2000 j ethnoph 72

Journal of Ethnopharmacology 72 (2000) 21–27

Hypoglycemic effect of extracts and fractions fromPsacalium 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 Biologicas y de la Salud, Uni6ersidad Autonoma Metropolitana,Unidad Iztapalapa, Apdo. Postal 55-535, 09340 Mexico D.F., Mexico

b Instituto de Quımica, Uni6ersidad Nacional Autonoma de Mexico, Ciudad Uni6ersitaria 04510, Mexico 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 glucosein a dose-dependent manner in normal mice after intraperitoneal administration (PB0.05). This water extract wasmacerated with methanol obtaining a precipitate (WMP fraction), and it was studied in healthy and alloxan-diabeticmice. The WMP fraction showed significant hypoglycemic activity in healthy and mild diabetic mice, but theadministration of this fraction to animals with severe diabetes did not cause any significant decrease in blood glucoselevels. Two polysaccharide components isolated from WMP fraction showed hypoglycemic effect when tested inhealthy mice. © 2000 Elsevier Science Ireland Ltd. All rights reserved.

Keywords: Hypoglycemic plants; Anti-diabetic plants; Medicinal plants; Psacalium decompositum ; Sesquiterpenic compounds

www.elsevier.com/locate/jethpharm

1. Introduction

Diabetes mellitus (DM) is a common disorderassociated with markedly increased morbidity andmortality rate. DM can be defined as a group ofmetabolic diseases characterized by chronic hy-perglycemia resulting from defects in insulin secre-tion, insulin action, or both, resulting in impairedfunction in carbohydrate, lipid and protein

metabolism. Pharmacological treatment of DM isbased on oral hypoglycemic agents and insulin,but these approaches currently used in clinicalpractice either do not succeed in restoring normo-glycemic in most patients or fail after a variableperiod of time (Scheen, 1997; Committee Report,1997).

DM is treated in Mexican traditional medicinethrough anti-diabetic plants (Roman-Ramos etal., 1991, 1992, 1995; Alarcon-Aguilar et al.,1998). One of the most important of these is* Corresponding author.

0378-8741/00/$ - see front matter © 2000 Elsevier Science Ireland Ltd. All rights reserved.

PII: S 0378 -8741 (00 )00202 -6

F.J. Alarcon-Aguilar et al. / Journal of Ethnopharmacology 72 (2000) 21–2722

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 theroots of P. decompositum has shown a decrease onglycemic levels in temporally hyperglycemic rab-bits and has exhibited hypoglycemic effect inhealthy and alloxan-diabetic mice (Alarcon-Aguilar et al., 1997, 2000).

Phytochemical studies have shown thatsesquiterpenic compounds such as cacalol,cacalone, maturin, maturinone, and maturone arethe 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 onhealthy mice (Alarcon-Aguilar et al., 2000).Therefore, it was necessary to direct the attentionto complex mixtures of compounds present inboth organic and aqueous extracts.

We have previously demonstrated the hypo-glycemic effect of one aqueous fraction (WMPfraction) in healthy mice (Alarcon-Aguilar et al.,2000). Therefore, this study had the followingobjectives, (a) to evaluate the hypoglycemic effectof the hexane, methanol, and water extracts of P.decompositum roots in healthy mice; (b) to deter-mine the hypoglycemic effect of a WMP fractionobtained from the water extract of P. decomposi-tum roots in healthy and alloxan-diabetic mice;and (c) to corroborate the hypoglycemic effect oftwo polysaccharide components isolated from theWMP fraction.

2. Material and methods

2.1. Plant material

Roots of P. decompositum were acquired fromthe Sonora Herbal Market at Mexico City. Theidentity was made, with help of a botanist, usingtaxonomic rules and by means of comparisonsamong different herbarium samples of P. decom-positum from MEXU-HERBARIUM (HerbariumIMSSM-Voucher Specimen 11489).

2.2. Preparation of the extracts and compoundsisolated from WMP fraction

The extracts were prepared as previouslydescribed (Alarcon-Aguilar et al., 2000). P.decompositum roots (950.5 g) were ground andextracted four times at room temperature withhexane (3 l, 24 h). The hexane extracts wereconcentrated under reduced pressure andpooled, obtaining 68.18 g (yield 7.17%). Then,the plant residue was extracted four times atroom temperature with methanol (3 l, 24 h).The methanol extracts were concentrated underreduced pressure and pooled, obtaining 86.24 g(yield 9.07%). Finally, the marc was extractedfour times at room temperature with water (3 l,24 h). The water was removed from the extractwith high vacuum obtaining 72.72 g as residue(yield 7.56%). This material was macerated withmethanol (200 ml, 24 h) obtaining a precipitate(WMP fraction, 57 g). The WMP fraction wasanalyzed by high performance thin layerchromatography (HPTLC) and subjected topreparative 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 withPurina nutrition and water ad libitum.Experimental diabetes in mice, subjected toprevious fasting for 18 h, was induced byintraperitoneal administration of alloxan(Rodriguez et al., 1975). The total dose of alloxan(450 mg/kg body weight) was administered inthree injections at intervals of 48 h (150 mg/kgbody weight each time). Seven days after the lastadministration, the animals were fasted for 18 hand blood glucose levels were determined. Theseanimals were included in two experimentalgroups, (a) mild alloxan-diabetic mice, whosebasal glycemia ranged between 200 and 349mg/dl; and (b) severe alloxan-diabetic mice, whosebasal glycemia was equal or higher than 350mg/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 fromP. decompositum roots on fasting-blood glucosele6els in healthy mice

Healthy mice were divided into fifteen groupsof 10–33 animals each (I–XV). Group I and IIserved as controls and received isotonic salinesolution (ISS) or corn oil. Group III received fastaction-insulin (regular insulin) as reference (0.1U.I./kg body weight). The other groups received50, 100, 200, or 400 mg/kg body weight of eachextract. Organic extracts (hexane and methanol)were dissolved in corn oil, and water extract inisotonic saline solution.

2.4.2. Effects of the WMP fraction isolated fromP. decompositum roots on fasting-blood glucosele6els in healthy and alloxan-diabetic mice

Healthy mice were divided into three groups ofeight animals each. Group 1 served as control andreceived ISS; group 2 received tolbutamide (60mg/kg) as reference; and group 3 received 75mg/kg body weight of WMP extract dissolved inISS.

Mild alloxan-diabetic mice were divided intothree groups of six animals each. Group 4 servedas control and received ISS; group 5 receivedtolbutamide as reference (60 mg/kg body weight);the sixth and seventh groups received 150 and 250mg/kg body weight of WMP fraction, respec-tively. Severe alloxan-diabetic mice were dividedinto two groups with six animals each; group 8served as control and received ISS; and finally,group 9 received 250 mg/kg body weight of WMPfraction.

2.4.3. Effects of the main compounds isolatedfrom WMP fraction on fasting-blood glucosele6els in healthy mice

Healthy mice were divided into four groups often animals each. Group A served as control andreceived ISS; group B and C received 200 and 400mg/kg body weight of compound F1, respectively;and group D received 200 mg/kg body weight ofcompound 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 fastinganimals (t=0), and 120 and 240 min after admin-istration substances of test. Glycemia was deter-mined by the glucose-oxidase peroxidase methodwith Haemo-Glukotest 20-800 reagent strips andtheir evaluation was made on a Reflolux-S ligh-meter (Boehringer-Mannheim).

2.5. Statistical analysis

Results were expressed as mean9S.E.M. Thesignificance of the differences between the meansof tests and control studies was established byStudent t-test for independent samples with onetail. P values less than 0.05 were consideredsignificant.

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 notshow significant hypoglycemic effect in healthymice (Table 1). The same doses of the waterextract showed significant hypoglycemic effect inhealthy mice (PB0.05). The hypoglycemic effectof water extract was most evident in the case ofanimals given 200 mg/kg body weight (Table 1).At this concentration, and at dose of 800 mg/kgat 240 min, the water extract was as effective asinsulin (0.1 U.I./kg).

The precipitate obtained from the water extractafter treatment with methanol (WMP fraction),caused a significant decrease in glycemia ofhealthy and alloxan-diabetic mice (Tables 2 and3). The WMP fraction (75 mg/kg) showed a sig-nificant decrease in blood glucose levels of healthymice at 240 min (PB0.005). The WMP fraction(250 mg/kg) also caused a pronounced significantdecrease (PB0.005) in blood glucose levels ofmild 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 1Effect of the hexane, methanol, and water extracts obtained from the roots of P. decompositum on blood glucose levels in fastinghealthy micea

Dose (mg/kg)n Blood glucose (mg/dl, mean9S.E.M.)Study

In fasting 120 min 240 min

– 53.991.8Control (SSI) 50.792.633 51.492.7– 52.992.6Control (corn-oil) 56.993.026 54.793.30.1 U.I./kg 57.192.814 38.291.7***Regular insulin 39.892.1***

50 53.192.9 49.092.9 45.193.0Hexane extract 19100 53.492.619 52.392.9 45.193.2

19 200 54.993.0 63.192.5 45.292.819 400 54.392.5 55.693.7 45.692.8

50 50.792.519 50.792.7Methanol extract 46.393.519 100 54.193.2 50.093.8 42.993.4

200 58.492.419 51.293.4 44.792.7400 46.992.4 50.293.619 38.892.9***

50 53.793.410 57.694.4Water extract 44.793.7100 48.693.1 46.993.410 40.393.2*200 51.792.810 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 2Effect of WMP obtained from water extract of P. decompositum roots on blood glucose levels in fasting healthy mice (n=8)

Dose (mg/kg)Study Blood glucose (mg/dl, mean9S.E.M.)


–Control (SSI) 49.591.9 52.493.7 48.695.148.891.3 37.793.3***60 37.795.3*Tolbutamide47.892.0 41.894.0WMPa 32.892.4***75

a Significantly different from its pre-value in fasting: *PB0.05; **PB0.01; ***PB0.005. WMP, methanol insoluble precipitatefrom water extract.

Table 3Effect of WMP fraction obtained from water extract of P. decompositum roots on blood glucose levels in fasting mildalloxan-diabetic mice (n=6)a

Blood glucose (mg/dl, mean9S.E.M.)Dose (mg/dl)Study


272.8912.7 279.896.2 239.8919.4Control (ISS) –283.2913.3 276.6913.360 125.0924.5***Tolbutamide

150WMP 281.2920.4 261.7923.3 209.8925.7*249.7928.5250 225.8924.9 109.8919.4***



Table 4Effect of WMP fraction obtained from water extract of P. decompositum roots on blood glucose levels in fasting severealloxan-diabetic mice (n=6)a

Dose (mg/dl)Study Blood glucose (mg/dl, mean9S.E.M.)


Control (ISS) – 487.3911.6 473.0925.3 467.2928.7WMP 250 486.5935.9 485.2935.3 458.7948.9

a Significantly different from its pre-value in fasting: *PB0.05.

Table 5Effect of the main polysaccharide fractions isolated from the active WMP fraction of P. decompositum roots on blood glucose levelsin fasting healthy mice (n=10)a

Dose (mg/kg)Study Blood glucose (mg/dl, mean9S.E.M.)


105.094.3Control (SSI) 92.196.1– 94.594.9200F1 105.597.2 89.095.4* 72.492.2***

101.695.6 84.593.3**400 65.892.9***101.094.7F3 82.694.7**200 57.893.4***


The two WMP polysaccharides components iso-lated by pTLC (F1 and F3 fractions) showedhypoglycemic effect in healthy mice. Best resultswere obtained with F3 when administered at 200mg/kg (Table 5).

4. Discussion

The results of this investigation show that thewater extracts of P. decompositum roots exhibits thehighest hypoglycemic effect in healthy mice com-pared with hexane and methanol extracts. In theseanimals, the hypoglycemic effect caused by insulinwarrants the valid of the experimental model.Water extract showed a dose-dependent effectwhen it was administered at dose of 50, 100, and200 mg/kg body weig ht. Doses of 400 and 800mg/kg caused a significant decrease of the glycemia;however, in these cases dose-dependence of theeffect was no apparent. It is likely that the biggerdoses cause some toxic effect, by the presence in thewater extract of some other no hypoglycemicsubstance, which hide the increase of the hypo-

glycemic effect. This result confirms the previouslyobserved hypoglycemic activity of P. decompositumroot 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 alloxandiabetic 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. decompositumroots. They are known to exhibit anti-microbialand allelo-chemical properties (Lotina et al., 1991;Jimenez-Estrada et al., 1992). However, when theanti-diabetic properties of these constituents havebeen evaluated, the results have been contradictory.Cacalol and related compounds did not showhypoglycemic effect on healthy mice (Alarcon-Aguilar et al., 2000), but showed hypoglycemicactivity in genetically altered obese diabetic mice(designed C57BL/61-ob/ob) (Inman et al., 1998).

In the present study, the hexane extract, as wellas the methanol extract, did not show hypoglycemic


effect. These results confirm the results previouslyreported in healthy mice for the sesquiterpenoidscompounds, because they are the most abundantcomponents in the hexane extract (Jimenez-Estrada et al., 1992).

WMP fraction produced an important hypo-glycemic effect in healthy and in mild diabeticmice. In mild diabetic mice tolbutamide was usedas positive control, because in this case a positiveeffect with this hypoglycemic agent implies thepresence of functioning beta cells. It is likely thatWMP fraction also requires the presence of func-tioning b cells, because it was ineffective in severediabetic 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 offour main polysaccharide components. Two ofthem (F1 and F3) showed hypoglycemic effect inhealthy mice (Alarcon-Aguilar et al., 2000). In thepresent research, the hypoglycemic effect of thesetwo components was corroborated in the sameexperimental model.

These last results suggest some additive activitybetween two compounds. There are some caseswhere the pharmacological activity reported in aplant can be explained just by the existence of agroup 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 itsrelative activities (Capasso-Francesco, 1985).Studies will be carried out to evaluate synergicactions between two compounds.

It is interesting to note that many plantpolysaccharides 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 isolatedfrom 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), Trichosantheskirilowii (Ling-Hua and Pei-Gen, 1993), etc. Themajority of these substances have shown hypo-glycemic activity in normal mice and alloxan-in-duced hyperglycemic mice.

In conclusion, the water extract obtained fromP. decompositum roots exhibit hypoglycemic activ-ity in normoglycemic mice. The hexane andmethanol extracts did not show hypoglycemic ef-fect. The WMP fraction isolated from the waterextract exhibited hypoglycemic activity in mildand severe alloxan-diabetic mice. Chemical andpharmacological investigations should be carriedout to evaluate the hypoglycemic activity in dia-betic animals of the main polysaccharide compo-nents isolated from the active WMP fraction.

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