Journal of Life Science 2009 Vol. 19. No. 1.

40~45 ⓒJLS / ISSN 1225-9918

Antidiabetic Effects of Leaves Extracts of Psidium guajava L. and Lagerstroemia

speciosa L. in STZ-induced Rats
Sang Geun Roh, Kyun Ha Kim and Won Chul Choi*
Department of Biology, College of Natural Sciences, Pusan National
University Received December 4, 2008 /Accepted January 21, 2009

Guava (Psidium guajava L.) and banaba (Lagerstroemia speciosa L.) are well known as medicinal
plants for their antidiabetic effects. These contain a great deal of polyphenol compound and work
on the treatment of diabetes mellitus effectively. In this study, the extracts of guava and banaba are
con-sumed by streptozotocin (STZ) induced diabetic rats to compare the antidiabetic effects.
According to the comparison result, the glucose level of those STZ-induced diabetic rats has
decreased by 19-32%, total cholesterol by 24-46%, triglyceride by 22-67% and free fatty acid by 49-
71% approximately com-pared to the diabetic rats, while the generation of insulin and the recovery
of beta cells have increased. However, the result showed that the antidiabetic effect of guava
extracts was higher than that of banaba extracts. This is because the hydrophilic polyphenol
compounds contained in banaba leaves were not extracted during the ethanol extraction process,
and the antidiabetic activity of the extracted corosolic acid was low to surprise.

Key words : Guava (Psidium guajava), banaba (Lagerstroemia speciosa), polyphenol, anti-diabetic effects

Introduction pounds may differ depending on extraction methods. If the

ethanol extraction method is used, polyphenol compounds
Diabetes mellitus (DM) causes the chronical can hardly extracted.
metabolism disorder which makes approx. 4% of the Psidium guajava L. known as guava is a representative med-
world population suffer and this figure is expected to ical plant that has antidiabetic effects and the interest on gua-va
increase to 5.4% [16]. Hyperglycemia triggered by DM is has been increased recently. Guava leaves contain a variety of
usually caused due to the decrease in insulin secretion, polyphenol compounds such as polyphenol [25], terpe-noids
decrease in glucose uti-lization of tissue and increase in [3,23], flavonoids [20] and tannins [29]. And it was re-ported that
the generation of glucose. Hyperglycemia's symptoms guava leaves have antidiabetic effects direct and indirect through
include obsessive thirst, gluco-suria, polyuria, lipemia and controlling glucose increase [22] and carbo-hydrate hydrolyzing
hunger, and if left without treat-ment, this can result in enzyme's activity [21]. Lagerstroemia spe-ciosa L. known as
patient's death due to critical ketoaci-dosis [27]. banaba has been used in treating DM and kidney diseases for
For ages, many countries have applied traditional medical ages. The antidiabetic effects of banaba was reported by Garcia
plants to treat hyperglycemia and diabetes mellitus [1,6,13,26], in 1940 for the first time, and its ef-fects in the glucose
and more than 400 plants have been reported with their glu-cose decreasing and insulin-like activity were proved through further
reduction effects [2]. Recently the interest on medical plants is researches [9-12]. Murakami and workers (1993) proposed that
getting high as oral agents used to treat DM cause various side corosolic acid is the most effec-tive substance among banaba
effects [4,15] while taking traditional medical plants in a natural leaves's antidiabetic effects [24] and banaba leaves were used
form of extracts show treatment effect as good as that of widely to treat DM. In this vein, the ethanol-extract of banaba
therapeutic agents without side effects. leaves and water-extract of gua-va leaves were consumed by
In general, traditional medical plants contain a wide range streptozotocin (STZ) induced diabetic rats and their antidiabetic
of polyphenol compounds such as flavonoids, glycosides effects were compared.
and terpenoids, which are effective in treating DM [5,7,31].
However, the amount and efficacy of polyphenol com- Materials and Methods

*Corresponding author
Preparation of the plant extract
*Tel：+82-51-510-2262, Fax：+82-51-510-3736
*E-mail : [email protected] The banaba (Lagerstroemia speciosa) leaves extract was pur-
 Journal of Life Science 2009, Vol. 19. No. 1 41

chased from Japan (Wellness banaba Inc, Korea) which was as follows. NC served as normal controls and received ve-
extracted containing 1% of corosolic acid by 90% ethanol hicle only, DC served as diabetic controls and received
extraction. As for guava (Psidium guajava) leaves extract, vehicle. PG received the guava water extract, LS received
150 kg of well dried guava leaves were purchased from the banaba ethanol extract. Each group was composed of 10
Daegu Yangyeongsi Herb Market (Daegu, Korea) and 10 rats. The weight of rats per group was measured daily for 4
o
times of DW was added. This was boiled for 6 hr at 98 C to wk. Guava and banaba extracts were orally ad-ministered by
get extracts. This plant water-soluble extract was filtered with rats twice a day based on the ratio of 1.5 ml per 1 kg of
a 50 μm filter and concentrated under vacuum and freeze- weight. In this case, the same amount of normal saline was
dried to powder form. taken by diabetic rats.

Animal Measuring glucose, insulin and lipid concentration

Six-week-old male Wistar Hannover rats (n=40) were pur- The glucose concentration of the animals was measured
chased from Samtako Experiment Animal (Samtako Bio, once a week. To measure the glucose concentration, all the
Kyung Gi-Do, Korea) and they were acclimatized under a animals were not fed for 8 hr and then blood was sampled
o
controlled temperature of 20±2 C, humidity of 55±5%, 12h from the vein in the tail. The levels of insulin, total cholester-
light and 12 hr dark cycle conditions for 1 week before start- ol (TC), HDL-cholesterol (HDL-C), triglyceride (TG) and free
ing the experiments. They had free access to standard fatty acid (FFA) were measured in week 4. After leaving ani-
pellets (Samtako Bio, Korea) and water. mals without feeding for 8 hr, they were anesthetized with
ethyl ether and then the blood from the heart was sampled
Reagent and equipment and analyzed.
The glucose concentration of animals was measured using
ACCU-CHEK Active (Roche, Germany) and the lipid concen- Histological examination
tration was analyzed using ADVIA 1650 (Bayer, Deerfield, IL, After sampling blood from the animals in week 4, the
USA). The insulin concentration was measured with Sunrise pancreas was separated. Then, the pancreas was fixed in
Remote Control (TECAN, Salzburg, Austria) using Rats Insulin 10% neutral formalin for 1 day and processed in a routine
ELISA Kit (Shibayagi, Japan). Pancreas was cut using Rotary man-ner to 4 μm thick paraffin sections. Hematoxylin-eosin
microtome (Leica, Wetzlar, Germany) and em-ployed (HE) stain was undertaken for the pancreas tissues and its
Hematoxylin-eosin. These preparations were observed under a histo-logical damage was scored as follows based on the
light microscope (Carl Zeiss, Göttingen, Germany). Tang et al., method [30]. 0: normal; Ⅰ: minor injury; Ⅱ:
moderate injury; Ⅲ: obvious injury; Ⅳ: severe injury. Each
Causing diabetics sample was observed at 200x magnification. The degree of
To cause diabetics, streptozotocin (STZ) (65 mg/kg) was injuries of the islet cells was expressed as the mean of 10
solved into the 0.01M citrate buffer (pH 4.5) and then 250 different fields in each slide.
μl was injected with single I.P. injection to rats who were

not fed for 24 hr. Three days after injecting STZ, we meas- Statistical analysis
ured the level of glucose using the blood from the vein in the
All values are expressed as means±S.D. The data
tail. Those rats with blood glucose level of 300 mg/dl or were analyzed by Mann-Whitney test and Kruskal-
higher were regarded as diabetic rats. For normal control Wallis test with SPSS WIN, ver. 12.0. The level of
rats, the same amount of 0.01M citrate buffer was injected statistical significance in the study was p<0.05.
(I.P. injection). Blood sugar levels were measured using
ACCU-CHEK Active (Roche, Germany), and those rats who Results
exceeded 600 mg/dl, which is the limit of glucose measuring
range of the device were excluded in the experiments. Effect of plant extracts on Langerhans islets'
morphology
Grouping animals and feeding extracts Fig. 1 shows the result of HE stain for the pancreas. The
Rats with similar in weight were selected and classified shape of DC's Langerhans islets is smaller and shrunken
42 생명과학회지 2009, Vol. 19. No. 1
Fig. 2 shows the change in the animals' glucose concentration

Fig. 1. Effect of guava and banaba extracts on the histological

micrograph of rats Langerhans islets cell. (HE 200×).
Abbreviations: A, Normal control rats; B, Diabetic con-trol
rats; C, Diabetic rats fed with guava leaves extract; D,
diabetic rats fed with banaba leaves extract. A showed a
normal shape of Langerhans islets cell; B have severely
injured Langerhans islets cell; C, D showed recovery of
Langerhans islets cell.

Table 1. Effect of guava and banaba extracts on the histopatho-

logical damage of pancreas in diabetic rats (n=10)

Pathological grading of pancreas

Group P
0 Ⅰ Ⅱ Ⅲ Ⅳ

NC 10 0 0 0 0 -
DC 0 0 1 4 5 0.000*
PG 0 1 3 3 3 0.000*
*
LS 0 1 2 3 4 0.000
*
P<0.01 are calculated by Kruskal-Wallis’ test and
compared with normal control group.
Abbreviations: NC, Normal control; DC, Diabetic control;
PG, Diabetic mice fed with guava leaves extract; LS,
Diabetic mice fed with banaba leaves extract.

compared to that of NC. Furthermore, in the pathological grading

of pancreas (Table. 1), NC's Langerhans islets were all normal
while DC's were injured mostly (p<0.01). PG's and LS's
Langerhans islets were recovered in terms of shape and size
compared to DC's. PG's injured Langerhans islets were more
recovered than LS's. In the pathological grading of pan-creas,
the number of injured PG's and LS's Langerhans islets was
smaller than DC's, although there was no significant dif-ference
between the two groups in terms of statistics.

Effect of plant extracts on glucose, insulin and lipid

concentration
Fig. 2. Effect of guava and banaba extracts on blood glucose
level in diabetic rats (n=10; mean±S.D.). P<0.01,
com-pared with DC; P<0.01, PG compared with LS.
Abbreviations: NC, Normal control; DC, Diabetic con-
trol; PG, Diabetic mice fed with guava leaves extract;
LS, Diabetic mice fed with banaba leaves extract.

measured in week 4. NC's glucose concentration was low as

86-88 mg/dl for 4 weeks while DC's glucose concen-tration
increased gradually up to higher than 500 mg/dl in week 4.
PG's and LS's glucose concentration started decreas-ing
from week 3 compared to DC's (p<0.01), and PG's glu-cose
concentration was lower than LS's (p<0.01).
Fig. 3 shows the animals' insulin concentration measured
in week 4. NC's insulin concentration was higher as much as
752.2 pg/ml while DC's decreased by approx. 8% of NC's
(p<0.01). Both PG's and LS's insulin concentration increased
more than two times compared to DC's, and PG's insulin
concentration was higher than LS's (p<0.01).
Fig. 4 - Fig. 6 show the animals' lipid concentration
measured in week 4. DC's total cholesterol concentration
was more two times higher than NC's, and the concen-tration
of triglyceride and free fatty acid increased by more than 4
times (p<0.01). The concentration levels of PG's and

Fig. 3. Effect of guava and banaba extracts on serum

insulin level in diabetic rats (n=10; mean±S.D.).
P<0.01, com-pared with DC; P<0.01, PG
compared with LS. NC, DC, PG, LS See Fig. 2.
 Journal of Life Science 2009, Vol. 19. No. 1 43

to the interaction of the genetic, environmental and

immune factors, which results in the destruction of beta
cell in the pancreas [27]. As the insulin secretion
decreases, Type 1 DM develops hyperglycemia and
hyperlipidemia, and if hyper-glycemia is further
developed, this can result in various complications. Thus,
increasing insulin secretion and re-ducing hyperglycemia
is very important in treating type 1 DM patients.
In this experiment, the glucose concentration of diabetic
Fig. 4. Effect of guava and banaba extracts on total control rats was 500 mg/dl or higher, and the insulin con-
cholesterol concentration in diabetic rats (n=10; centration was reduced by 8% of the normal control rats. In
mean±S.D.). P<0.01, compared with DC; P<0.01,
addition, the lipid concentration of diabetic control rats increased
PG compared with LS. NC, DC, PG, LS See Fig. 2.
2 to 4 times higher than that of normal control rats, which
showed the characteristics of Type 1 DM (hyperglycemia,
hyperlipidemia, and hypoinsulinemia). However, consuming
guava and banaba leaves extracts re-duced the glucose
concentration level of diabetic rats by 19% or higher, and
increased the insulin secretion by more than 2 times. These
extracts had antidiabetic effects by reducing the developed
hyperlipidemia and recovering the damaged beta cell of lgerhans
islets. However, there was difference in antidiabetic effects of the
extracts used in this experiment as consuming guava extracts
Fig. 5. Effect of guava and banaba extracts on was more effective compared to consuming banaba extracts in
triglyceride con-centration in diabetic rats (n=10; increasing insulin secretion, reducing glucose level, improving
mean±S.D.). P<0.01, compared with DC; P<0.01, hyperlipidemia and re-covering the beta cell of Langerhans
PG compared with LS. NC, DC, PG, LS See Fig. 2.
islets. In general, poly-phenols are contained in the seeds, fruits,
leaves and barks of medicinal plants [28] and protect plants from
other ex-ternal invaders [26]. Therefore, the antidiabetic effects
of medical plants can be triggered by various flavonoids, glyco-
sides and terpenoids [5,7,31]. The amount of polyphenol may
vary depending on the extraction method as the more ethanol
we use for extraction, the less hydrophilic poly-phenol
compounds we can extract. Actually, in the previous experiment
that used different extract solvents [17], the amount of extracted
polyphenol of banaba leaves extracts was as low as half of the

Fig. 6. Effect of guava and banaba extracts on free fatty acid current experiment since it used 90% ethanol only. So far, it is
concentration in diabetic rats (n=10; mean±S.D.). known that the major anti-diabetic compound contained in
P<0.01, compared with DC; P<0.01, PG compared banaba leaves is corosolic acid [8,18,24], and this substance is
with LS. NC, DC, PG, LS See Fig. 2.
hydrophobic material which is hardly soluble in water. The
solvents with higher ethanol ratio has been used to extract
LS's total cholesterol, triglyceride and free fatty acid were
corosolic acid during the extraction process of banaba leaves,
lower than DC's (p<0.01), and PG's was lower than LS's
and the ethanol ex-tracted banaba leaves that contain 1% of
when comparing just PG's and LS's (p<0.01).
corosolic acid has been commercialized. However, other
researchers [14,19] propose that the antidiabetic compound of
Discussion
these extracts is

Type 1 diabetes mellitus (DM) is a disease caused due

44 생명과학회지 2009, Vol. 19. No. 1

not corosolic acid which is hydrophobic material, but anoth- Novel lipid-lowering properties of Vaccinium myrtillus
er polyphenol which is hydrophilic material as the water L. leaves, a traditional antidiabetic treatment, in
several mod-els of rats dyslipidaemia: a comparison
extracted banaba extracts that rarely contain corosolic acid
with ciprofibrate. Thrombosis Research 84, 311-322.
showed higher antidiabetic effects. On the other hand, guava
7. Farkes, L. 1980. Active principles of plants of
water extracts are known to have antidiabetic effects such as traditional medicine as models of new drugs. Journal of
decreasing plasma sugar level and restraining the activity of Ethnopharmacol-ogy 2, 45-48.
carbohydrate hydrolyzing enzyme [21,22]. Thus, the rea-son 8. Fukushima, M., F. Matsuyama, N. Ueda, K. Egawa, J.
Takemoto, Y. Kajimoto, N. Yonaha, T. Miura, T. Kaneko,
that the antidiabetic effects of banaba ethanol extracts were
Y. Nishi, R. Mitsui, Y. Fujita, Y. Yamada and Y. Seino. 2006.
lower than those of guava water extracts is because the Effect of corosolic acid on postchallenge plasma glucose levels.
majority of hydrophilic polyphenol compounds con-tained in Diabetes Research and Clinical Practice 73, 174-177.
banaba leaves were not extracted during the ex-traction 9. Garcia, F. 1940. On the hypoglycemic effect of decoction of
Lagerstroemia speciosa leaves (banaba) administered orally. Journal
process with ethanol, and the extracted corosolic acid had
of the Philippine Medicinal Associations 20, 395-402.
low antidiabetic activity compared to what we expected.
10. Garcia, F. 1955. Plantisul compared with insulin. Journal
In conclusion, guava water extracts showed higher anti- of the Philippine Medicinal Associations 31, 276-282.
diabetic effects on STZ induced rats compared to banaba ex- 11. Garcia, F. 1956, Plantisul tablets in the treatment of
tracts that contain 1% of corosolic acid. This is because the diabetes mellitus. Journal of the Philippine Medicinal
Associations 31, 216-224.
corosolic acid contained in banaba extracts has low anti-
12. Garcia, F. and P. Melencio-Maglalang. 1957.
diabetic activity and less hydrophilic polyphenol com-pounds Application of banabins (a plantisul preparation) and S.B.
which have high antidiabetic effects. Therefore, fur-ther study menus to diabetics. Journal of the Philippine Medicinal
should be carried out to reveal what kind of hy-drophilic Associations 33, 7-15.
13. Gori, M. and R. K. Campbell. 1998. Natural products and
polyphenol has high antidiabetic effects.
diabetes treatment. The Diabetes Educator 24, 2001-2008.
14. Hayashi, T., H. Maruyama, R. Kasai, K. Hattori, S.
Acknowledgments Takasuga, O. Hazeki, K. Yamasaki and T. Tanaka. 2002.
Ellagitannins from Lagerstroemia speciosa as activators of
glucose traansport in fat cells. Planta Medica 68, 173-175.
This work has been supported by Pusan National
15. Holman, R. R. and R. C. Turner. 1991. Oral agents and in-
University Research Grant for two years (2007.3.- sulin in the treatment of NIDDM. pp. 467-469 In Pickup
2009.2.) and Ho Jeong Research Institute. J. and G. Williams (eds.), Text Book of Diabetes.
Blackwell, Oxford.
16. Kim, S. H., S. H. Hyun and S. Y. Choung. 2006. Anti-dia-
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초록：STZ 으로 당뇨를 유발한 실험쥐에 대한 Psidium guajava L.과 Lagerstroemia speciosa L.

잎추출물의항당뇨효과
노상근․김균하․최원철*
(부산대학교생물학과)

구아바(Psidium guajava L.)와바나바(Lagerstroemia speciosa L.)는항당뇨효과를갖는대표적인약용식물로알려져 있다.

이 들 식 물 은 다 양 한 폴 리 페 놀 (polyphenol) 성 분 을 함 유 하 며 당 뇨 병 의 치 료 에 효 과 적 으 로 작 용 한 다 . 따 라 서
본연구는이들추출물을 streptozotocin (STZ)으로당뇨를유발한실험쥐에섭취시켜항당뇨효과를상호비교하였 다. STZ
으로당뇨를유발한실험쥐에구아바및바나바잎추출물을섭취시킨결과당뇨쥐에비해혈당은약 19-32%, 총콜레스테롤(total
cholesterol) 은 약 24-46%, 중 성 지 방 (triglyceride) 은 22-67% 그 리 고 유 리 지 방 산 (free fatty acid) 은 49-71%
감소하였고또한인슐린분비와췌장에존재하는베타세포의회복도증가되었다 . 한편구아바
추출물의섭취가바나바추출물의섭취에비해항당뇨효과가더높았는데그이유는바나바잎에포함된친수성 폴리페놀
(polyphenol) 성 분 이 에 탄 올 추 출 과 정 에 서 대 부 분 추 출 되 지 않 았 고 또 한 추 출 된 코 로 소 릭 산 (corosolic acid)
은지금까지알려진것과는달리항당뇨활성이낮았기때문이다.