Bioscience, Biotechnology, and Biochemistry

ISSN: 0916-8451 (Print) 1347-6947 (Online) Journal homepage: https://www.tandfonline.com/loi/tbbb20

The Effect of Pre-germinated Brown Rice Intake on

Blood Glucose and PAI-1 Levels in Streptozotocin-
induced Diabetic Rats

Hiromi HAGIWARA, Taiichiro SEKI & Toyohiko ARIGA

To cite this article: Hiromi HAGIWARA, Taiichiro SEKI & Toyohiko ARIGA (2004) The Effect
of Pre-germinated Brown Rice Intake on Blood Glucose and PAI-1 Levels in Streptozotocin-
induced Diabetic Rats, Bioscience, Biotechnology, and Biochemistry, 68:2, 444-447, DOI: 10.1271/
bbb.68.444

To link to this article: https://doi.org/10.1271/bbb.68.444

Published online: 22 May 2014.

Submit your article to this journal

Article views: 620

View related articles

Citing articles: 117 View citing articles

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=tbbb20
Biosci. Biotechnol. Biochem., 68 (2), 444–447, 2004
Note
The Effect of Pre-germinated Brown Rice Intake on Blood Glucose
and PAI-1 Levels in Streptozotocin-induced Diabetic Rats
Hiromi H AGIWARA, Taiichiro S EKI,y and Toyohiko ARIGA
Laboratory of Nutrition and Physiology, Department of Agricultural and Biological Chemistry,
Nihon University College of Bioresource Sciences, Nihon University Graduate School of Bioresource Sciences,
Kanagawa 252-8510, Japan
Received August 8, 2003; Accepted October 29, 2003
Effects of pre-germinated brown rice (PGBR) on
streptozotocin-induced diabetic rats were studied. The
feeding of a PGBR diet to diabetic rats ameliorated the
elevation of blood glucose and PAI-1 concentrations
significantly, and tended to decrease the plasma lipid
peroxide concentrations in comparison with rats fed a
white rice diet. These results suggest that intake of
PGBR instead of white rice is effective for the pre-
vention of diabetic vascular complications.
Key words: pre-germinated brown rice; white rice; type-
1 plasminogen activator inhibitor (PAI-1);
diabetes
Diabetic macrovascular complications associated with
hyperglycemia and hyperlipidemia have been demon-
strated to be linked with the mortality among diabetic
patients.1) An increased plasma concentration of type-1
plasminogen activator inhibitor (PAI-1) has been ob-
served in type 2 diabetic patients with vascular dis-
eases.2) PAI-1 is the major physiological inhibitor of
tissue-type PA (tPA) and urokinase-type PA (uPA).
Both tPA and uPA convert a zymogen plasminogen into
an active enzyme, plasmin. Impaired fibrinolytic poten-
tial due to the increased level of plasma PAI-1 has been
shown to be a major risk factor for myocardial infarction
in diabetes mellitus.3) Increased oxidative stress in the
hyperglycemia has also been recognized as a causative
of vascular damage.4) Thus the hyperglycemia is thought
to be one of the important risk factors for vascular
complications in diabetes mellitus.
Pre-germinated brown rice (PGBR) has recently been
widely served in Japan. PGBR had been developed
industrially in order to improve the nutritional functions
of its source material, brown rice. PGBR can be
produced by soaking brown rice in water for slight
germination. Amounts of some constituents including
-
aminobutyric acid (GABA) are greatly increased in
PGBR. It has been reported that GABA and its related
y
To whom correspondence should be addressed. Fax: +81-466-84-3949; E-mail: tseki@brs.nihon-u.ac.jp
Abbreviations: PGBR, pre-germinated brown rice; PAI-1, type-1 plasminogen activator inhibitor; tPA, tissue-type plasminogen activator; uPA,
urokinase-type plasminogen activator; TBA, thiobarbituric acid; LDL, low density lipoprotein
enzymes are located in pancreatic beta cells5) and that
GABA stimulates insulin secretion from the pancreas.5)
Thus PGBR or its ingredient is thought to ameliorate the
disorders such as diabetes, although there is little
scientific evidence regarding nutritional function.
In this study the effects of long-term intake of PGBR
on the diabetic parameters was examined in comparison
with the intake of white rice. We have especially
focused on the effects of PGBR on plasma PAI-1
concentration which contributes to increase the mortal-
ity rate of diabetic patients, and found the facts that as
compared to white rice, PGBR intake could ameliorate
hyperglycemia and concomitant increase of plasma PAI-
1 concentrations in the experimentally-induced diabetic
rats.
All animal experiments were done in accordance with
institutional guidelines established by the Laboratory
Animal Care and Use Committee of Nihon University
College of Bioresource Sciences. Male Wistar strain rats
(Nippon Bio-Suppl., Tokyo, Japan), weighing 100 g,
received intraperitoneal injection of streptozotocin
(65 mg/kg, dissolved in 100 mmol/l sodium citrate
buffer, pH 4.5; Wako Pure Chemical Inc. Ltd., Osaka,
Japan). Control rats received only the same volume of
sodium citrate buffer. The rats were kept on a standard
diet (CE-2, CLEA JAPAN Inc., Tokyo, Japan) for 13
days after the streptozotocin injection, and then divided
into four groups: Non-diabetic rats fed white rice diet
(NW), non-diabetic rats fed PGBR diet (NB), diabetic
rats fed white rice diet (DW), diabetic rats fed PGBR
diet (DB). These rats were allowed free access to the
experimental diets and water for 7 weeks. AIN-93G
(Oriental Yeast Co., Ltd., Tokyo, Japan) containing 53%
corn-starch was used as basic diet. Corn-starch of AIN-
93G was replaced with white rice powder for the ‘‘white
rice diet’’ or replaced with PGBR powder (FANCL Co.,
Kanagawa, Japan) for the ‘‘PGBR diet’’. White rice
powder contained (per 100 g of powder) water, 15.8 g;
protein, 5.7 g; fat, 1.7 g; carbohydrate, 75.7 g; dietary
 Regulation of Plasma PAI-1 Level by Pre-germinated Brown Rice
fiber, 0.6 g; Ca, 4.7 mg; Mg, 32.2 mg; thiamine, 0.11 mg;
total tocopherols, 0.3 mg; and free
-aminobutyric acid,
3 mg. PGBR powder contained (per 100 g of powder)
water, 14.7 g; protein, 6.9 g; fat, 2.5 g; carbohydrate,
72 g; dietary fiber, 2.7 g; Ca, 8.8 mg; Mg, 117 mg;
thiamine, 0.37 mg; total tocopherols, 1.3 mg; free
-
aminobutyric acid, 15 mg. Amounts of food intake and
body weight were measured every day, and blood
glucose was measured by DEXTER-Z II (Bayer Medical
Co., Ltd., Leverkusen, Germany) once a week using
blood obtained from the tail vein. The rats were starved
for 18 hours before the measurement of blood glucose
concentration. Serum triglyceride concentration was
measured by using a commercial kit (Wako Pure
Chemical Inc., Ltd.). Plasma PAI-1 concentration was
measured by an ELISA system using anti-rat PAI-1
rabbit IgG (American Diagnostica Inc., CT, USA) and
recombinant rat PAI-1 (American Diagnostica Inc.) as
standards. The plasma lipid peroxide concentration was
measured by SRL Inc. (Tokyo, Japan) using thiobarbi-
turic acid (TBA) method. Serum insulin concentration
was measured by a commercial ELISA kit (LBISTM ,
Shibayagi Co., Ltd., Gunma, Japan).
Total amounts of food intake during the 7-weeks
experimental period were not significantly different
between the white rice group and the PGBR group in
both normal and diabetic rats, however, the rats tended
to prefer the PGBR diet rather than white rice diet
(Table 1). The modest body weight gain which would be
due to the metabolic disorder with diabetes was
observed in the diabetic group, but there was no
significant difference between white rice and PGBR
diet groups in both normal and diabetic groups (Table 1).
The serum triglyceride level of the diabetic group was
increased as compared to that of normal group, although
there was no significant difference among the groups
(Table 1).
Blood glucose concentrations were mostly equal
between normal rat groups, NW and NB, throughout
the experimental period. Whereas in the diabetic group,
blood glucose concentration was lower in the rat fed
PGBR (DB) than that in white rice fed rats (DW)
(Fig. 1). These results are very similar to those obtained
Table 1. Total Amount of Food Intake, Body Weights and Serum
Triglyceride of Rats Fed Either White Rice or Pre-germinated Brown
Rice
Total amount of Body weight (g) Triglyceride (mg/dl)
food intake (g)
0 time 7 weeks 7 weeks
NW 1387
34 178.0
3.2 532.4
14.5 282.1
54.7
NB 1422
46 179.4
2.5 530.3
15.3 308.4
31.9
DW 1496
63 123.0
7.0 156.5
16.7 446.0
92.0
DB 1546
30 119.5
6.1 167.7
13.5 320.8
39.8
NW, normal rats fed white rice diet (n=8); NB, normal rats fed PGBR diet
(n=7); DW, diabetic rats fed white rice diet (n=4); DB, diabetic rats fed
PGBR diet (n=5). Triglyceride concentration was measured by a commer-
cial kit as described in the text. Each value represents the mean
S.E.
445
500
400
*
NW
300
NB
mg/dl
DW
200 DB
100
0
0 1 2 3 4 5 6 7 Weeks
Fig. 1. Blood Glucose Concentration of Rats Fed Either White Rice
or PGBR.
NW, normal rats fed white rice diet (n=8); NB, normal rats fed
PGBR diet (n=7); DW, diabetic rats fed white rice diet (n=4); DB,
diabetic rats fed PGBR diet (n=5). Blood glucose concentrations
were measured as described in the text. Each value represents the
mean
S.E. Absence of error bars for a given data point indicated
that S.E. is smaller than the size of symbol. The statistical analysis
was done by the Student’s t-test. *P < 0:05 vs. DW.
from a human study; the glycemic index obtained from
volunteers taking PGBR was significantly lower than
that of those who were taking the white rice (unpub-
lished data). The blood glucose-lowering effect of
PGBR may be derived from the properties of PGBR
involving substantially higher content of dietary fiber
than white rice (about 5-fold, as described before), and
PGBR exhibits the lower postprandial glycemic re-
sponse. Intake of barley containing high dietary fiber is
known to reduce the fasting plasma glucose, plasma
cholesterol, and triglyceride levels in spontaneous
diabetic rat.6) Therefore, the tendency to lower the
serum triglyceride elevation in DB group may also be
derived from dietary fiber in PGBR.
Plasma PAI-1 concentrations in normal rats fed any
diet (NW or NB) were approximately 0.9 ng/ml
(Fig. 2A). As reported previously in both type 17) and
type 2 diabetic rats,8) plasma PAI-1 concentration was
elevated in the diabetic rats. The feeding of white rice to
diabetic rats caused significantly higher plasma PAI-1
concentrations than normal rats fed white rice (Fig. 2A,
NW vs. DW), however the feeding of PGBR signifi-
cantly decreased the PAI-1 concentration of the diabetic
rats (DW vs. DB). Changes in the plasma lipid peroxide
concentration showed a similar tendency to those
observed in plasma PAI-1 concentration (Fig. 2B).
Increased PAI-1 concentration results in impairment of
fibrinolysis as has been reported in diabetic patients.9)
Since plasma PAI-1 level can be modulated by blood
glucose concentration and hypertriglyceridemia,10,11)
intake of PGBR may reduce the risk of ischemic heart
disease including myocardial infarction in diabetes
mellitus. We have recently reported that treatment of
the type-1 diabetic rats with insulin reduced the up
regulated-plasma PAI-1 concentration.7) Plasma insulin
446 H. HAGIWARA et al.

A P<0.01 P<0.01 B P<0.01

8 30
7 25
6
20

nmol/ml
5
ng/ml

4 15
3 10
2
1 5
0 0
NW NB DW DB NW NB DW DB

Fig. 2. Plasma PAI-1 and Lipid Peroxide Concentration of Rats Fed Either White Rice or PGBR.
(A) Plasma PAI-1 concentration measured by ELISA. (B) Plasma lipid peroxide concentration measured by TBA method. NW, normal rats
fed white rice diet (n=8); NB, normal rats fed PGBR diet (n=7); DW, diabetic rats fed white rice diet (n=4); DB, diabetic rats fed PGBR diet
(n=5). Each value represents the mean
S.E. Two-way factorial ANOVA was used to evaluate the differences between normal and diabetic
groups, and white rice and PGBR groups. Because a significant interaction was observed between the groups, one-way ANOVA and following
Turkey HSD test as post-ANOVA comparisons were done.

concentration was also measured in this study; NW,
1.51
0.53; NB, 1.04
0.35; DW, 0.29
0.07; DB,
0.62
0.15 ng/ml. In the diabetic rats, the plasma insulin
concentration was correlated with plasma PAI-1 and
glucose concentrations.
Oxidative stress occurring as a consequence of
hyperglycemia is known to play a central role in the
pathogenesis and progression of diabetes and its com-
plication.12) In this study, the plasma lipid peroxide
concentration was significantly increased in the diabetic
rats fed white rice (DW) in comparison with normal rats
fed either white rice or PGBR. In the diabetic rats,
feeding of PGBR tended to decrease the plasma lipid
peroxide concentration in comparison with the feeding
of white rice, although there was no statistic significance
between both groups (DW vs. DB). This peroxide-
lowering effect is considered to be derived from not only
antioxidative vitamin (tocopherol) which is contained
highly in PGBR but also other factors effective in
preventing the increase in blood glucose concentration.
Moreover, it has been reported that the oxidized LDL
induces PAI-1 production in vascular endothelial
cells.13) Therefore, the antioxidant effect of PGBR
would be one of the factors in preventing the elevation
of plasma PAI-1 concentration in DB group. Taken
together, the findings obtained from this study indicate
that there is potential benefit from the intake of PGBR
instead of white rice in the prevention of diabetic
vascular complications such as ischemic heart disease.
In summary, PGBR ameliorated the blood glucose,
plasma PAI-1, and lipid peroxide concentrations in
diabetic rats in comparison with these parameters of
diabetic rats fed white rice. These effects might be
derived from the properties of PGBR that include
substantially higher contents of functional components
such as tocopherols, dietary fiber, GABA and other
unknown factor(s), and from the property to exhibit the
lower postprandial glycemic response.
The authors thank Ms Rieko Nakane and Mr Chikao
Uchiyama for the animal experiments, and Mr Mitsuo
Kise, Ms Aya Mizukuchi, and Mr Yukihiko Ito (FANCL
Co.) for providing pre-germinated brown rice.
References
1) Sobel, B. E., Effects of glycemic control and other
determinants on vascular disease in type 2 diabetes. Am.
J. Med., 113, 12S–22S (2002).
2) Skrha, J., Hodinar, A., Kvasnicka, J., and Hilgertova, J.,
Relationship of oxidative stress and fibrinolysis in
diabetes mellitus. Diabet. Med., 13, 800–805 (1996).
3) Hamsten, A., de Faire, U., Walldius, G., Dahlen, G.,
Szamosi, A., Landou, C., Blomback, M., and Wiman, B.,
Plasminogen activator inhibitor in plasma: risk factor for
recurrent myocardial infarction. Lancet, 2, 3–9 (1987).
4) Gallou, G., Ruelland, A., Legras, B., Maugendre, D.,
Allannic, H., and Cloarec, L., Plasma malondialdehyde
in type 1 and type 2 diabetic patients. Clin. Chim. Acta,
214, 227–234 (1993).
5) Adeghate, E., and Ponery, A. S., GABA in the endocrine
pancreas: cellular localization and function in normal
and diabetic rats. Tissue Cell, 34, 1–6 (2002).
6) Li, J., Kaneko, T., Wang, Y., Qin, L. Q., and Sato, A.,
Effects of dietary fiber on the glucose tolerance in
spontaneously diabetic rats—comparison among barley,
rice, and corn starch. Nippon Eiseigaku Zasshi (in
Japanese), 58, 281–286 (2003).
7) Hagiwara, H., Kaizu, K., Uriu, K., Noguchi, T., Takagi,
I., Qie, Y. L., Seki, T., and Ariga, T., Expression of type-
1 plasminogen activator inhibitor in the kidney of
diabetic rat models. Thromb. Res., 111, 301–309 (2003).
8) Uehara, Y., Hirawa, N., Numabe, A., Kawabata, Y.,
Nagoshi, H., Negoro, H., Fujiwara, S., Gomi, T., Ikeda,
T., Goto, A., and Omata, M., Angiotensin-converting
enzyme inhibition delays onset of glucosuria with
regression of renal injuries in genetic rat model of
non-insulin-dependent diabetes mellitus. J. Cardiovasc.
Pharmacol. Ther., 3, 327–336 (1998).
9) Kruszynska, Y. T., Yu, J. G., Olefsky, J. M., and Sobel,
B. E., Effects of troglitazone on blood concentrations of
plasminogen activator inhibitor 1 in patients with type 2
 Regulation of Plasma PAI-1 Level by Pre-germinated Brown Rice
diabetes and in lean and obese normal subjects.
Diabetes, 49, 633–639 (2000).
10) Vukovich, T., Proidl, S., Knobl, P., Teufelsbauer, H.,
Schnack, C., and Schernthaner, G., The effects of insulin
treatment on the balance between tissue plasminogen
activator and plasminogen activator inhibitor-1 in type 2
diabetic patients. Thromb. Haemost., 68, 253–256
(1992).
11) Calles-Escandon, J., Mirza, S. A., Sobel, B. E., and
Schneider, D. J., Induction of hyperinsulinemia com-
447
bined with hyperglycemia and hypertriglyceridemia
increases plasminogen activator inhibitor 1 in blood in
normal human subjects. Diabetes, 47, 290–293 (1998).
12) Baynes, J. W., Role of oxidative stress in development of
complications in diabetes. Diabetes, 40, 405–412 (1991).
13) Ren, S., Man, R. Y., Angel, A., and Shen, G. X.,
Oxidative modification enhances lipoprotein (a)-induced
overproduction of plasminogen activator inhibitor-1 in
cultured vascular endothelial cells. Atherosclerosis, 128,
1–10 (1997).