Indo. J. Phar. Scie. Tech. Vol. 2, No. 1, 8-12 (2015). http://dx.doi.org/10.15416/ijpst.v2i1.7805
Nurmeilis1, Dyah A. Woro1, Atiek Soemiati1, Anis Khoirunisa2
1Faculty of Medicine and Health Science, UIN Syarif Hidayatullah, Bandung, West Java, Indonesia
2Faculty of Pharmacy, Universitas Padjadjaran, Sumedang, Jawa Barat, Indonesia
Korespondensi : email@example.com (Nurmeilis)
Kata Kunci: E. longifolia, diuretik, interaksi herbal-obat, irbesartan, toksisitas akut
The hypertension prevalence based on measurement and history of disease was 32.2% with risk factors are geriatric, man, and obesity.1 There are many conventional antihypertensive which have side effects.2 The European Society of Cardiology/ Hypertension (ESC/ESH) and JNC (Joint National Committee) VII recommend the diuretic as one of antihypertensive drugs.3
The use of herbal as supplement and alternative treatment has been increasing in recent years. People combined herbal and drugs without any consultations with the doctor previously. Many people have the mistaken that being all natural herbs are safe but it is not so. There are many herbs may interact with conventional medicine normally taken simultaneously that obtain serious side reaction. However, there are many herbs have synergist effect for particular drugs.4
Eurycoma longifolia Jack have been known by the local names Tongkat Ali in Malaysia and Pasakbumi in Indonesia. It is been used as aphrodisiac, antimalaria, and antidiabetic empirically. Blood pressure can be decreased by intravena injection of E. longifolia aqueous extract.5 The toxicity of E. longifolia which is described by LD50 value is higher than 5000 mg/kg, there is no significant pathologic on kidneys, liver and testes of rat.6 E. longifolia has many bioactive compounds include the alkaloids and quassinoids form in a major portion (such as eurycomaoside, eurycolactone, eurycomalactone, pasakbumin-B, and eurycomanone).7
Both of the effectiveness herbal-drug combination with modern pharmaceuticals and the possible adverse effects from herb– drug interactions remain to be verified. This study was carried out to investigate the diuretic effect of E. longifolia extract and its combination with irbesartan in normal rats, also to determinate the LD50 value and the toxic effects that influence the organ weight and histopathology.
The materials are Male SD rats (175 200 g) and male ddY mice (20 40 g) were purchased from animal house of Bogor Agricultural Institute. They were maintained under standard conditions of temperature and humidity. E. longifolia powder was obtained from PT. Deltomed, Solo and Irbesartan was purchased from PT. Indofarma, Bekasi.
E. longifolia powder as much as 800 g was extracted by maceration method then stirring occasionally. Maceration method used aquadest solvent that distilled until a limpid maserat. Aqueous extract (maserat) was concentrated by evaporating solvent using a rotary evaporator at temperature 40 oC to obtain a thick extract, dried with oven vaccum. Five senses used to describe the organoleptic characteristics of extract observed such as the shape, color, smell, and taste.
The assessment method for diuretic activity based on Sreelakshmi et al.8 Six groups of six mice each group were fasting and not given water for eighteen hours before the experiment. Rats were placed in metabolic cages for 24 hours, without food or water during this period. The treatment group was given 40 mg/kgBW E. longifolia aqueous extract orally, 40 mg/kgBW irbesartan dose and the combination of E. longifolia aqueous extract and irbesartan (20, 40 and 80 mg/kgBW). Urine volume is recorded at 2, 4, 6 and 24 hours post administration. Sodium and potassium ion concentrations in urine samples were determined by AAS (Atomic Absorbance Spectrometer).
Male ddY mice were divided into seven groups of six animals each. Control group received a normal saline (2 mL/kg) and the other groups received 1, 2, 4 and 8 g/kg combination of E. longifolia extract and irbesartan, respectively. Immediately after dosing, the animals were observed for their behavior continuously four hours for the first. They were kept under observation up to 14 days after extract administration to find out the mortality, signs toxicity and body weight.
The result of sodium and potassium ion concentrations in urine samples shows that all of groups test increased volume of urine excreted in 24 hours. The concentration of Na+ and K+ ion increase significant (p<0.05) from control normal group (Table 1).
The combination dose of E. longifolia water extract and irbesartan (1, 2, 4 and 8 g/kgBW) caused the mortality of two mice at the highest dose (8 g/kgBW) with 40% mortality response during the 14 hours observation period (Table 2). The mice showed no change in behavior and clinical signs of toxicity such as passive, ataxia, or tremor. LD50 values were determined by probit analysis and regression equation was Y=a+bx. Y=-1088+1.39x, where x is the antilog dose (LD50)= 23.981 g.
The results of histopatology showed in Table 3. The histology result of liver and heart of acute toxicity looked normal, but there is atrophy of the glomeruli in the kidneys of two mice (8000 mg/kg treatment groups).
Diuretics are drugs that capable of increasing levels of urine as well as the electrolyte output, so they are useful in the treatment of diseases that related with the retention of fluids like high blood pressure, or heart failure, and nephrotic syndrome. There are correlations between the volume of urine and the concentration of Na+, this aspect is logical because the mechanism of action of diuretic drugs is to decrease the tubular reabsorption of this ion, it produces the dragging of the osmotic equivalent of water, other explanation that can support this, is the high ion concentrations in this medicinal plants.
There is no significant difference between single dose of E. longifolia extract (dose: 40 mg/kgBW) and irbesartan (dose: 40 mg/kgBW) but the combination of E. longifolia extract and irbesartan has lower activity than single extract. So the extract of E. longifolia has the same activity with irbesartan, but if its combination the effect of diuretic decrease. It is caused possible by interaction of drug that influences the metabolism. So, the E. longifolia extract has same activity with irbesartan, but when combined, the diuretic effect decreases. It is probably caused by drug interactions that affect the metabolism. In addition, irbesartan is one of antihypertensive drugs that has a narrow therapeutic index so that if consumed concomitant with other drugs or herbal therapies may alter the effects of irbesartan. For example irbesartan with garlic or ginseng is able to decrease its effect.9
Based on the substance of the table toxicity scale, in practical classification of toxic compounds of the LD50 greater than 15 g/kg,10 so that the combination of E. longifolia extracts and irbesartan is safe.
Based on the result of histopathology, liver and heart looked normal, but there is atrophy of the glomeruli in the kidneys of two mice (8000 mg/kg treatment groups). These degenerative changes are an early manifestation of cell injury. This incident is reversible because it seen improvement in cells. However, if injuries persist, so the necrosis will occur. There is no significant change in pathological effects of all tested dose levels during the observation period of 14 days.
Table 1 Volume of Urine Excreted, Na+ and K+ Concentration (Mean ± Standard Deviation, n=6)
Table 2 Respon of Mortality
Table 3 Results of Histopathology
All of the tested group have significant diuretic activity (p<0.05) in comparison with control normal group. A single dose of E. longifolia extract (40 mg/kgBW) and irbesartan (dose: 40 mg/kgBW) had no significant difference. But the combination of E. longifolia extract and irbesartan are lower activity than single extract.
Based on table toxicity scale, LD50 value of combination E. longifolia extract with irbesartan is 23.981 g/kg and can be categorized as nontoxic compound. The histopathological result showed that there is no significant pathologic effect in the kidneys, liver, and heart.
We thank to the Center of Research and Development of UIN Syarif Hidayatullah Jakarta for providing the grant for this study.
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