Effect of alcoholic extract of Lepidium meyenii (Maca) on testicular function in male rats

Gustavo F. Gonzales, Julio Rubio, Arturo Chung, Manuel Gasco, Leon Villegas-

Department of Biologicnl and physiological Sciences. Faculty of Sciences and Philosophy and Instituto de

Investigaciones de la Altura

Department of Biochemistry, Molecular Biology and Pharmacology, Universidad Peruana Cayetano Heredia.

Lima, Peru

Keywords: Lepidium meyenii; transillumination; spermatogenesis; sperm count

Abstract Aim: To evaluate the effect of the alcoholic extract of Lepidium meyenii (Maca) on the spermatogenesis in male rats. Methods: In Holtzman rats, Maca alcoholic extract (5%)was given by oral route at doses of 48 mg/day or 96 mg/day for 7 days, 14 days and 21 days. Testicular function was assessed by measurements of lengths of different stages of seminiferous epithelia and by epididymal sperm count. Results: Ethanolic extract of Maca increased the length of stages IX-XI of seminiferous epithelium at treatment day 7, day 14 and day 21.Progression of spermatogenesis was evident only after day 21 when lengths of stages XII-XIV of seminiferous epithelium were increased; at day 7 and day 14, no important change in spermatogenesis was observed. Epididymal sperm count wasincreased with 48 mg/day at all times. With 96 mg/day an increase in sperm count was observed at day 7, but it was reduced at day 14 and day 21 of treatment. Serum testosterone levels were not affected. Conclusion: The alcoholic extract of Maca activates onset ant progression of spermatogenesis at 48 mg/day or 96 mg/day in rats. (Asian J Androl 2003 Dec; S: 349-352)

1 Introduction

Lepidium rneyenii (Maca) is a perennial plant cultivated in the Andean mountains over 4,000 m altitude.The root traditionally has been used for its fertility-enhancing properties as described in the Spaniard conquestChronicles of Peru in 1653 t1]. Probably Maca was domesticated in San Blas, Junin, 1300 to 2000 years ago[2]. Recently, it has been demonstrated that aqueousextract of Maca improves spermatogenesis and semen parameters [3, 4]. In the present study, the effect of the alcoholic extract of Maca on spermatogenesis and epididymal sperm count was assessed.

2 Material and methods

2.1 Plant material

Lepidium meyenii was obtained from Carhuamayo,Junin at the Central Andes of Peru at 4,000 m altitude and authenticated by Irma Fernandez, Assistant Professor and botanist of the Department of Biochemistry,Molecular Biology and Pharmacology, Universidad Peruana Cayetano Heredia (Peru)

2.2 Extraction

The fat of the dried and powdered roots of Lepidium meyenii (750 g) was removed with soaking in hexane at room temperature (20 ℃) for 3 days. The residue obtamed (14.5 g) was extracted with 1,500 mL absolute ethanol for 72 h at room temperature. The filtrate was

concentrated under reduced pressure at 40 ℃ with Rotovapor (Biichi, Water bath B-480, Switzerland) and the final dry extract weighed 53.1 g. The dry extract,2.4 g and 4.8 g, were dissolved separately in 100 mL of 5%ethanol with resultant solutions of 24 mg/mL (solution

A) and 48 mg/mL (solution B)

2.3 Animals

Three一month-old male Holtzman rats, weighing 295.4 gplusmn;7.8 g (SD), from the animal house of the Instituto de Investigaciones de la Altura, Universidad Peruana Cayetano Heredia were used. Animals were housed under standard conditions (12 h light/12 h dark,22℃). Rats were fed Purina laboratory chow(Agribrands Purina Peru S.A, Lima, Peru) and tap waterad libitum. Purina is a standard laboratory food contain-ing protein 18%，carbohydrates 50%，fat 3.5%，fibre

6%，calcium 0.8%，phosphorus 0.8%，vitamins (A, D,B 12, K, E, riboflavine, niacin, panthotenic acid, cholinechloride, piridoxine, thiamine, biotin, folic acid) and minerals (copper, Manganese, zinc, iodine and selenium).

2.4 Study protocol

Male rats were divided at random into 3 groups of 18 animals each. Group 1 (Controls) received 2 mI./day of 5%ethanol. Group 2 (low Maca treated) received solution A 2 mI,/day (48 mg/day). Group 3 (high Macatreated) received solution B 2 mL/day (96 mg/day).

Each group was equally divided into 3 subgroupsand they were treated for 7 days, 14 days, and 21 days,respectively. One day after cessation of treatment, therats were sacrificed. The testes and epididymides were removed and cleared off the attached fat and connective tissue. The seminiferous tubules were prepared for transillumination assessment and the epididymis for spermcount.

2.5 Seminiferous tubular assessment

Assessment of the length of stages was made by transillumination under an inverted stereomicroscope at 40x magnification as previously described [5]. For each rat, a total length of 100 cm seminiferous tubules was assessed. The stages assessed included I, II-III, IV-V, VI, VII, VIII, IX-XI, XII and XIII-XIV.

2.6 Epididymal sperm count

Homogenization-resistant epididymal sperm were counted as described previously [6] with some modifi-canons. Homogenization was performed in 5 mL 0.9% saline. Modifications included refrigeration of homogenized epididymal preparation at 4 0C for 24 h to allow sperm be released from the walls. Data are referred as sperm (106) per epididymis.

2.7 Serum testosterone

Serum testosterone levels were determined by radio-immunoassay (RIA) using commercial kit (DiagnosticProducts Co., USA) with IZS-testosterone as the radio-active marker. All samples were run in one assay period.The within assay variation was 5.5%and the sensitivity was 4.0 pg/mL.

2.8 Statistical analysis

Data were presented as meanplusmn;SEM and analyzed using t-test and analysis of variance (one way) when more than two groups were compared. Plt;0.05 was considered significant.

3 Results

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