Available online at www.sciencedirect.

com

Bioresource Technology 99 (2008) 2981–2988

In vitro studies of eggplant (Solanum melongena) phenolics as

inhibitors of key enzymes relevant for type 2 diabetes and hypertension
Y.-I. Kwon, E. Apostolidis, K. Shetty *

Laboratory of Food Biotechnology, Department of Food Science, Chenoweth Building, University of Massachusetts, Amherst, MA 01003, USA

Received 6 April 2007; received in revised form 5 June 2007; accepted 5 June 2007
Available online 13 August 2007

Abstract

National Diabetes Education Program of NIH, Mayo Clinic and American Diabetes Association recommend eggplant-based diet as a
choice for management of type 2 diabetes. The rationale for this suggestion is the high fiber and low soluble carbohydrate content of
eggplant. We propose that a more physiologically relevant explanation lies in the phenolic-linked antioxidant activity and a-glucosidase
inhibitory potential of eggplant which could reduce hyperglycemia-induced pathogenesis. Results from this study indicate that phenolic-
enriched extracts of eggplant with moderate free radical scavenging-linked antioxidant activity had high a-glucosidase inhibitory activity
and in specific cases moderate to high angiotensin I-converting enzyme (ACE) inhibitory activity. Inhibition of these enzymes provide a
strong biochemical basis for management of type 2 diabetes by controlling glucose absorption and reducing associated hypertension,
respectively. This phenolic antioxidant-enriched dietary strategy also has the potential to reduce hyperglycemia-induced pathogenesis
linked to cellular oxidation stress. These results provide strong rationale for further animal and clinical studies.
 2007 Elsevier Ltd. All rights reserved.

Keywords: Angiotensin I-converting enzyme (ACE); Eggplant phenolics; a-Glucosidase inhibitory activity; Hypertension; Type 2 diabetes

1. Introduction plications of NIDDM due to the hyperglycemia-induced

pathogenesis are metabolic problems and infection (Nis-
Diabetes mellitus (DM) is a major chronic disease hikawa et al., 2000). Hyperglycemia, a condition character-
caused by an improper balance of glucose homeostasis ized by an abnormal postprandial increase of blood glucose
(Rosetti et al., 1990). DM is a serious chronic metabolic level, has been linked to the onset of NIDDM and associ-
disorder that has a significant impact on the health, quality ated oxidation-linked vascular complications (Dicarli et al.,
of life, and life expectancy of patients, as well as on the 2003; Haffner, 1998). Recent studies have shown that the
health care system. At least 171 million people worldwide glucose-induced increased levels of mitochondrial reactive
have diabetes; this figure is likely to be more than double oxygen species (ROS) produced by the mitochondrial
by 2030 and around 3.2 million deaths every year are electron transport chain seems to be the causal link
attributable to complications of diabetes; six deaths every between elevated levels of glucose and the pathways
minute (WHO, 2007). responsible for hyperglycemia-induced vascular complica-
Two types of DM are currently known, one being insu- tions (Kawamura and Heinecke, 1994).
lin-dependent diabetes mellitus, IDDM and the other being Studies indicate that hyperglycemia triggers the genera-
type 2 non-insulin-dependent diabetes mellitus, NIDDM tion of free radicals and oxidative stress in capillary endo-
(Harris and Zimmer, 1992). The most common acute com- thelial cells in the retina, mesangial cells in the renal
glomerulus and neuron cells in the peripheral nerves
*
Corresponding author. Tel.: +1 413 545 1022; fax: +1 413 545 1262. (Brownlee, 2005). Therefore, it is essential to regenerate
E-mail address: [email protected] (K. Shetty). critical cellular antioxidant responses to manage cellular

0960-8524/$ - see front matter  2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2007.06.035
2982 Y.-I. Kwon et al. / Bioresource Technology 99 (2008) 2981–2988

redox status for preventing these diabetic complications MAYOCLINIC, 2007; ADA, 2007). Our hypothesis is that
resulting from hyperglycemia (Heilig et al., 1995; Lee and the biochemical rationale behind this recommendation lies
Chung, 1999). Effective dietary strategies can contribute in the phenolic-enriched antioxidant activity and a-glucosi-
to solutions for managing both hyperglycemia and proper dase inhibitory potential of eggplant, which has the poten-
cellular redox status. tial to reduce hyperglycemia-induced pathogenesis. Any
a-Amylase and a-glucosidase are key enzymes involved dietary management of hyperglycemia linked to type 2 dia-
in starch breakdown and intestinal absorption, respec- betes and related complications from oxidative dysfunction
tively. It is now believed that inhibition of these enzymes can benefit from specific enzyme inhibitory activity com-
involved in the digestion and uptake of carbohydrates bined with antioxidant activity in the same whole food
can significantly decrease the postprandial increase of extracts. This approach has potential for high compliance
blood glucose level after a mixed carbohydrate diet and and less side-effects. Therefore, the objective of this
therefore can be an important strategy in the management research was to evaluate several types of commonly avail-
of hyperglycemia linked to type 2 diabetes (Puls et al., able eggplant varieties for a-amylase, a-glucosidase and
1977). A main drawback of currently used a-glucosidase ACE inhibitory activities using different enzyme sources
and a-amylase inhibitors such as acarbose is side effects such as yeast a-glucosidase, rat intestinal a-glucosidase
such as abdominal distention, flatulence, meteorism and and porcine pancreatic a-amylase and rabbit lung ACE.
possibly diarrhea (Bischoff et al., 1985). It has been sug- These in vitro inhibitory activities were compared to total
gested that such adverse effects might be caused by the phenolic content and antioxidant activity in the water
excessive inhibition of pancreatic a-amylase resulting in extracts of these readily available eggplant varieties.
the abnormal bacterial fermentation of undigested carbo-
hydrates in the colon (Bischoff, 1994; Horii, 1987). There- 2. Methods
fore, natural inhibitors from dietary plants have shown
to have lower inhibitory effect against a-amylase activity 2.1. Extract preparation
and a stronger inhibitory activity against a-glucosidase
and can be used as effective therapy for postprandial hyper- Fresh and well-ripened eggplant (Solanum melongena);
glycemia with minimal side effects (Kwon et al., 2006). Purple, White, Graffiti, Italian were purchased from Big
One of the long-term complications of diabetes is hyper- Y supermarket, Hadley, MA. After peeling of each type
tension, or high blood pressure. Angiotensin I-converting of eggplant with a knife, 15 g of pulp and skin were added
enzyme (ACE) is an important enzyme involved in main- to 10 ml of distilled water separately and homogenized for
taining vascular tension. ACE activates a histidyl-leucine 1 min using a Waring laboratory blender (Winsted, CN) set
dipeptide called angiotensin I, into a potent vasoconstric- on ‘‘HIGH’’. The homogenate was centrifuged (Eppendorf
tor called angiotensin II (Skeggs et al., 1956). Angiotensin 5415D, Brinkmann Instruments Inc., Westbury, NY) at
II also stimulates the synthesis and release of aldosterone, 9300g for 10 min. The supernatant was vacuum filtered
which increases blood pressure by promoting sodium reten- through a Whatman # 2 (Whatman International Ltd.,
tion in the distal tubules (Lieberman, 1975). Inhibition of Maidstone, UK) filter and then used as the crude extract
ACE is considered a useful therapeutic approach in the for in vitro assays.
treatment of high blood pressure in both diabetic and
non-diabetic patients (Erdos and Skidgel, 1987). 2.2. Total phenolics assay
Phenolic compounds or phenolic phytochemicals are
secondary metabolites of plant origin and are important The total phenolics were determined by an assay modi-
parts of the diet (Bravo, 1998; Paganga et al., 1999; Urqui- fied from Shetty et al. (1995). One milliliter of eggplant
aga and Leighton, 2000) providing potential antioxidant extract was transferred into a test tube and mixed with
benefits for managing oxidation stress-related chronic dis- 1 ml of 95% ethanol and 5 ml of distilled water. To each
eases such as diabetes and cardiovascular disease (Serdula sample 0.5 ml of 50% (v/v) Folin–Ciocalteu reagent was
et al., 1996). Vegetables such as eggplant, pepper and added and mixed. After 5 min, 1 ml of 5% Na2CO3 was
tomato of the Solanaceae family have high phenolic con- added to the reaction mixture and allowed to stand for
tent and specifically eggplants are a rich source of phenolic 60 min. The absorbance was read at 725 nm. The absor-
phytochemicals having high free radical scavenging-linked bance values were converted to total phenolics and were
antioxidant activity (Luthria and Mukhopadhyay, 2006). expressed in micrograms equivalents of gallic acid per mil-
Further, understanding the health benefits of eggplant liliter of the sample. Standard curves were established using
has merit when considering the cholesterol-lowering effects various concentrations of gallic acid in 95% ethanol.
of a portfolio diet, which has eggplant as an important
fiber source (Jenkins et al., 2003). The food recipes of the 2.3. Antioxidant activity by 1,1-diphenyl-2-picrylhydrazyl
National Diabetes Education Program of NIH (National radical (DPPH) inhibition assay
Institute of Health), Mayo Clinic and American Diabetes
Association (ADA) recommend eggplant as a part of the To 3 ml of 60 lM DPPH in ethanol, 250 ll of each egg-
diet for management of type 2 diabetes (NIH, 2007; plant extract was added, the decrease in absorbance was
 Y.-I. Kwon et al. / Bioresource Technology 99 (2008) 2981–2988 2983

monitored at 517 nm until a constant reading was was expressed as inhibition % and was calculated as
obtained. The readings were compared with the controls, follows:
which contained 250 ll of 95% ethanol instead of the " #!
extract. The % inhibition was calculated by DAControl
405  DAExtract
% Inhibition ¼  Control 405  100
" #! DA405
AControl
517  AExtract
517
% Inhibition ¼  Control   100
A517
2.6. Angiotensin I-converting enzyme (ACE) inhibition
assay
2.4. a-Amylase inhibition assay
ACE inhibition was assayed by a method modified by
Kwon et al. (2006). The substrate, hippuryl-histidyl-leu-
Porcine pancreatic a-amylase (EC 3.2.1.1) was pur-
cine (HHL) and angiotensin I-converting enzyme (ACE)
chased from Sigma Chemical Co (St. Louis, MO). A total
from rabbit lung (EC 3.4.15.1) were purchased from
of 500 ll of eggplant extract and 500 ll of 0.02 M sodium
Sigma. Fifty microliters of extracts were incubated with
phosphate buffer (pH 6.9 with 0.006 M sodium chloride)
100 ll of 1.0 M NaCl-borate buffer (pH 8.3) containing
containing a-amylase solution (0.5 mg/ml) were incubated
2.0 mU ACE solution at 37 C for 10 min. After pre-incu-
at 25 C for 10 min. After pre-incubation, 500 ll of a 1%
bation, 100 ll of a 5.0 mM substrate (HHL) solution was
starch solution in 0.02 M sodium phosphate buffer (pH
added to reaction mixture. Test solutions were incubated
6.9 with 0.006 M sodium chloride) was added to each tube
at 37 C for 1 h. The reaction was stopped with 150 ll
at 5 s intervals. The reaction mixtures were then incubated
of 0.5 N HCl. The hippuric acid formed was detected
at 25 C for 10 min. The reaction was stopped with 1.0 ml
and quantified by HPLC method. A volume of 5 ll of
of dinitrosalicylic acid color reagent. The test tubes were
sample was injected using Agilent ALS 1100 autosampler
then incubated in a boiling water bath for 5 min and cooled
into an Agilent 1100 series HPLC (Agilent Technologies,
to room temperature. The reaction mixture was then
Palo Alto, CA) equipped with DAD 1100 diode array
diluted after adding 10 ml distilled water and absorbance
detector. The solvents used for gradient were (A) 10 mM
was measured at 540 nm.
phosphoric acid (pH 2.5) and (B) 100% methanol. The
" #! methanol concentration was increased to 60% for the first
AControl
540  AExtract 8 min and to 100% for the 5 min, then decreased to 0% for
% Inhibition ¼  Control540
  100
A540 next 5 min (total run time, 18 min). The analytical column
used was Nucleosil 100-5C18, 250 · 4.6 mm i.d. (Supelco
Inc., Bellefonte, PA), with packing material of 5 lm parti-
2.5. Baker’s yeast and rat intestinal a-Glucosidase cle size at a flow rate 1 ml/min at ambient temperature.
During each run the chromatogram was recorded at
a-Glucosidase (EC 3.2.1.20) was purchased from Sigma 228 nm and integrated using Agilent Chemstation
Chemical Co. A volume of 50 ll of sample solution and enhanced integrator for detection of liberated hippuric
100 ll of 0.1 M phosphate buffer (pH 6.9) containing a-glu- acid. Pure hippuric acid (purchased from Sigma Chemical
cosidase solution (1.0 U/ml) were incubated in 96 well Co., St. Louis, MO) was used to calibrate the standard
plates at 25 C for 10 min. After pre-incubation, 50 ll of curve and retention time. The % inhibition was calculated
5 mM p-nitrophenyl-a-D-glucopyranoside solution in by:
0.1 M phosphate buffer (pH 6.9) was added to each well " #!
at 5 s intervals. The reaction mixtures were incubated at EControl  ESample
25 C for 5 min. % Inhibition ¼  Control   100
E  EBlank
Rat intestine acetone powder as a crude enzyme extract
was purchased from Sigma Chemical Co. A volume of E; efficacy (area of peak, HPLC).
50 ll of sample solution and 100 ll of 0.1 M phosphate
buffer (pH 6.9) containing crude a-glucosidase solution 2.7. Statistical analysis
(25 mg/ml) were incubated in 96 well plates at 25 C for
10 min. After pre-incubation, 50 ll of 5 mM p-nitro- All experiments were performed at least in duplicates.
phenyl-a-D-glucopyranoside solution in 0.1 M phosphate Analysis at every time point from each experiment was car-
buffer (pH 6.9) was added to each well at 5 s intervals. ried out in triplicates. Means, standard errors, standard
The reaction mixtures were incubated at 37 C for 30 min. deviations and Pearson’s correlation coefficient were calcu-
In both cases before and after incubation, absorbance lated from replicates with in the experiments and analyses
readings were recorded at 405 nm by micro-array reader were done using Microsoft Excel 2003. The results were
(Thermomax, Molecular device Co., Sunnyvale, CA) and statistically analyzed by ANOVA and Duncan’s multiple
compared to a control which had 50 ll of buffer solution range tests. Statistical significance was accepted at a level
in place of the extract. The a-glucosidase inhibitory activity of p < 0.05.
2984 Y.-I. Kwon et al. / Bioresource Technology 99 (2008) 2981–2988

3. Results tory activity with water extract and 12% ethanol extracts
(Kwon et al., 2006). In order to have better health rele-
3.1. Total phenolics and antioxidant activity vance mammalian a-glucosidase (rat intestine) and yeast
a-glucosidase (baker’s yeast) inhibitory activities of pulp
The skin extract of Italian, which had 94.7 ± 13.3 lg/ml and skin extracts of eggplant were compared. Except for
of total soluble phenolics and was highest among all the Italian variety, the microbial a-glucosidase inhibitory activ-
eggplant extracts evaluated (Fig. 1). The pulp extract of ity of pulp and skin extracts of eggplant was directly pro-
Graffiti, pulp extract of White and skin extract of Purple portional to the mammalian a-glucosidase inhibitory
eggplant had 59.7 ± 0.9 lg/ml, 56.7 ± 5.1 lg/ml and activity (Fig. 2; Pearson’s correlation coefficient was
44.7 ± 12.3 lg/ml of soluble phenolics, respectively (Fig. 1). 0.8627 at p < 0.05). Among various extracts, pulp of White
The antioxidant activity of the extracts was monitored and Graffiti had high a-glucosidase inhibitory activity
using the DPPH radical inhibition (DRI) assay. The ability using both rat intestine and yeast sources. In others Purple
of phenolics in eggplant extracts to inhibit the DPPH radi- pulp and Graffiti skin showed moderately high inhibitory
cal formation was measured. The skin extract of Italian egg- activity using the more relevant rat source and much higher
plant had the highest DPPH radical inhibition (DRI) inhibitory activity using the yeast source. Except for Italian
capacity (46%) followed by pulp extract of White (35%), eggplant, both microbial a-glucosidase inhibitory activity
pulp extract of Graffiti (34%) and pulp extract of Purple and mammalian a-glucosidase inhibitory activity of pulp
(33%) (Fig. 1). Except for Italian eggplant, the DRI capac- and skin extract of eggplant were also proportional to
ity of all pulp extract samples was higher than skin extracts. the antioxidant activity (Figs. 1 and 2; Pearson’s correla-
Except for purple eggplant, DPPH radical inhibition activ- tion coefficient, 0.7893 and 0.6803 at p < 0.05, respectively).
ity of all extract samples was proportional to the concentra- Analysis also indicated a good dose dependent response
tion of total soluble phenolics (Fig. 1; Pearson’s correlation (data not shown).
coefficient was 0.8549 at p < 0.05). These results suggest The a-amylase inhibitory activity was highest in Graffiti
that higher phenolic content does confer moderate antioxi- pulp and much lower in other samples (Fig. 3). Previous
dant activity linked to free radical scavenging potential. research with select food extracts reported an association
between antioxidant activity and a-amylase inhibition
3.2. Glucosidase/amylase inhibition activity (McCue et al., 2005) and therefore the rationale
for this study. The antioxidant activity (DPPH scavenging
Previous research with clonal herbal extracts only activity), mammalian a-glucosidase (rat intestine) and a-
reported a microbial (baker’s yeast) a-glucosidase inhibi- amylase (porcine) inhibitory activities with various water

200 50
Total Phenolics
a

DPPH
b 40
DPPH Scavenging Activity (%)

b
Total Phenolic content (μg/ml)

150
bc
A
30

100 d
d
d
20
BC B
BC BC
CD BC
50
e 10

0 0
Purple Purple White Pulp White Skin Graffiti Graffiti Italian Italian
Pulp Skin Pulp Skin Pulp Skin

Fig. 1. Total soluble phenolics and DPPH radical scavenging activity in water extract of pulp and skin of Eggplant. Total soluble phenolic content (lg/ml)
is plotted with DPPH radical scavenging activity (%). The extraction method is explained in Section 2. A–DValues are the means ± SD of total soluble
phenolic content of three replicated samples. a–eValues are the means ± SD of DPPH radical scavenging activity of three replicated samples. Bar with
different letters indicate statistically significant differences among groups at p < 0.05. Except for purple eggplant, the Pearson’s correlation coefficient
between total phenolic contents and DPPH scavenging activities is 0.8549.
 Y.-I. Kwon et al. / Bioresource Technology 99 (2008) 2981–2988 2985

70
Glucosidase from Rat intestine

a Glucosidase from Yeast

60
b b
c
50 A A d
d e
e B
% Inhibition

40 C
D
D
30

20
E

10
E

0
Purple Purple White White Skin Graffiti Graffiti Italian Italian
Pulp Skin Pulp Pulp Skin Pulp Skin

Fig. 2. The comparison of rat intestinal and yeast a-glucosidase inhibitory activity of water extract of eggplant. Each assay carried out with water extracts
of eggplant. A–EValues are the means ± SD of rat intestinal a-glucosidase inhibitory activity of three replicated samples. a–eValues are the means ± SD of
yeast a-glucosidase inhibitory activity of three replicated samples. Bar with different letters indicate statistically significant differences among groups at
p < 0.05. The Pearson’s correlation coefficient except for Italian eggplant between yeast a-glucosidase and rat intestinal a-glucosidase inhibitory activities
is 0.8627.

extracts of eggplant were compared. Except for skin extract content and antioxidant activity (Figs. 1 and 4A; Pearson’s
of Italian, the a-amylase and a-glucosidase inhibitory correlation coefficient was 0.1416 at p < 0.05). In the dose
activities of skin and pulp of eggplant were proportional dependent study the pulp extract of White specifically indi-
to the DPPH scavenging-linked antioxidant activity cated significantly higher ACE inhibitory activity com-
(Fig. 3; Pearson’s correlation coefficient, 0.7067 and pared to White skin and Graffiti pulp (Fig. 4B).
0.6803 at p < 0.05, respectively).
Although the selected eggplant extracts showed moder-
4. Discussion
ate a-amylase inhibitory activities, the same extracts
(except for Italian pulp and skin) had high a-glucosidase
Hyperglycemia, a condition characterized by an abnor-
inhibitory activity (Fig. 3). When comparing all samples
mal postprandial increase of blood glucose level, has been
White pulp, Graffiti pulp and Purple pulp had high a-glu-
linked to the onset of type 2, non-insulin-dependent diabe-
cosidase, moderate to low a-amylase inhibitory activities
tes mellitus and associated vascular complications (Dicarli
combined with moderate antioxidant activity (Fig. 3).
et al., 2003; Haffner, 1998). Recent studies have indicated
that hyperglycemia-induced vascular complications are
3.3. ACE inhibition likely from oxidative dysfunction from reactive oxygen spe-
cies (ROS) produced by the mitochondrial electron trans-
Inhibition of angiotensin I-converting enzyme (ACE) by port chain (Kawamura and Heinecke, 1994; Brownlee,
dietary anti-hypertensive agents is potentially an important 2005). This ROS dysfunction is linked between elevated
strategy to manage hypertension. Previous research with levels of glucose and the pathways responsible for hyper-
clonal herbal extracts reported ACE inhibitory activity glycemia-induced vascular complications (Kawamura and
combined with a-glucosidase inhibitory activity (Kwon Heinecke, 1994).
et al., 2006) and therefore included in this study. The rabbit Recent studies in rats have suggested that anthocyanin
lung ACE inhibitory activities with water extract of egg- and its derivatives such as nasunin have antioxidant activ-
plant were compared. All samples were prepared on con- ity under oxidative stress (Kimura et al., 1999; Tsuda,
stant volume basis (50 ll). 2000) and caffeolsophorose, diacylated anthocyanin from
Among various eggplant samples, pulp extract of White sweet potato has anti-hyperglycemia effect in rat study
had the highest ACE inhibitory activity (71%) followed by (Matsui et al., 2004). In an in vitro study calystegines from
skin extract of White (48%) and pulp extract of Graffiti eggplant, a type of tropane alkaloids has glycosidases
(38%) (Fig. 4A). Generally ACE inhibitory activity of the inhibitory activity (Asano et al., 1997). Based on these pre-
extracts did not correlate with the total soluble phenolic vious results further phenolic-linked biochemical rationale
2986 Y.-I. Kwon et al. / Bioresource Technology 99 (2008) 2981–2988

60 70
Rat intestinal Glucosidase
Porcine pancreatic Amylase

A DPPH Scavenging Activity 60

50
A
B
a 50
40
C

% DPPH Scavenging Activity

D
b bc 40
% Inhibition

D
30
bc cd bcd
30
cd

20
d 20
E

10
10
E

0 0
Purple Purple White White Graffiti Graffiti Italian Italian
pulp Skin pulp Skin pulp Skin pulp Skin

Fig. 3. The comparison of antioxidant activity, rat intestinal a-glucosidase and porcine a-amylase inhibitory activity of water extract of eggplant. Each
assay carried out with water extracts of eggplant. Each value is the mean of triplicates standard error. A–EValues are the means ± SD of rat intestinal a-
glucosidase inhibitory activity of three replicated samples. a–dValues are the means ± SD of porcine pancreatic a-amylase inhibitory activity of three
replicated samples. Bar with different letters indicate statistically significant differences among groups at p < 0.05. The Pearson’s correlation coefficient
between DPPH scavenging activity and two a-glucosidases (rat intestinal a-glucosidase and porcine a-amylase) are 0.6803 and 0.7067, respectively.

for anti-hyperglycemia effect of eggplants was investigated. will be more preferable. Combinations of enzyme inhibi-
The in vitro results from this research indicate that eggplant tory activity found in eggplant samples White pulp and
varieties enriched in phenolic phytochemicals and moder- Graffiti skin have interesting functionality for potentially
ate free radical scavenging-linked antioxidant activity have controlling glucose absorption with high a-glucosidase
the potential to reduce hyperglycemia-induced vascular inhibition and not generating undigested starch linked
complications resulting from oxidative damage. Most phe- side-effects with low a-amylase inhibition. Undigested
nolic phytochemicals that have positive effect on health are starch linked complications from drug therapy results in
believed to be functioning by countering the effects of ROS the abnormal bacterial fermentation of undigested carbo-
generated during cellular energy metabolism (Bravo, 1998; hydrates in the colon (Bischoff, 1994; Horii, 1987) and
Urquiaga and Leighton, 2000; Serdula et al., 1996). therefore dietary plant-based solution has the potential to
The results from this in vitro study provide the biochem- overcome this challenge.
ical rationale that phenolic-linked ingredients of eggplant Further benefit from this study is the indication that
have the potential for intestinal a-glucosidase inhibition. some specific types of eggplant have potential to inhibit
This indicates the potential to reduce glucose absorption ACE in vitro, which indicates potential anti-hypertension
in the intestine. Additionally several eggplant samples activity. This could provide additional benefit coupled to
(e.g., White pulp and Graffiti skin) have high a-glucosidase cholesterol-lowering effects of portfolio diet, which has egg-
inhibitory activity combined with low a-amylase inhibitory plant as an important fiber source (Jenkins et al., 2003).
activity. Further, the eggplant types investigated have dif- ACE inhibitory activity of the eggplant extracts did not
ferent combinations of in vitro enzyme inhibitory selectivity correlate with the total soluble phenolic content and anti-
and these different responses to microbial and mammalian oxidant activity and may be potentially linked to other
enzyme sources provides sound biochemical rationale for unknown factors. Among all eggplant type evaluated
designing in vivo study using animal model such as rat White (skin and pulp) and Graffiti pulp combine high to
and mouse, where mammalian in vitro enzyme response moderate ACE inhibitory activity with moderate antioxi-
 Y.-I. Kwon et al. / Bioresource Technology 99 (2008) 2981–2988 2987

100 dysfunction-linked diabetic complications can also be

a
90 designed in part through the same food systems.
80 Therefore, results from this study provide a strong bio-
A chemical rationale for the use of eggplant diet as one of the
% Inihibition

70
choices for improving condition of patients with type 2 dia-
60
betes and have been recommended by National Diabetes
50 B
Education Program of NIH (NIH, 2007), American Diabe-
40 C
tes Association (ADA, 2007) and Healthy Recipes of Mayo
30 Clinic (MAYOCLINIC, 2007). Based on this in vitro study
20 further animal and clinical studies can be pursued.
10
D D D D D
0
Purple Purple White White Graffiti Graffiti Italian Italian References
pulp Skin pulp Skin pulp Skin pulp Skin
American Diabetes Association, (ADA). <http://vgs.diabetes.org/
100
b 90
recipe/viewRecipeDisplay.jsp?SizedRecipeId=758&CategoryID=11>
10 ul (accessed March, 2007).
25 ul
80 50 ul Asano, N., Kato, A., Matsui, K., Watson, A.A., Nash, R.J., Molyneux,
% Inhibition

70 R.J., Hackett, L., Toppong, J., Winchester, B., 1997. The effects of
calystegines isolated from edible fruits and vegetables on mammalian
60
liver glycosidases. Glycobiology 7, 1085–1088.
50 Bischoff, H., Puls, W., Krause, H.P., Schutt, H., Thomas, G., 1985.
40 Pharmacological properties of the novel glucosidase inhibitors BAY m
30 1099 (miglitol) and BAY o 1248. Diabetes Res. Clin. Pract. 1, 53–62.
Bischoff, H., 1994. Pharmacology of glucosidase inhibitor. Eur. J. Clin.
20
Invest. 24, 3–10.
10 Bravo, L., 1998. Phenolic phytochemicals: chemistry, dietary sources,
0 metabolism, and nutritional significance. Nutr. Rev. 41, 317–333.
White pulp White Skin Graffiti pulp Brownlee, M., 2005. The pathobiology of diabetic complications. Diabetes
54, 1615–1625.
Fig. 4. Rabbit lung angiotensin I-converting enzyme (ACE) inhibitory Dicarli, M.F., Janises, J., Grunberger, J., Ager, J., 2003. Role of chronic
activity of eggplant (a) and dose dependent changes in ACE inhibitory hyperglycemia in the pathogenesis of coronary microvascular dysfunc-
activity of water extract of eggplant (ll, sample) (b). White (pulp and skin) tion in diabetes. J. Am. Coll. Cardiol. 41, 1387–1393.
and Graffiti pulp combine moderate to high ACE inhibitory activity. Erdos, E.G., Skidgel, R.A., 1987. The angiotensin I-converting enzyme.
A–D
Values are the means ± SD of ACE inhibitory activity of three Lab. Invest. 56, 345–348.
replicated samples. Bar with different letters indicate statistically signif- Haffner, S.M., 1998. The importance of hyperglycemia in the nonfasting
icant differences among groups at p < 0.05. state to the development of cardiovascular disease. Endocrine Rev. 19,
583–592.
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a-amylase inhibitory activity. These varieties therefore other categories of glucose intolerance. In: Alberti, K.G.G.M., De
Fronso, R.A., Keen, H., Zimmer, P. (Eds.), International Textbook of
have best potential for further in vivo studies.
Diabetes Mellitus. John Wiley, London, UK, pp. 3–18.
Heilig, C.W., Concepcion, L.A., Riser, B.L., Freytag, S.O., Zhu, M.,
Cortes, P., 1995. Overexpression of glucose transporters in rat
5. Conclusion
mesangial cells cultured in a normal glucose milieu mimics the diabetic
phenotype. J. Clin. Invest. 96, 1802–1814.
Although this is an in vitro study and the sample size Horii, S., 1987. Synthesis and a-D-glucosidase inhibitory activity of N-
(varieties) evaluated is not extensive, the results indicate substituted valiolamine derivatives as potent oral antidiabetic agents.
the positive effect of the two eggplant types, such as White J. Med. Chem. 29, 1038–1046.
and Graffiti on hyperglycemia risk factors and the bio- Jenkins, D.J., Kendall, C.W., Marchie, A., Faulkner, D.A., Wong, J.M.,
De Souza, R., Emma, A., Parker, T.L., Vidgen, E., Lapsley, K.G.,
marker of hypertension (ACE). It is clear from these Trautwein, E.A., Josse, R.G., Leiter, L.A., Connelly, P.W., 2003.
in vitro assays that White and Graffiti eggplant types have Effects of a dietary vs. lovastatin on serum lipid ands C-reactive
moderate antioxidant activity and good inhibitory profile protein. JAMA 290, 502–510.
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sidase related to glucose absorption in the intestine. Fur- of glucose promote oxidative modification of low density lipoprotein
by a superoxide dependent pathway. Chait. J. Clin. Invest. 94, 771–
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