DRUGSEXPTL.CLiN. RES. XXVI(3) 89-93 (2000)~~..;.J) J;i;1tViJplt:, ..//

ANTIHYPERTENSIVE AND ANTIHYPERLIPIDEMIC EFFECTSOF ACHILLEA WILHELMSII

ASGARY S., NADERI G.H., SARRAFZADEGAN N., MOHAMMADIFARD N.,MOSTAFAVIS., VAKILI R.

Isfahan Cardiovascular Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.

Summary: Achillea wilhelmsii C. Koch (Asteraceae) is widely found in different parts of Iran. This plant is fullof fIavonoids and sesquiterpene lactones, which have been shown to be effective in lowering blood lipids andhypertension. We conducted a double-blind placebo controlled clinical trial to study the antihyperlipidemicand antihypertensive effects of Achillea drops. We randomly selected 120 men and women, aged 40-60years, and divided them in two distinct groups of moderate hyperlipidemic and primary hypertensive subjects.They were treated either with hydroalcoholic extract or with placebo in the form of 15-20 drops twice daily formore than 6 months. Blood pressure and serum lipids (total cholesterol, triglyceride, low-density lipoprotein(LOL) cholesterol and high-density lipoprotein (HOL) cholesterol) were measured in the groups for 3 periodsof 2 months each. The mean and standard deviation of alternations in these variables between the group tak-ing placebo and thC!!ttaking drugs was calculated by Student's Hest. The results showed a significantd,ecreasein triglycerides after of 2 months while decreases in triglycerides, total cholesterol and LOL-choles-terol were significant after 4 months. Levels of HOL-cholesterol were significantly increased after 6 months'treatment. A significant deprease was observed in diastolic and systolic blood pressure after 2 and 6 months,respectively (p <0.05).

Introduction

Achillea wilhelmsii C. Koch (Asteraceae) is a wildgrowing herb widely distributed in different parts ofIran, especially in the central and western areas (1-3).It has long been used in traditional Persian medicineas a tonic and carminative substance in the treatment

Address fO( correspondence: S. />sgary.lJ;fahanCardiovascularR~ch Center. Isfa/'.an University of Me<f~ Sciences. 'P.O. Box

.81465-1148. Isfahan, Iran. .

Tel.: 98-31.460807/98.31-461826 Fax.: 98-31459023

E-mail: isf.carvascrc@asia-mail.com

0378-6501/2000/300089+5 $02.50/0

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of pulmonary conditions (4). The aerial parts of theplant; such as the leaves, flowers, twigs and fruit areused for therapeutic applications (5-7). Phytochemicalstudies reveal that this species is a rich source offIavonoids and sesquiterpene lactones, which have aremarkable effect on blood lipids (5-10). Some highlydiverse substances were isolated from this plantincluding alkaloids, volatile oils, saponin, tannins,resin, sterols, aJkamids,and carbohydrates (8).

Flavonoids have proved to be strong,: free radicalscavengers that affect .cellular phosphorylation andinhibit lipid peroxidation, both enzymatically andnonenzymatically (11-14). The antioxidant com-

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pounds of flavonoids and sesquiterpene provokechanges in fats and proteins through their own dif-ferent mechanisms (15-16). Sesquiterpene lactonecompounds have been shown to have numerous

biological activities due to their strong electrophilicsystem, such as analgesic, antiinflammatory andantihyperlipidemic properties (3, 5, 6). Also, plantalkaloids decrease arterial blood pressure and haveantispasmodic, enteric and uterine contractioninhibitory and antifebrile properties (17-19).

Regarding the mechanism of the above mentionedactive factors, this genus may reduce cardiovasculardisease and its associated risk factors such as

hypertension, hyperlipidemia and atherosclerosis. Inthe present study we report the antihypertensive andantihyperlipidemic effects of A. wilhelmsii.

Materials and methods

Plant gathering. Plant material was.supplied from awild species growing in Chatrood village in theprovince of Kerman in southeast Iran, at an altitudeof 1,840 m in June 1995. The plant was identified inthe Botany Garden of Fillips Marburg University inGermany and in the Karaj Forestry and PasturageResearch Center in Iran.

Preparation of herbal drops. Air-dried powder fromaerial parts of flowering A. wilhelmsii were extractedwith the 48 h percolation method using 70% ethanolat a ratio of 1:8, respectively (1 g of dried herb to 8ml of 70% ethanol) (20).

Preparation of placebo drop. Alcoholic placebo.free from the plant material, was used to support thebiological activity of the herb species through aplacebo controlled, double-blind clinical trial.

Physicochemical controls. Physicochemical con-trol tests were performed on the extract indicating

the percentage of flavonoids, pH, density. alcoholicdegree, dry left-over, color, smell and taste, asshown in Table I (20-21). The flavonoid extracts were

determined by absorbance at 425 nm as follows:

625 x extract absorbance

A'% x 1 cm x powder weight

w~ere A 1"10= hyperoside absorbance at 500.

Table I Results of the chemical and physical specifications of the

herbal drops

Type of test ResuRs

The dry leftover

Flavonoids

Percentage of alcoholpHDensityColorSmellTaste

3-4 g/100 ml

100-110 mg/100 ml

525.47

0.8981

LightgreenAlmost bright

Bitter

Selection of patients. This study was a double-blindplacebo controlled clinical trial. Subjects were select-ed randomly, based on the record numbers ofpatients referred to the regional Isfahan Carc:Jio-vascular Research Center in Isfahan, Iran. The.patients were 120 equally randomized men andwomen, aged 40-60 years. They were divided intotwo distinct groups, one group of 60 patients withmild hypertension (systolic blood pressure: 140-160mmHg; diastolic blood pressure: 90-95 mmHg) anc.'another group of 60 patients with moderate hyperlipi-demia (total cholesterol: 200-300mgldl; triglyceride:

.200-400 mgldl). Those who w~re undergoing medicaltreatment or dietary therapy were excluded. Exclusion

criteria included uncontrolled hypertension and

uncontrolled, hereditary or secondary hyperlipidemia.

Patients who had both risk factors (hypertension and

hyperlipidemia), coronary artery disease or any other

disorders were also excluded. Blood pressure was

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Asgary S. et al.

Table II Comparison of changes in plasma lipid levels and blood pressures between two drug and placebo groups for hypertensive and

hyperlipidemic subjects in different time intervals

'PEach factor was compared to its initial form (before treatment). "Minus sign indicates decrease of factors compared to initial values.

Concerning the limitations of this study, it would

have t)een better to investigate the effect of Achillea

extract on. patients with severe hypertension andhyperlipidemia. In addition, the time taken to lower

blood pressure is. fairly long. An effect could be

achieved earlier by' differen~' extract' processing,

increasing the recOmmended dosage or by preparing

other phytopharmaceutical forms of this plant extract..Also, there are several reasons for the above men-

tioned differences not being significant: the validity of

our data is based on the belief that our patients used

the herbal drop according to the instruction (self-

reporting). A biomarker is needed to verify the results

in our subjects. Choosing an appropriate sample size,

adjusted to the different clinical characteristics of thepatients, is another important consideration in achiev-

ing reliable results on Achillea medicine.

References

(1) Rechinger K.H. Compositae VI-Anthemideae. Akademische

Druck-U. Verlagsanstalt, Graz-Austria. Rora lranica. 158, 53, 1986.

(2) Ghahraman A. "Chromatophytes of Iran (Plant Systematic).

Vol. 3". Tehran University Press. Tehran, 1995. p. 685.

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Variables Drug group Placebo group p-value

Mean :t SO (n =30) Mean:t SO n =30)

Total cholesterol (mg/dQ'PAfter 2 months "-o.8:t 12.1 2.3:t 11.1 NS

After 4 months -31.7 :t 84.4 2.9:t 41.2 0.04

After 6 months -38.8 :t 59.4 4.6:t 91.4 0.3

LDL-cholesterol (mg/dQAfter 2 months -16:t45.1 2.6:!: 32.3 NS

After 4 months -31.4 :t 32.4 -4.2 :t 39.3 0.001

After 6 months -35.6 :t 42.6 3.4 :t 28.5 <0.0001

HDL-cholesterol (mg/dQAfter 2 months 17 :t 24.3 0.7 :t3.6 NS

After 4 months 17:t 8.1 15.3:!:5.6 NS

After 6 months 20.4:t 9.1 -4.3 :t 8.9 <0.0001

Triglycerides (mg/dl)After 2 months --£ :t 24.3 2.9 :t 6.51 0.05

After 4 months --£7.3 :t 56.4 8.4 :t 64.9 <0.0001

After 6 months ... -75.3 :t 48.2 14.5:t 52.3 <0.0001

Systolic blood pressure (mmHg)After 2 months -5:t 10.2 -2.9 :!:5.0 NS

After 4 months -11.2 :t 9.8 -4.7:t 13.6 NS

After 6 months -14.1 :t 10.9 -3.8 :t 8.6 0.005

Diastolic blood pressure (mmHg)After 2 months -B.3 :t 5.8 -o.6:t 7.8 0.003

After 4 months -16.9:t 5.0 -1.4 :t9.9 0.001

After 6 months -14.7:t 6.2 -2.6 :t 8.6 <0.0001

c

Antihypertensive and antihyperlipidemic effects of Achillea wilhelmsii

(3) Zahedi A. "Plant Dictionary" No. 509. Tehran University

Publications, Tehran, 1958, p. 9.

(4) Zargary A. "Medical plants". Vol. 3. Tehran University

Publications, Tehran, 1992, pp. 106-116.

(5) Zrtterl-Eglseer K., Jurenitsch J. Sesquiterpene lactone of Achillea

setacea with antiphlogestic activity. PlantaMed., 57(5).444, 1991.(6) Kastner U.. Sosa S. Anti-edematous activity of sesquiterpene

lactones from different taxa of Achillea millefolium group. PlantaMed., 59 (Suppl.), 95, 1993.

(7) Ahmed A.A.. EI-Sayed N. Ravonoids of Achillea santolina and

Achillea fragrantissima. Rev.Latinomar.Quim., 20(1),5, 1989.(8) Elman M.A., Hassan A.B., AUa A.H., Fahmy G.E.

Phytochemical and pharmocological studies on Achillea santolina.

Egypt. Y. Vet. Sci., 2(24), 181, 1987.

(9) BrunkeE.J., HammerschmidtEJ., Aboutabl E.A. Volatile con-stituents of Achillea wilhelmsii C. Koch (syn. Achillea santolina auct.

mult.) from Egypt and Turkey.Proc. Inl. Symp. Essent. Oils. Prog.Essent. Oil Res.. 16. 85, 1986.

(10) Gerger H., Hofer O. Highly unsaturated isopentyl amides

from Achillea wilhelmsii. Nat. .'pro., 50(6), 1100, 1987.

(11) Harborne J.B. "The' Flavonoids". Chapman and Hall,

London. 1994:J'>p.638-639.

(12) Huguet AT. ManezS.. AlcarazM.J. Superoxide scavengingproperties of flavonoids in a nonenzymatic system. Z. Naturforsch.,45, 19. 1990.

(13) Costantino L., Albasini A., Rastelli G., Benvenuti S. Activity of

polyphenolic crude extract as scavengers of superoxide radicalsand inhibitors of xanthine oxidase. Planta Med., 58(4),342. 1992.

(14) Geofrey A. Changing strategies in natural products chem-

istry. Phytochemistry. 40(6), 1585, 1995.

(15) Navab M., Fogelman M.. Berliner Y., et al. Pathogenesis of

atherosclerosis. Am. J. Cardiol., vol. no?, 76, 1995.

(16) Eloy-RodriguezG.H.N., MitchellJ.C. Biologicalactivities ofsesquiterpenelactones. Phytochemistry, 15, 1573. 1976.

(17) AI-Hindawi MK. AI-Deen I.H., Nabi MH. Ismail MA Anti-

inflammatory activity of some Iraqi plants using intact rats.

Ethnopharmacol.,26(2). 163. 1989.

(18) Harborne J.B. "Phytochemical Methods. AGuideto ModernTechniques of Plant Analysis".Chapman and Hall.London. 1998.pp. 203-214.

(19) Popovici L., Braha S., Gafiteanu E. Anti-infective agents.

Farmacia,34, 141, 1986.

(20) List P.H. "Technologie Pflanzlieher Arzneizuberelturgen".

Wissenschaftliche Verlagsgesllschaft mbH, Stuttgart, 1982. p.

263. ._' ..' .'-(21) W~chtl M. "Diepharmakognostische Chemische Analyse".

"-Akademische Werla9sgesellschaft, Frankfurt. 1971..p.. ~8.5 .(22) Petkov V., Manolov P. Pharmacological studies on sub-

stances of plant origin with coronary dilating and' anti-arrhythmic

action. Comp. Med. East West., 6(2), 123, 1978.

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Antihypertensive and antihyperlipidemic effects of Achillea wilhelmsii

measured by the World Health Organization's stan-dardized method using. a random zero sphygmo-manometer to determine mildly hypertensive subjects(22). For the hyperlipidemic group total cholesterol,triglycerides, low-density lipoprotein (LDL) cholesteroland high-density lipoprotein (HDL) cholesterol wereinitiallymeasured when patients were in a 14 h fastingstate with enzymatic standardized method by anELAN 2000 autoanalyzer. Blood lipids and bloodpressure measurements were repeated for eachpatient after 2,4 and 6 months of medication.

All patients were adjusted for age, body massindex and occupation and were divided into the drugor placebo groups in two separate antihyperlipidem-ic and antihypertensive studies. They were treatedorally twice daily in a volume of 15-20 drops of theplacebo or the plant extract for up to 6 months.Fifteen to twenty drops of the plant extract equal1-1.1 mg flavonoids on the basis of hyperoside.

Statistical analysis. Data was entered by EPI6soft-ware and statistically analyzed in SPSS/WIN.Student's t-test was used to compare alterations inlipid profiles and systolic blood pressure between thedrug and placebo groups. To compare diastolicblood pressure, due to lack of normal distribution inthe diastolic blood pressure data, the nonparametrictest (Wilcoxon's) was used in the hypertensivegroups. These comparisons wer~ done after 2, 4and 6 months of treatment.

Results

In the present study, some interesting results were'obtained through a 6-month treatment with theIranian species of A. wilhelmsii. Comparison ofresults indicates that in hypertensive patients, dias-tolic blood pressure showed a statistically significantdecrease after 2 months' treatment and that thisdecrease continued until the end of the 6-month

treatment period (p =0.003). However, systolic bloodpressure showed no significant alteration after 2, 4 or6 months' treatment (p =0.005).

In the hyperlipidemic group, a significant decreasein triglyceride levels was observed after 2 months'treatment in the group taking drugs (p <0.05). Whenlevels of LDL- and total cholesterol were assessed,

the plant extract showed its possiqle effects througha significant drop in these levelsafter 4 months (fable

II).However, HDL-cholesterol. levels increased signif-icantly only after 6 months' treatment (p <0.05).These findings could be explained by the therapeuticimprovements in lipid profiles after at least 2 monthsof d.rugadministration. Moreover, an antihypertensivetrend for the plant extract might be expected after 6months of oral administration.

Except for the bitter taste of the extract, noadverse effects, such as constipation, hiccups,

h~adaches, vomiting or nausea, were seen in thepatients. In addition, the plant extract produced ben-eficial effects on alimentary discomforts.

Discussion

Several phytochemical studies have been per-formed on the. therapeutic effects of the Achilleaspecies, mainly for the major components offlavonoids and the sesquiterpene lactone (5-10). AI-Hindawi et al. (17) found that the plant has antihy-pertensive and antiinflammatory effects on rats,probably due to its alkaloids.

Antihypertensive and antihyperlipidemic medica-tion are.essential to diminish the high morbidity af)dmortality due to' cardiovascular.disease. Because of'the body's adaptation to the chemical composition ofplants and the low incidence of adverse effects asso-ciated with plant extract administration comparedwith the adverse effects and high expense of syn-thetic chemical medicine. the use of phytotherapywill probably increase. 9