliary cholesterol from bile salt output. Biochim
Biophys Acta 1985;837:181-189.
28. Patton S. Correlative relationship of cholesterol
and sphingomyelin in cell membranes. J Theor
Biol 1970;29:489-491.
29. Corongiu FP, Poli G, Dianzani MU. Lipid
peroxidation and molecular damage to
polyunsaturated fatty acids in rat liver: recognition
of two classes of hydroperoxides formed under
conditions in vivo. Chem Biol Interact
1986;59:147-155.
30. Wolfe S, Dean L. Glucose autoxidation and protein
modification. The potential role of autoxidative
glycosylation in diabetes. Biochem J 1987;245:
243-250.

M. A. McAnuff, F. O. Omoruyi, E. Y. Morrison, H. N. Asemota / HEPATIC FUNCTION ENZYMES AND LIPID PEROXIDATION IN
STREPTOZOTOCIN-INDUCED DIABETIC RATS FED BITTER YAM (DIOSCOREA POLYGONOIDES)
STEROIDAL SAPOGENIN EXTRACT

31. Baynes JW. Role of oxidative stress in the


development of complications in diabetes.
Diabetes 1991;40:401-402.
32. Bell RC, Carlson JC, Storr KC, Herbert K, Sivak J.
High-fructose feeding of streptozotocin-diabetic
rats is associated with increased cataract formation
and increased oxidative stress in the kidney. Br J
Nutr 2000:84:575-582.
33. Wohaieb SA, Godin DV. Alterations in free radical
tissue-defense mechanisms in streptozotocininduced diabetes in rats. Diabetes 1987;36:10141018.
34. Rerup CC. Drugs producing diabetes through
damage of the insulin secreting cells. Pharmacol
Rev 1970;22:485-518.
35. Lubec B, Hermon M, Hoeger H, Lubec G.
Aromatic hydroxylation in animal models of
diabetes mellitus. FASEB J 1998;12:1581-1587.

Diabetologia Croatica 32-1, 2003

36. Sung MK, Park MY. Effect of soybean saponins on


the growth and antioxidant defense of human
hepatocarcinoma cells.
Third International
Symposium on the Role of Soy in Preventing and
Treating Chronic Disease. October 31 - November
3, 1999. Omni Shoreham Hotel Washington, DC
USA D-3.
37. Kang M, Naito M, Tsujihara N, Osawa T.
Sesamolin inhibits lipid peroxidation in rat liver
and kidney. J Nutr 1998;128:1018-1022.
38. Whitehead MW, Hawkes ND, Hainsworth I,
Kingham JGC. A prospective study of the causes of
notably raised aspartate aminotransferase of liver
origin. Gut 1999;45:129-133.
39. Bull Murray PG, Thomas D, Fraser AM, Nelson
PN. Acid phosphatases. Mol Pathol 2000;55:65-72.
Acknowledgment. The authors wish to thank Graduate
Studies and Research, University of the West Indies,
Mona Campus, for providing financial support.

23