2000 j ethnoph 72

Journal of Ethnopharmacology 72 (2000) 21–27
www.elsevier.com:locate:jethpharm
Hypoglycemic effect of extracts and fractions from
Psacalium decompositum in healthy and alloxan-diabetic
mice
F.J. Alarcon-Aguilar a,*, M. Jimenez-Estrada b, R. Reyes-Chilpa b,
R. Roman-Ramos a
a Departmento de Ciencias de la Salud, Di6ision Ciencias Biolo´gicas y de la Salud, Uni6ersidad Auto´noma Metropolitana,
Unidad Iztapalapa, Apdo. Postal 55 -535, 09340 Me´xico D.F., Mexico
b Instituto de Quı´mica, Uni6ersidad Nacional Auto´noma de Me´xico, Ciudad Uni6ersitaria 04510, Me´xico D.F., Mexico
Received 29 September 1999; accepted 8 February 2000
Abstract
The hypoglycemic effect of the hexane, methanol and water extracts obtained from roots of Psacalium decomposi-tum
(Asteraceae) was investigated in fasting healthy mice. Only the water extract significantly reduced blood glucose
in a dose-dependent manner in normal mice after intraperitoneal administration (PB0.05). This water extract was
macerated with methanol obtaining a precipitate (WMP fraction), and it was studied in healthy and alloxan-diabetic
mice. The WMP fraction showed significant hypoglycemic activity in healthy and mild diabetic mice, but the
administration of this fraction to animals with severe diabetes did not cause any significant decrease in blood glucose
levels. Two polysaccharide components isolated from WMP fraction showed hypoglycemic effect when tested in
healthy mice. © 2000 Elsevier Science Ireland Ltd. All rights reserved.
Keywords: Hypoglycemic plants; Anti-diabetic plants; Medicinal plants; Psacalium decompositum; Sesquiterpenic compounds
1. Introduction
Diabetes mellitus (DM) is a common disorder
associated with markedly increased morbidity and
mortality rate. DM can be defined as a group of
metabolic diseases characterized by chronic hy-perglycemia
resulting from defects in insulin secre-tion,
insulin action, or both, resulting in impaired
function in carbohydrate, lipid and protein
metabolism. Pharmacological treatment of DM is
based on oral hypoglycemic agents and insulin,
but these approaches currently used in clinical
practice either do not succeed in restoring normo-glycemic
in most patients or fail after a variable
period of time (Scheen, 1997; Committee Report,
1997).
DM is treated in Mexican traditional medicine
through anti-diabetic plants (Roman-Ramos et
al., 1991, 1992, 1995; Alarcon-Aguilar et al.,
* Corresponding author. 1998). One of the most important of these is
0378-8741:00:$ - see front matter © 2000 Elsevier Science Ireland Ltd. All rights reserved.
PII: S0378-8741(00)00202-6

22 F.J. Alarcon-Aguilar et al. : Journal of Ethnopharmacology 72 (2000) 21–27
Psacalium decompositum (Gray) Rob. Et Brett.
(Syn. Cacalia decomposita A. Gray), Asteraceae,
popularly known as ‘Matarique’ (Bye, 1986;
Aguilar et al., 1994). Decoction prepared from the
roots of P. decompositum has shown a decrease on
glycemic levels in temporally hyperglycemic rab-bits
and has exhibited hypoglycemic effect in
healthy and alloxan-diabetic mice (Alarcon-
Aguilar et al., 1997, 2000).
Phytochemical studies have shown that
sesquiterpenic compounds such as cacalol,
cacalone, maturin, maturinone, and maturone are
the main constituents of the hexane extract of P.
decompositum roots (Romo and Joseph-Nathan,
1964; Correa and Romo, 1966; Yuste et al., 1976;
Romo de Vivar, 1985). However, these con-stituents
did not show hypoglycemic effect on
healthy mice (Alarcon-Aguilar et al., 2000).
Therefore, it was necessary to direct the attention
to complex mixtures of compounds present in
both organic and aqueous extracts.
We have previously demonstrated the hypo-glycemic
effect of one aqueous fraction (WMP
fraction) in healthy mice (Alarcon-Aguilar et al.,
2000). Therefore, this study had the following
objectives, (a) to evaluate the hypoglycemic effect
of the hexane, methanol, and water extracts of P.
decompositum roots in healthy mice; (b) to deter-mine
the hypoglycemic effect of a WMP fraction
obtained from the water extract of P. decomposi-tum
roots in healthy and alloxan-diabetic mice;
and (c) to corroborate the hypoglycemic effect of
two polysaccharide components isolated from the
WMP fraction.
2. Material and methods
2.1. Plant material
Roots of P. decompositum were acquired from
the Sonora Herbal Market at Mexico City. The
identity was made, with help of a botanist, using
taxonomic rules and by means of comparisons
among different herbarium samples of P. decom-positum
from MEXU-HERBARIUM (Herbarium
IMSSM-Voucher Specimen 11489).
2.2. Preparation of the extracts and compounds
isolated from WMP fraction
The extracts were prepared as previously
described (Alarcon-Aguilar et al., 2000). P.
decompositum roots (950.5 g) were ground and
extracted four times at room temperature with
hexane (3 l, 24 h). The hexane extracts were
concentrated under reduced pressure and
pooled, obtaining 68.18 g (yield 7.17%). Then,
the plant residue was extracted four times at
room temperature with methanol (3 l, 24 h).
The methanol extracts were concentrated under
reduced pressure and pooled, obtaining 86.24 g
(yield 9.07%). Finally, the marc was extracted
four times at room temperature with water (3 l,
24 h). The water was removed from the extract
with high vacuum obtaining 72.72 g as residue
(yield 7.56%). This material was macerated with
methanol (200 ml, 24 h) obtaining a precipitate
(WMP fraction, 57 g). The WMP fraction was
analyzed by high performance thin layer
chromatography (HPTLC) and subjected to
preparative TLC (pTLC) as previously reported,
yielding four main polysaccharides components,
F1, F2, F3, and F4 (Alarcon-Aguilar et al.,
2000).
2.3. Experimental animals
The experimental animals were male adult mice
(CD1-strain) weighing from 25 to 35 g, fed with
Purina nutrition and water ad libitum.
Experimental diabetes in mice, subjected to
previous fasting for 18 h, was induced by
intraperitoneal administration of alloxan
(Rodriguez et al., 1975). The total dose of alloxan
(450 mg:kg body weight) was administered in
three injections at intervals of 48 h (150 mg:kg
body weight each time). Seven days after the last
administration, the animals were fasted for 18 h
and blood glucose levels were determined. These
animals were included in two experimental
groups, (a) mild alloxan-diabetic mice, whose
basal glycemia ranged between 200 and 349
mg:dl; and (b) severe alloxan-diabetic mice, whose
basal glycemia was equal or higher than 350
mg:dl.

F.J. Alarcon-Aguilar et al. : Journal of Ethnopharmacology 72 (2000) 21–27 23
2.4. Biological assays
2.4.1. Hypoglycemic acti6ity of the extracts from
P. decompositum roots on fasting-blood glucose
le6els in healthy mice
Healthy mice were divided into fifteen groups
of 10–33 animals each (I–XV). Group I and II
served as controls and received isotonic saline
solution (ISS) or corn oil. Group III received fast
action-insulin (regular insulin) as reference (0.1
U.I.:kg body weight). The other groups received
50, 100, 200, or 400 mg:kg body weight of each
extract. Organic extracts (hexane and methanol)
were dissolved in corn oil, and water extract in
isotonic saline solution.
2.4.2. Effects of the WMP fraction isolated from
P. decompositum roots on fasting-blood glucose
le6els in healthy and alloxan-diabetic mice
Healthy mice were divided into three groups of
eight animals each. Group 1 served as control and
received ISS; group 2 received tolbutamide (60
mg:kg) as reference; and group 3 received 75
mg:kg body weight of WMP extract dissolved in
ISS.
Mild alloxan-diabetic mice were divided into
three groups of six animals each. Group 4 served
as control and received ISS; group 5 received
tolbutamide as reference (60 mg:kg body weight);
the sixth and seventh groups received 150 and 250
mg:kg body weight of WMP fraction, respec-tively.
Severe alloxan-diabetic mice were divided
into two groups with six animals each; group 8
served as control and received ISS; and finally,
group 9 received 250 mg:kg body weight of WMP
fraction.
2.4.3. Effects of the main compounds isolated
from WMP fraction on fasting-blood glucose
le6els in healthy mice
Healthy mice were divided into four groups of
ten animals each. Group A served as control and
received ISS; group B and C received 200 and 400
mg:kg body weight of compound F1, respectively;
and group D received 200 mg:kg body weight of
compound F3.
In all cases the control substances, extracts,
WMP fraction, and compounds were injected in-traperitoneally
(4 ml:kg body weight). Blood sam-ples
were obtained from the tail vein in fasting
animals (t0), and 120 and 240 min after admin-istration
substances of test. Glycemia was deter-mined
by the glucose-oxidase peroxidase method
with Haemo-Glukotest 20-800 reagent strips and
their evaluation was made on a Reflolux-S ligh-meter
(Boehringer-Mannheim).
2.5. Statistical analysis
Results were expressed as mean9S.E.M. The
significance of the differences between the means
of tests and control studies was established by
Student t-test for independent samples with one
tail. P values less than 0.05 were considered
significant.
3. Results
The hexane and methanol extracts from P.
decompositum root, administered at dose of 50,
100, 200, and 400 mg:kg body weight, did not
show significant hypoglycemic effect in healthy
mice (Table 1). The same doses of the water
extract showed significant hypoglycemic effect in
healthy mice (PB0.05). The hypoglycemic effect
of water extract was most evident in the case of
animals given 200 mg:kg body weight (Table 1).
At this concentration, and at dose of 800 mg:kg
at 240 min, the water extract was as effective as
insulin (0.1 U.I.:kg).
The precipitate obtained from the water extract
after treatment with methanol (WMP fraction),
caused a significant decrease in glycemia of
healthy and alloxan-diabetic mice (Tables 2 and
3). The WMP fraction (75 mg:kg) showed a sig-nificant
decrease in blood glucose levels of healthy
mice at 240 min (PB0.005). The WMP fraction
(250 mg:kg) also caused a pronounced significant
decrease (PB0.005) in blood glucose levels of
mild diabetic mice 240 min after administration.
This effect was as high as the one of tolbutamide
(Table 3). However, in animals with severe dia-betes
the WMP did not show hypoglycemic activ-ity
(Table 4).

Table 1
Effect of the hexane, methanol, and water extracts obtained from the roots of P. decompositum on blood glucose levels in fasting
healthy micea
Study n Dose (mg:kg) Blood glucose (mg:dl, mean9S.E.M.)
In fasting 120 min 240 min
Control (SSI) 33 – 53.991.8 50.792.6 51.492.7
Control (corn-oil) 26 – 52.992.6 56.993.0 54.793.3
Regular insulin 14 0.1 U.I.:kg 57.192.8 38.291.7*** 39.892.1***
Hexane extract 19 50 53.192.9 49.092.9 45.193.0
19 100 53.492.6 52.392.9 45.193.2
19 200 54.993.0 63.192.5 45.292.8
19 400 54.392.5 55.693.7 45.692.8
Methanol extract 19 50 50.792.5 50.792.7 46.393.5
19 100 54.193.2 50.093.8 42.993.4
19 200 58.492.4 51.293.4 44.792.7
19 400 46.992.4 50.293.6 38.892.9***
Water extract 10 50 53.793.4 57.694.4 44.793.7
10 100 48.693.1 46.993.4 40.393.2*
10 200 51.792.8 35.793.3*** 32.993.4***
10 400 51.193.7 38.892.3** 40.492.1*
10 800 59.193.6 45.092.8 22.992.7***
a Significantly different from its pre-value in fasting: *PB0.05; **PB0.01; ***PB0.005.
Table 2
Effect of WMP obtained from water extract of P. decompositum roots on blood glucose levels in fasting healthy mice (n8)
Study Dose (mg:kg) Blood glucose (mg:dl, mean9S.E.M.)
Control (SSI) – 49.591.9 52.493.7 48.695.1
Tolbutamide 60 48.891.3 37.793.3*** 37.795.3*
WMPa 75 47.892.0 41.894.0 32.892.4***
a Significantly different from its pre-value in fasting: *PB0.05; **PB0.01; ***PB0.005. WMP, methanol insoluble precipitate
from water extract.
Table 3
Effect of WMP fraction obtained from water extract of P. decompositum roots on blood glucose levels in fasting mild
alloxan-diabetic mice (n6)a
Study Dose (mg:dl) Blood glucose (mg:dl, mean9S.E.M.)
Control (ISS) – 272.8912.7 279.896.2 239.8919.4
Tolbutamide 60 283.2913.3 276.6913.3 125.0924.5***
WMP 150 281.2920.4 261.7923.3 209.8925.7*
250 249.7928.5 225.8924.9 109.8919.4***

Table 4
Effect of WMP fraction obtained from water extract of P. decompositum roots on blood glucose levels in fasting severe
alloxan-diabetic mice (n6)a
Study Dose (mg:dl) Blood glucose (mg:dl, mean9S.E.M.)
Control (ISS) – 487.3911.6 473.0925.3 467.2928.7
WMP 250 486.5935.9 485.2935.3 458.7948.9
a Significantly different from its pre-value in fasting: *PB0.05.
Table 5
Effect of the main polysaccharide fractions isolated from the active WMP fraction of P. decompositum roots on blood glucose levels
in fasting healthy mice (n10)a
Study Dose (mg:kg) Blood glucose (mg:dl, mean9S.E.M.)
Control (SSI) – 105.094.3 92.196.1 94.594.9
F1 200 105.597.2 89.095.4* 72.492.2***
400 101.695.6 84.593.3** 65.892.9***
F3 200 101.094.7 82.694.7** 57.893.4***
The two WMP polysaccharides components iso-lated
by pTLC (F1 and F3 fractions) showed
hypoglycemic effect in healthy mice. Best results
were obtained with F3 when administered at 200
mg:kg (Table 5).
4. Discussion
The results of this investigation show that the
water extracts of P. decompositum roots exhibits the
highest hypoglycemic effect in healthy mice com-pared
with hexane and methanol extracts. In these
animals, the hypoglycemic effect caused by insulin
warrants the valid of the experimental model.
Water extract showed a dose-dependent effect
when it was administered at dose of 50, 100, and
200 mg:kg body weig ht. Doses of 400 and 800
mg:kg caused a significant decrease of the glycemia;
however, in these cases dose-dependence of the
effect was no apparent. It is likely that the bigger
doses cause some toxic effect, by the presence in the
water extract of some other no hypoglycemic
substance, which hide the increase of the hypo-glycemic
effect. This result confirms the previously
observed hypoglycemic activity of P. decompositum
root water decoction in healthy and alloxan-dia-betic
mice (Alarcon-Aguilar et al., 1997, 2000). P.
decompositum root water decoction caused hypo-glycemic
effect in healthy and in mild alloxan-dia-betic
mice but had a minor effect in severe alloxan
diabetic mice. These data suggest that the P.
decompositum active substances require the pres-ence
of functioning b cells.
Sesquiterpenoids are the major components ob-tained
from the hexane extract of P. decompositum
roots. They are known to exhibit anti-microbial
and allelo-chemical properties (Lotina et al., 1991;
Jimenez-Estrada et al., 1992). However, when the
anti-diabetic properties of these constituents have
been evaluated, the results have been contradictory.
Cacalol and related compounds did not show
hypoglycemic effect on healthy mice (Alarcon-
Aguilar et al., 2000), but showed hypoglycemic
activity in genetically altered obese diabetic mice
(designed C57BL:61-ob:ob) (Inman et al., 1998).
In the present study, the hexane extract, as well
as the methanol extract, did not show hypoglycemic

effect. These results confirm the results previously
reported in healthy mice for the sesquiterpenoids
compounds, because they are the most abundant
components in the hexane extract (Jimenez-
Estrada et al., 1992).
WMP fraction produced an important hypo-glycemic
effect in healthy and in mild diabetic
mice. In mild diabetic mice tolbutamide was used
as positive control, because in this case a positive
effect with this hypoglycemic agent implies the
presence of functioning beta cells. It is likely that
WMP fraction also requires the presence of func-tioning
b cells, because it was ineffective in severe
diabetic mice.
WMP fraction was subjected to thin layer chro-matography
(Alarcon-Aguilar et al., 2000), yield-ing
four main components F1, F2, F3, and F4.
The HPTLC analysis suggested the presence of
four main polysaccharide components. Two of
them (F1 and F3) showed hypoglycemic effect in
healthy mice (Alarcon-Aguilar et al., 2000). In the
present research, the hypoglycemic effect of these
two components was corroborated in the same
experimental model.
These last results suggest some additive activity
between two compounds. There are some cases
where the pharmacological activity reported in a
plant can be explained just by the existence of a
group complex of principles with additive effects,
isolated from the same whose actions are particu-larly
difficult to reproduce with pure substances.
In these cases, the principles are different in its
relative activities (Capasso-Francesco, 1985).
Studies will be carried out to evaluate synergic
actions between two compounds.
It is interesting to note that many plant
polysaccharides have been reported to exhibit hy-poglycemic
effects (Ling-Hua and Pei-Gen, 1993;
Marles and Farnsworth, 1995; Perez et al., 1998).
Some hypoglycemic polysaccharides were isolated
from the roots of Panax ginseng (Oshima et al.,
1985), Lithospermum erythrorhizon (Konno et al.,
1985), Dioscorea japonica (Hikino et al., 1986a),
Oryza sati6a (Hikino et al., 1986b), Trichosanthes
kirilowii (Ling-Hua and Pei-Gen, 1993), etc. The
majority of these substances have shown hypo-glycemic
activity in normal mice and alloxan-in-duced
hyperglycemic mice.
In conclusion, the water extract obtained from
P. decompositum roots exhibit hypoglycemic activ-ity
in normoglycemic mice. The hexane and
methanol extracts did not show hypoglycemic ef-fect.
The WMP fraction isolated from the water
extract exhibited hypoglycemic activity in mild
and severe alloxan-diabetic mice. Chemical and
pharmacological investigations should be carried
out to evaluate the hypoglycemic activity in dia-betic
animals of the main polysaccharide compo-nents
isolated from the active WMP fraction.
References
Aguilar A., Camacho J.R., Chino S., Jacques P., Lopez M.E.,
1994. Herbario medicinal del Instituto Mexicano del Se-guro
Social (IMSS). Mexico, p. 55.
Alarcon-Aguilar, F.J., Roman-Ramos, R., Jimenez-Estrada,
M., Reyes-Chilpa, R., Gonzalez-Paredes, B., Flores-Saenz,
J.L., 1997. Effects of three Mexican medicinal plants (As-teraceae)
on blood glucose levels in healthy mice and
rabbits. Journal of Ethnopharmacology 55, 171–177.
Alarcon-Aguilar, F.J., Roman-Ramos, R., Perez-Gutierrez,
M.S., Aguilar-Contreras, A., Contreras-Weber, C.C., Flo-res-
Saenz, J.L., 1998. Study of the anti-hyperglycemic ef-fect
of medicinal plants used as anti-diabetics. Journal of
Ethnopharmacology 61, 101–110.
Alarcon-Aguilar, F.J., Jimenez-Estrada, M., Reyes-Chilpa, R.,
Gonzalez-Paredes, B., Contreras-Weber, C.C., Roman-
Ramos, R., 2000. Hypoglycemic activity of root water
decoction, sesquiterpenoids and one polysaccharide frac-tion
from Psacalium decompositum in mice. Journal of
Bye, R.A. Jr, 1986. Medicinal plants of the Sierra Madre:
comparative study of Tarahumara and Mexican market
plants. Economic Botany 40, 103–124.
Capasso-Francesco, 1985. Medicinal plants: an approach to
the study of naturally occurring drugs. Journal of
Committee Report, 1997. Report of the Expert Committee on
the diagnosis and classification of diabetes mellitus. Dia-betes
Care 20, 1183–1197.
Correa, J., Romo, J., 1966. The constituents of Cacalia decom-posita
A. Gray. Structure of maturin, maturinin, maturon,
and maturinone. Tetrahedron 22, 685–691.
Hikino, H., Konno, C., Takahashi, M., Murakami, M., Kato,
Y., Karikura, M., Hayashi, T., 1986a. Isolation and hypo-glycemic
activity of dioscorans A, B, C, D, E, and F;
glycans of Dioscorea japonica rhizophores. Planta Medica
52, 168–171.
Hikino, H., Murakami, M., Oshima, Y., Konno, C., 1986b.
Isolation and hypoglycemic activity of oryzarans A, B, C,
and D; glycans of Oryza sati6a roots. Planta Medica 52,
490–492.

Inman, W.D., King, S.R., Evans, J.L., and Luo, J., 1998.
Furanoeremophilane and eremophilanolide sesquiterpens
for treatment of diabetes. US Patent Number 5,747,527.
Jimenez-Estrada, M., Cruz, R., Valdez, J., Leon, J., Alar-con,
G., Svestarova, B., 1992. Actividad antimicrobiana
del cacalol y sus derivados. Revista Latinoamericana de
Quı´mica 22, 14–17.
Konno, C., Mizuno, T., Hikino, H., 1985. Isolation and
hypoglycemic activity of lithospermans A, B, and C, gly-cans
of Lithospermum erythrorhizon roots. Planta Medica
51, 168–171.
Ling-Hua, Z., Pei-Gen, X., 1993. Recent advances in studies
of antihyperlipaemic and antihyperglycaemic compounds
from Chinese traditional and herbal medicines. Phy-totherapy
Research 7, 217–226.
Lotina, B., Roque, J.L., Jimenez-Estrada, M., Aguilar, M.,
1991. Inhibition on oxygen evolution by cacalol and its
derivatives. Zeitschrift fur Naturforschung 46, 777–780.
Marles, R.J., Farnsworth, N.R., 1995. Antidiabetic plants
and their active constituents. Phytomedicine 2, 137–189.
Oshima, Y., Konno, C., Hikino, H., 1985. Isolation and
hypoglycemic activity of panaxans I, J, K and L; glycans
of Panax ginseng roots. Planta Medica 52, 168–171.
Perez, G.R.M., Zavala, S.M.A., Perez, G.S., Perez, G.C.,
1998. Antidiabetic effect of compounds isolated from
plants. Phytomedicine 5, 55–75.
Rodriguez, H., Perez, R.M., Mun˜ oz, H., Perez, C., Miranda,
R., 1975. Induccio´n de diabetes en raton por medio de
aloxana. Acta Medica XI, 33–36.
Roman-Ramos, R., Flores-Saenz, J.L., Partida-Hernandez,
G., Lara-Lemus, A., Alarcon-Aguilar, F., 1991. experi-mental
study of the hypoglycemic effect of some antidia-betic
plants. Archivos de Investigacio´n Medica 22,
87–93.
Roman-Ramos, R., Alarcon-Aguilar, F., Lara-Lemus, A.,
Flores-Saenz, J.L., 1992. Hypoglycemic effect of plants
used in Mexico as antidiabetics. Archives of Medical Re-search
23, 59–64.
Roman-Ramos, R., Flores-Saenz, J.L., Alarcon-Aguilar, F.,
1995. Anti-hyperglycemic effect of some edible plants.
Journal of Ethnopharmacology 48, 25–32.
Romo de Vivar, A., 1985. Productos naturales de la flora
mexicana. Limusa, Mexico, p. 69.
Romo, J., Joseph-Nathan, P., 1964. The constituents of Ca-calia
decomposita A. Gray. Structures of cacalol and
cacalone. Tetrahedron 20, 2331–2337.
Scheen, J.A., 1997. Drug treatment of non-insulin-dependent
diabetes mellitus in the 1990s. Achievements and future
developments. Drugs 54, 355–368.
Yuste, F., Diaz, E., Walls, F., 1976. The structure of
cacalone. Journal of Organic Chemistry 41, 4103–4106.
.

2000 j ethnoph 72

Recommended

Recommended

More Related Content

What's hot

What's hot (20)

Similar to 2000 j ethnoph 72

Similar to 2000 j ethnoph 72 (20)

More from Ricardo Reyes Chilpa. Instituto de Química. UNAM

More from Ricardo Reyes Chilpa. Instituto de Química. UNAM (20)

Recently uploaded

Recently uploaded (20)

2000 j ethnoph 72