JOURNAL OF MEDICINAL FOOD

J Med Food 12 (4) 2009, 714–721

Full Communication
# Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition
DOI: 10.1089=jmf.2008.1282

Matcha, a Powdered Green Tea, Ameliorates the Progression of Renal

and Hepatic Damage in Type 2 Diabetic OLETF Rats
Noriko Yamabe, Ki Sung Kang, Jong Moon Hur, and Takako Yokozawa
Institute of Natural Medicine, University of Toyama, Toyama, Japan

ABSTRACT Matcha, a powdered green tea produced by grinding with a stone mill, has been popularly used in the
traditional tea ceremony and foods in Japan. Matcha is well known to be richer in some nutritional elements and epigallo-
catechin 3-O-gallate than other green teas. In our previous study, epigallocatechin 3-O-gallate exhibited protective effects
against renal damage in a rat model of diabetic nephropathy. In the present study, we investigated the preventive effects of
Matcha (50, 100, or 200 mg=kg=day) on the progression of hepatic and renal damage in type 2 diabetic Otsuka Long-Evans
Tokushima Fatty (OLETF) rats. OLETF rats were orally administered Matcha for 16 weeks, and we assessed biochemical
parameters in the serum, liver, and kidney and expression levels of major products of advanced glycation end products
(AGEs), Ne-(carboxylmethyl)lysine (CML) and Ne-(carboxylethyl)lysine (CEL), receptor for AGE (RAGE), and sterol reg-
ulatory element binding proteins (SREBPs)-1 and -2. Serum total protein levels were significantly increased by Matcha
administration, whereas the serum albumin and glycosylated protein levels as well as the renal glucose and triglyceride levels
were only slightly or not at all affected. However, Matcha treatment significantly lowered the glucose, triglyceride, and total
cholesterol levels in the serum and liver, renal AGE levels, and the serum thiobarbituric acid-reactive substances levels. In
addition, Matcha supplementation resulted in decreases in the renal CML, CEL, and RAGE expressions as well as an increase
in hepatic SREBP-2 expression, but not that of SREBP-1. These results suggest that Matcha protects against hepatic and renal
damage through the suppression of renal AGE accumulation, by decreases in hepatic glucose, triglyceride, and total cho-
lesterol levels, and by its antioxidant activities.

KEY WORDS:
advanced glycation end products
Matcha
sterol regulatory element binding proteins
type 2 diabetes

INTRODUCTION glycemia, dyslipidemia, AGEs and their receptors, and ox-

idative stress are strongly associated with the pathogenesis
T he incidence of type 2 diabetes has markedly in-
creased worldwide. Obesity is considered to be the
major risk factor in the development of type 2 diabetes be-
of diabetes and its complications, such as nephropathy and
cardiovascular diseases.5–9
The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is
cause 90% of type 2 diabetes patients are obese. Both type 2
an animal model of spontaneous obesity and type 2 diabetes
diabetes and obesity are closely associated with genetic
characterized by hyperglycemia, insulin resistance, hy-
factors, lifestyle, lack of physical exercise, and an increase in
perinsulinemia, hypertriglyceridemia, and hypercholester-
food intake.1,2 Reciprocally, an increase in the duration of
olemia, with complications such as nonalcoholic fatty liver
obesity is accompanied by an increase in the incidence of
and renal disorders.10,11 Hyperglycemia, hyperinsulinemia,
type 2 diabetes and deterioration of the glucose response.3,4
hypertriglyceridemia, and insulin resistance become marked
Hyperglycemia increases the formation of advanced glyca-
after the age of 18 weeks. In the hyperinsulinemic stage,
tion end products (AGEs) and reactive oxygen species, such
there are hyperplastic changes in the pancreatic islets. At 40
as superoxide, and decreases the NADPH and glutathione
weeks of age, the pancreatic islet mass is decreased and
levels in cells, which result in the increase of lipid peroxide
replaced by connective tissue. The rats finally become hypo-
levels, an indicator of oxidative stress.5–7 Therefore, hyper-
insulinemic and lose weight. These typical characteristics of
OLETF rats are known to be useful for investigating the
Manuscript received 12 September 2008. Revision accepted 28 April 2009. complex forms of human diabetes.10
Lifestyle modifications are considered important for the
Address correspondence to: Takako Yokozawa, Institute of Natural Medicine, University
of Toyama, 2630 Sugitani, Toyama 930-0194, Japan, E-mail: yokozawa@inm.u-toyama
prevention and treatment of obesity-related type 2 diabe-
.ac.jp tes and its complications, including antioxidant therapy,

714
 MATCHA IN TYPE 2 DIABETIC OLETF RATS 715

modulators of proteins, and genes associated with glucose shima, Japan). All rats were kept in wire-bottomed cages
and lipids along with inhibitors of AGEs and receptor for with controlled temperature (about 258C) and humidity
AGE (RAGE).8,12–14 We have reported the beneficial effects (about 60%) and a 12-hour light=dark cycle. Rats were given
of epigallocatechin 3-O-gallate, a major component of free access to laboratory pellet chow (CE-2, CLEA Japan,
Matcha, against oxidative renal damage in type 1 diabetic Inc., Tokyo) composed of 24.0% protein, 3.5% lipids, 60.5%
nephropathy model rats.15 Matcha has been considered a carbohydrate, and water. At 22 weeks of age rats were di-
precious and good source of green tea polyphenols and fiber vided into five groups: a nondiabetic LETO group given
in Japan, but its effects on both hepatic and renal dysfunc- water (n ¼ 5), a diabetic OLETF control group given water
tion are not yet fully elucidated. In the present study, we (n ¼ 10), and three groups (each n ¼ 10) of diabetic OLETF
investigated the effects of Matcha on hepatic and renal rats given Matcha (50, 100, or 200 mg=kg of body weight=
damage using a model of human type 2 diabetes mellitus, day) by oral gavage once a day. During the administration
the OLETF rat, to identify its roles in hyperglycemia, dys- period of 16 weeks, rats were housed in individual cages, and
lipidemia, AGEs and their receptors, and also oxidative food and water consumption was recorded. Blood samples
stress. were collected from tail veins for the determination of serum
glucose and total cholesterol levels every 4 weeks, and urine
MATERIALS AND METHODS samples were collected using metabolic cages every 8 weeks.
At the end of the experiment, all rats were sacrificed after an
Materials intraperitoneal injection of pentobarbital. Blood samples
Matcha, a powdered green tea, was kindly supplied by were collected from the abdominal aorta, and the liver and
Aiya Co., Ltd. (Aichi, Japan). It was composed mainly of kidneys were removed from each rat, rinsed with ice-cold
caffeine (3.0%), polyphenol (10.0%), protein (31.0%), physiological saline, weighed, and frozen at 808C in the
fiber (39.0%), calcium (0.42%), iron (0.02%), potassium refrigerator (Thermo Fisher Scientific K.K., Kanagawa,
(2.7%), vitamin A (0.005%), vitamin C (0.06%), and caro- Japan) until analysis.
tene (0.029%). 4,6-Dihydroxy-2-mercaptopyrimidine (2-
thiobarbituric acid [TBA]), 5-hydroxymethylfurfural, oxalic Clinical characteristics of animals
acid, bovine serum albumin (BSA), 2-amino-2-hydroxy- Serum levels of glucose, total protein, albumin, total
methyl-1,3-propadiol [Tris(hydroxymethyl)aminomethane], cholesterol, triglycerides, and creatinine (Cr) were exam-
Tween 20, phenylmethylsulfonyl fluoride (PMSF), protease ined using commercial reagents (Glucose CII-Test Wako,
inhibitor mixture dimethyl sulfoxide (DMSO) solution, and A=G B-Test Wako, Cholesterol E-Test Wako, and Trigly-
skim milk powder were purchased from Wako Pure Che- ceride E-Test Wako from Wako Pure Chemical Industries,
mical Industries, Ltd. (Osaka, Japan). Dithiothreitol was Ltd.; CRE-EN Kainos was obtained from Kainos Labora-
purchased from BioVision, Inc. (Mountain View, CA). The tories, Inc. [Tokyo]). Serum glycosylated protein was
Bio-Rad protein assay kit was purchased from Bio-Rad colorimetrically measured by determining 5-hydroxy-
Laboratories (Tokyo, Japan). Polyclonal anti-RAGE anti- methylfurfural formation from glucose according to the TBA
body (sc-5563), rabbit polyclonal anti-sterol regulatory assay.16 TBA-reactive substances (TBARS) levels were de-
element binding protein (SREBP)-1 antibody (sc-8984), termined using the methods of Naito and Yamanaka.17 Urine
rabbit polyclonal anti-SREBP-2 antibody (sc-5603), goat component levels were determined as follows: protein by the
anti-rabbit immunoglobulin G horseradish peroxidase- sulfosalicylic acid method and Cr using a commercial re-
conjugated secondary antibody (sc-2004), and goat agent (CRE-EN Kainos). Cr clearance (Ccr) was calculated
anti-mouse immunoglobulin G horseradish peroxidase- on the basis of urinary Cr, serum Cr, urine volume, and body
conjugated polyclonal anti-receptor secondary antibody (sc- weight using the following equation: Ccr (in mL=kg of body
2005) were purchased from Santa Cruz Biotechnology, Inc. weight=minute) ¼ (urinary Cr [in mg=dL]urinary volume
(Santa Cruz, CA). Monoclonal anti-Ne-(carboxyethyl)lysine [in mL]=serum Cr [in mg=dL])(1,000=body weight [in
(CEL) antibody and polyclonal anti-Ne-(carboxymethyl)- g])(1=1,440 [in minutes]).
lysine (CML) antibody were kindly provided by Dr. R. Nagai
(Kumamoto University, Kumamoto, Japan). Glycerol, Renal and hepatic glucose, AGE, total cholesterol,
Nonidet P-40, and anti-mouse b-actin antibody were pur- and triglyceride levels
chased from Sigma-Aldrich (St. Louis, MO). Enhanced
chemiluminescence western blotting detection reagents Renal and hepatic glucose levels were determined by the
were purchased from GE Healthcare (Piscataway, NJ). method of Momose et al.18 with minor modifications. In
brief, frozen kidney and liver tissues were homogenized
with ice-cold physiological saline and deproteinized, and
Animals
glucose concentration was determined using the Wako kit
The Guidelines for Animal Experimentation approved by described above. AGE levels in the kidney and liver were
the University of Toyama were followed in all experimental determined by the method of Nakayama et al.19 In brief,
studies. Male OLETF and Long-Evans Tokushima Otsuka each tissue was minced and then delipidated with chloro-
(LETO) rats were kindly supplied by the Tokushima Re- form and methanol (2:1, vol=vol) overnight. After washing,
search Institute (Otsuka Pharmaceutical Co., Ltd., Toku- the tissue was homogenized in 0.1 N NaOH, followed by
716 YAMABE ET AL.

centrifugation at 8,000 g for 15 minutes at 48C. The amounts protease inhibitor mixture DMSO solution. Then, 65 mL of
of AGEs in these alkali-soluble samples were determined by 2% (vol=vol) Nonidet P-40 was added, and the mixture was
measuring the fluorescence at an emission wavelength of vortex-mixed for 1 minute, incubated on ice for 10 minutes,
440 nm and excitation wavelength of 370 nm. A native BSA and centrifuged at 1,000 g for 10 minutes at 48C. Crude
preparation (1 mg=mL in 0.1 N NaOH) was used as a stan- nuclear pellets were rinsed twice with hypotonic buffer and
dard, and its fluorescence intensity was defined as 1 unit of resuspended in 60 mL of hypertonic buffer containing
fluorescence. The fluorescence values of samples were 50 mM HEPES (pH 7.9), 50 mM KCl, 300 mM NaCl, 10%
measured at a protein concentration of 1 mg=mL and ex- (vol=vol) glycerol, 0.5 mM dithiothreitol, 1 mM PMSF, and
pressed in arbitrary units (AU) compared with a native BSA protease inhibitor mixture DMSO solution. It was then
preparation. The kidneys and liver of each rat were ho- vortex-mixed twice for 1 minute each, and the mixture was
mogenized, total lipids of the kidney and liver homogenates centrifuged at 13,000 g for 10 minutes at 48C to yield a
were extracted with a mixture of chloroform and methanol supernatant containing extracted nuclear proteins for
(2:1, vol=vol) according to the method of Folch et al.,20 and SREBP-1 and -2 protein determination. To ensure equal
the amounts of total cholesterol and triglycerides were de- loading among lanes, the protein concentration of each tis-
termined using the Wako kit as described above. sue was determined using a Bio-Rad protein assay kit and
BSA as a standard, and then immunoblotting was carried
out. For the determination of RAGE, CEL, CML, and
Total and nuclear protein preparations
SREBP-1 and -2, 30 mg of protein from each sample was
and western blot analyses
electrophoresed through 8% or 10% sodium dodecyl sulfate
Each section of the kidney was homogenized with ice- polyacrylamide gels. Separated proteins were electropho-
cold lysis buffer (pH 7.5) containing 137 mM NaCl, 20 mM retically transferred to a nitrocellulose membrane, blocked
Tris-HCl, 1% (vol=vol) Tween 20, 10% (vol=vol) glycerol, with 5% (wt=vol) skim milk solution for 1 hour, and then
1 mM PMSF, and protease inhibitor mixture DMSO solu- incubated with primary antibodies to RAGE, CEL, CML,
tion. After centrifugation (2,000 g at 48C), supernatant of the SREBP-1, SREBP-2, and b-actin, respectively, overnight at
kidney was used for the RAGE, CEL, and CML determi- 48C. After the blots were washed, they were incubated with
nations. A nuclear extract of the liver was obtained ac- goat anti-rabbit and=or goat anti-mouse immunoglobulin G
cording to the method of Rangan et al.21 with minor horseradish peroxidase-conjugated secondary antibody for
modifications. In brief, hepatic tissue (100 mg) was ho- 90 minutes at room temperature. Each antigen–antibody
mogenized in 200 mL of ice-cold hypotonic buffer contain- complex was visualized using ECL western blotting detec-
ing 10 mM HEPES (pH 7.9), 10 mM KCl, 2 mM MgCl2, tion reagents and detected by chemiluminescence with
0.1 mM EDTA, 0.5 mM dithiothreitol, 1 mM PMSF, and LAS-1000 plus (Fujifilm, Tokyo). Band densities were

Table 1. Body Weight, Tissue Weight, and Food and Water Intake in OLETF and LETO Rats
OLETF rats

Matcha (mg=kg of body weight=day)

Control 50 100 200 LETO rats
Body weight (g)
Before 541.5  10.5 543.4  12.6 542.6  7.7 542.1  13.2 435.0  6.8*
4 weeks 582.4  12.5 581.9  15.0 568.7  9.2 564.0  16.0 468.2  8.2*
8 weeks 609.8  12.9 606.6  17.1 585.3  10.7 591.2  17.8 479.7  6.3*
12 weeks 630.3  13.9 621.6  18.4 609.5  11.3 609.1  17.2 497.5  8.0*
16 weeks 642.0  12.0 632.0  21.3 623.3  11.0 613.7  16.0 507.0  9.6*
Tissue weight at 16 weeks (g=100 g of body weight)
Kidney 0.529  0.008 0.537  0.013 0.559  0.010 0.575  0.010* 0.458  0.008*
Liver 3.71  0.07 3.54  0.08 3.43  0.06* 3.39  0.11* 3.03  0.06*
Food intake (g=day)
Before 27.6  0.6 28.4  0.4 26.3  0.6 26.7  1.1 20.7  0.5*
4 weeks 29.8  0.8 29.4  0.7 28.6  0.6 30.5  0.9 21.3  0.3*
8 weeks 29.0  0.6 29.7  0.6 29.4  1.1 29.6  0.9 21.1  0.4*
12 weeks 27.2  0.5 29.2  0.7* 28.1  0.4 29.4  0.4* 20.9  0.4*
16 weeks 27.9  1.0 28.4  2.4 29.3  1.0 28.7  1.2 22.9  1.3
Water intake (mL=day)
Before 35.8  1.4 37.4  1.6 32.1  1.3 34.8  2.0 30.9  1.5
4 weeks 38.6  1.5 41.2  2.0 45.3  3.5 46.7  2.3 32.5  2.5
8 weeks 40.7  1.1 43.5  1.6 44.8  1.4 47.1  1.6* 33.9  0.8*
12 weeks 40.0  1.7 44.4  1.7 49.6  4.1* 50.5  1.3* 32.9  0.6
16 weeks 39.4  1.3 44.9  1.5 47.6  3.2* 48.4  1.8* 33.7  0.9
*P < .05 for significance of difference versus OLETF control rats.
 MATCHA IN TYPE 2 DIABETIC OLETF RATS 717

determined by Scion image software (Scion Corp., Freder- perimental period (16 weeks). All three factors in OLETF
ick, MD) and quantified as the ratio to b-actin. The mean rats were significantly higher than those in LETO rats used
protein levels were indexed to the normal rats with the as a normal group, at the beginning of this experiment. Food
values set at 1, and the corresponding values for the diabetic intake showed a slight increase with the administration of
rats were expressed as the ratios of the normal values. Matcha at 12 weeks, and water intake was significantly in-
creased in a dose-dependent manner after 8 weeks of the
Statistical analysis experimental period. In contrast, the body weight showed a
dose-dependent tendency to decrease in the latter half of this
The results are presented as mean  SE values. The effect
experiment with Matcha administration, but the tendency
of Matcha on each parameter was examined using one-way
was not significant.
analysis of variance. Individual differences among groups
were analyzed by Dunnett’s test, and significance was ac-
cepted at P < .05. Kidney and liver weights
The kidney and liver weights (g=100 g of body weight)
RESULTS were significantly increased in diabetic OLETF compared to
LETO rats (Table 1). The increased liver weights in OLETF
Changes in body weight and food and water intake
rats were significantly decreased by Matcha with 100 and
Table 1 shows the changes in the body weight and food 200 mg=kg treatment, whereas kidney weights were slightly
and water intake in LETO and OLETF rats during the ex- increased by Matcha compared to OLETF control rats.

FIG. 1. Profiles of (A) serum glucose and (B) total cholesterol levels every 4 weeks and (C) urinary protein and (D) Ccr levels every 8 weeks
during 16 weeks of the experimental period: normal (LETO) (&), control (&), Matcha (50 mg=kg) ( ), Matcha (100 mg=kg) ( ), and Matcha
(200 mg=kg) ( ). BW, body weight. #P < .05 for significance of difference versus OLETF control rats.
718 YAMABE ET AL.

Changes in serum glucose, total cholesterol, In addition, the total cholesterol level was markedly reduced
and Ccr levels and urinary protein level in the liver compared with the kidney by Matcha adminis-
tration.
Figure 1 shows the changes in serum glucose and total
cholesterol levels during 16 weeks of the administration
Effects on CML, CEL, and RAGE expression
period. Serum glucose and total cholesterol levels in OLETF
in renal cortex
rats were increased time-dependently, compared with those
of LETO rats. Serum glucose levels in OLETF rats treated To evaluate the effect of Matcha on the expression levels
with Matcha were mildly reduced (Fig. 1A), whereas serum of CML, CEL, and RAGE associated with AGEs in the renal
total cholesterol levels (Fig. 1B) were significantly de- cortex of diabetic OLETF rats, we carried out western blot
creased after 12 weeks in 100 and 200 mg=kg Matcha- analyses. As shown in Figure 2, the expression level of
treated rats. The urinary protein level, which was increased CML, CEL, and RAGE in diabetic OLETF rats was in-
in OLETF rats, was slightly decreased by the administration creased compared with LETO rats (1.46-, 1.35-, and 1.27-
of Matcha at the end of the experiment (Fig. 1C), whereas fold, respectively). However, the elevated expressions of
the Ccr level showed no significant changes during the ex- CML and CEL were greatly reduced by Matcha adminis-
perimental period (Fig. 1D). tration in a dose-dependent manner, whereas RAGE ex-
pression was mildly reduced.
Effects on serum biochemical factors
Effects on SREBP-1 and -2 in hepatic nuclear fraction
As shown in Table 2, the levels of serum glycosylated
protein, triglycerides, and TBARS in diabetic OLETF con- To assess the effects of Matcha on the protein expressions
trol rats significantly increased compared to those of LETO associated with triglyceride and cholesterol metabolism in
rats, whereas serum total protein and albumin levels showed the hepatic tissue of diabetic OLETF rats, we carried out
no difference between the OLETF and LETO animals. The western blot analyses with SREBP-1 and -2 using nuclear
elevated triglyceride and TBARS levels were significantly fractions. As shown in Figure 3, the expression level of
decreased in the OLETF group treated with Matcha at 100 SREBP-1 and -2 in diabetic OLETF rats was decreased
and 200 mg=kg, whereas the serum glycosylated protein compared to that of LETO rats (0.75- and 0.77-fold, re-
level was slightly decreased by Matcha treatment. However, spectively). The decreased volume of SREBP-1 in diabetic
the 200 mg=kg Matcha-treated group showed an increase in OLETF rats was not affected by Matcha treatment. In con-
the serum total protein level compared with the OLETF trast, treatment with 200 mg=kg Matcha elevated the
control group. SREBP-2 expression level to the value of LETO rats.

Effects on glucose, AGE, total cholesterol, DISCUSSION

and triglyceride levels in renal and hepatic tissues
In Japan green tea is classified into several different types,
The levels of glucose, AGEs, total cholesterol, and tri- such as Sencha, Gyokuro, Tamaryokucha, Bancha, Hojicha,
glycerides in both the kidney and liver of diabetic OLETF Matcha, and Kamairicha, according to their constituents and
control rats were significantly elevated compared to LETO manufacturing methods.22 Among them, Matcha is a pow-
rats, except for the liver AGE level (Table 3). Matcha ad- dered green tea produced by grinding with a stone mill after
ministration did not reduce the glucose and triglyceride removing the veins, stems, and impurities from the leaves of
levels in the kidney, but significantly reduced these levels in shade-grown tea trees. Green tea and its major components,
the liver. AGE levels in the kidney were also significantly catechins such as epicatechin, epicatechin 3-O-gallate, epi-
decreased by Matcha administration, but levels in the liver gallocatechin, and epigallocatechin 3-O-gallate, have shown
showed a tendency to decrease (difference not significant). beneficial effects in the treatment and prevention of various

Table 2. Serum Biochemical Levels in OLETF and LETO Rats

OLETF rats

Matcha (mg=kg of body weight=day)

Control 50 100 200 LETO rats
Glycosylated protein (nmol=mg of protein) 19.2  0.4 18.8  0.5 18.4  0.3 18.2  0.4 15.7  0.8*
Albumin (g=dL) 3.42  0.05 3.49  0.03 3.52  0.05 3.55  0.06 3.57  0.02
Total protein (g=dL) 5.60  0.07 5.64  0.07 5.76  0.06 5.86  0.08* 5.55  0.06
Triglyceride (mg=dL) 247.0  19.7 211.3  16.3 190.3  13.8* 188.8  16.6* 65.0  5.4*
TBARS (nmol=mL) 3.05  0.17 2.62  0.20 2.21  0.11* 2.13  0.17* 2.31  0.11*
*P < .05 for significance of difference versus OLETF control rats.
 MATCHA IN TYPE 2 DIABETIC OLETF RATS 719

Table 3. Renal and Hepatic Parameters in OLETF and LETO Rats

OLETF rats

Matcha (mg=kg of body weight=day)

Control 50 100 200 LETO rats
Renal
Glucose (mg=g of wet tissue) 1.43  0.05 1.44  0.04 1.43  0.04 1.47  0.04 1.20  0.04*
AGEs (AU) 0.98  0.03 0.93  0.02 0.88  0.01* 0.88  0.02* 0.85  0.05*
Total cholesterol (mg=g of wet tissue) 4.90  0.07 4.67  0.05* 4.72  0.06* 4.71  0.06* 4.27  0.02*
Triglyceride (mg=g of wet tissue) 6.13  0.12 5.82  0.15 6.14  0.10 6.13  0.21 4.47  0.12*
Hepatic
Glucose (mg=g of wet tissue) 6.95  0.24 6.38  0.21* 6.27  0.11* 6.10  0.18* 5.42  0.27*
AGEs (AU) 0.81  0.02 0.77  0.02 0.76  0.04 0.74  0.03 0.67  0.04
Total cholesterol (mg=g of wet tissue) 7.61  0.26 6.83  0.24 6.60  0.36* 6.14  0.32* 4.58  0.37*
Triglyceride (mg=g of wet tissue) 37.11  1.35 29.49  0.98* 28.90  1.10* 28.68  1.41* 14.25  0.76*
*P < .05 for significance of difference versus OLETF control rats.

human chronic diseases, including cancer, cardiovascular total cholesterol levels were significantly reduced in the
disease, oral disease, arthritis, obesity, diabetes, and Alz- OLETF rats (Fig. 1 and Table 2). In addition, the elevated
heimer’s disease, and also promote longevity through their serum TBARS levels, used as an indicator of oxidative
antioxidant properties.23,24 stress,26 in OLETF rats were also significantly decreased
The control of hyperglycemia and dyslipidemia such as with 100 and 200 mg=kg Matcha administration (Table 2).
hypertriglyceridemia and hypercholesterolemia in type 2 AGEs, formed by the nonenzymatic glycation of sugar
diabetes has been considered a therapeutic strategy along molecules and proteins or lipids, accumulate during the
with body weight loss through lifestyle modification.25 In normal aging process and at accelerated rates during the
the present study, we investigated the effects of Matcha on course of diabetes and are involved in the pathogenesis of
OLETF rats, a model of human type 2 diabetes mellitus, to diabetes and diabetic complications, such as nephropathy,
identify its effects on hyperglycemia, dyslipidemia, AGEs retinopathy, atherosclerosis, and liver diseases, such as non-
and their receptors, and oxidative stress. Matcha (50, 100, or alcoholic steatohepatitis, liver cirrhosis, and cancers, and
200 mg=kg of body weight) was orally administered for 16 chronic diseases, such as arthritis and Alzheimer’s dis-
consecutive weeks to 22-week-old OLETF rats. Following ease.8,12,27 In this experiment, serum glycosylated protein
Matcha administration, serum glucose, triglyceride, and in serum and AGE levels in the liver and kidney were

FIG. 2. Effects of Matcha on (A) CML, (B) CEL, and (C) RAGE levels in the renal cortex. There were no significant differences versus OLETF
control rats.
720 YAMABE ET AL.

FIG. 3. Effects of Matcha on

nuclear (A) SREBP-1 and (B)-2
levels in hepatic tissue. There were
no significant differences versus
OLETF control rats.

increased in OLETF rats. Matcha administration (100 and ment (Table 3). In this experiment, Matcha restored the
200 mg=kg of body weight) greatly reduced the elevated decreased SREBP-2 expression to the values of normal rats,
AGE levels in the kidney, but not in the serum or liver but not that of SREBP-1 (Fig. 3). Currently, AMP-activated
(Tables 2 and 3). Then, we investigated the effects of protein kinase, a key regulator of intracellular energy
Matcha on the expression of RAGE as well as CML and balance, is also considered a therapeutic target for type 2
CEL, two major products of AGEs in the kidney. CML diabetes and metabolic syndrome.34 Epigallocatechin 3-O-
accumulates with TBARS in glomerular lesions, resulting in gallate suppresses hepatic gluconeogenesis through AMP-
structural and functional alterations in plasma and extra- activated protein kinase activation, and green tea causes a
cellular matrix proteins.28 In addition, RAGE is activated by reduction in the level of mRNAs for gluconeogenic en-
CML, and the AGE–RAGE interaction in mesangial and zymes, phosphoenolpyruvate carboxykinase, and glucose
endothelial cells increases reactive oxygen species forma- 6-phosphatase in mouse liver.35,36 Therefore, Matcha may
tion, with the subsequent activation of nuclear factor-kB and also show protective effects against liver damage via a de-
release of pro-inflammatory cytokines such as interleukin-6 crease in the levels of glucose, triglycerides, and total cho-
and tumor necrosis factor-a, growth factors, and adhesion lesterol and an increase in SREBP-2 expression along with
molecules associated with renal disease. Inhibitors and inhibitions of gluconeogenesis through AMP-activated
breakers of AGEs and RAGE blocker may retard the pro- protein kinase activation and gluconeogenic enzyme ex-
gression of diabetic nephropathy or improve the treatment pression.
effects.8,12 Western blotting analyses revealed that Matcha In conclusion, we found that Matcha has beneficial effects
effectively reduced the elevated expression levels of CML, on renal and hepatic damage through the suppression of
CEL, and RAGE in OLETF rats (Fig. 2). Epigallocatechin renal AGE accumulation and reductions in the hepatic
3-O-gallate, a major component of Matcha, also showed glucose, triglyceride, and total cholesterol levels and in
beneficial effects on diabetic renal damage.15 antioxidant activities. Therefore, daily Matcha consumption
SREBPs are transcription factors that regulate fatty acid might have beneficial effects on the control of obesity and
and cholesterol synthesis, as well as insulin sensitivity.29 type 2 diabetes.
SREBP-1 preferentially activates genes involved in fatty
acid synthesis, whereas SREBP-2 preferentially activates
genes involved in cholesterol biosynthesis. SREBP-1 exists AUTHOR DISCLOSURE STATEMENT
as two isoforms (SREBP-1a and SREBP-1c). SREBP-1c No competing financial interests exist.
induces insulin resistance by inhibiting hepatic insulin re-
ceptor substrate 2 signaling,30 which may lead to hyper-
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