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Synergic effect of pomegranate and maca

extracts against the decrease in sperm

concentration caused by lead acetate

Peralta FJ, Rubio J, Gonzales GF

Faculty of Science and Phylosophy. Department of Biological andPhysiological Sciences. Laboratory of Endocrinology andReproduction. Universidad Peruana Cayetano Heredia. Lima, Peru

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Purpose

The goal of this research was to

determine a synergic effect of theethanolic extract of pomegranate

(Punica granatum) and theaqueous extract of black maca(Lepidium meyenii) alone or in

combination (50%-50%).

We assessed the ability of bothextracts (alone and combined) to

reverse the damage caused bythe administration of lead acetate

(LA) in sperm concentration attestis and epididymis level.

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Materials and Methods

Animals

Thirty six healthy adult male Holtzman rats, at reproductive age

(10 weeks), obtained from the animal house of UniversidadPeruana Cayetano Heredia were used in this study.

Animals were housed six per group and maintained in

polycarbonate cages at environmental temperature (20

22C),humidity between 80% and 85%, and a 12 h light/dark cycle.

Rats were fed Purina laboratory chow and tap water ad libitum.

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Materials and Methods

Preparation of aqueous extract of Maca

First, 500 g of the driedhypocotyls werepulverized and placedin a container with1500 ml of water, and

boiled for 120 min.

Next, the preparationwas left standing tocool and filtered.

Finally, the filtratecontaining dry Macahypocotyls wasplaced in small vialsand kept in 4 Crefrigerator.

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Materials and Methods

Preparation of ethanolic extract of pomegranate

First, seeds wereremoved from the fruit

and discarded.

Then, the pericarp wascut and dried at open-air

shade for 10 days.

After that, it waspulverized and maceratedin ethanol 96C for 8 daysin darkness with constant

agitation.

The extract was filtered,concentrated, andevaporated to constant

weight.

The obtained extract waskept at 4C.

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Materials and Methods

Study design

Group 1: H2Od

Group 2: Lead acetate (LA) 8mg/kg Group 3: LA (8mg/kg) + Ascorbic acid (140mg/Kg)

Group 4: LA (8mg/kg) + Maca (1g/kg)

Group 5: LA (8mg/kg) + Pomegranate (500mg/kg)

Group 6: LA (8mg/kg) and Combined extract (50%-50%)

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Materials and Methods

Treatment

Animals were treated with LA for 35 days. The ascorbic acid and the extractswere administered from day 18 by gavage with an intubation needle N18.

Outcomes

Sperm concentration in epididymis and daily sperm production weredetermined.

Statistical analyses

STATA V.11 was used. If the distribution was normal, differences betweengroups were assessed by analysis of variance (ANOVA). We used theBonferroni test to determine the differences between pair of means.Differences were considered statistically significant when P-value was

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ResultsTable 1.- Percentage of body weight increased in all groups.

GROUPS BODY WEIGHT INCREASED

Control 14%

Lead Acetate (LA) 8%*

LA + Ascorbic Acid 16%+

LA + Maca 15%+

LA + Pomegrante 11%

LA + Combined extract 17%+c

Percentage obtained by the difference between final weight and initial weight

divided by the initial weight. *p

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Results

GROUPS RE LE RT LT LV SV PR

Control 0.580.02 0.560.02 1.760.03 1.760.03 10.460.6 1.010.01 0.410.04

Lead Acetate(LA)

0.490.01* 0.540.01 1.750.03 1.750.03 9.740.11 1.050.06 0.450.02

LA+Ascorbic

Acid

0.580.02+ 0.500.01 1.530.05 1.550.07 11.370.42 1.070.05 0.430.04

LA+Maca 0.510.01 0.510.02 1.700.03 1.640.05 11.540.15 1.050.06 0.400.03

LA+Pomegrante 0.540.01 0.510.01 1.690.03 1.680.05 11.410.11 1.000.06 0.450.04

LA+Combined

Extract

0.540.01 0.550.02 1.660.08 1.700.09 11.691.34 1.050.09 0.410.04

Data are presented as Mean Standard Error. RE: right epididymis, LE: left epididymis, RT: right testicle, LT: left testicle, LV:

liver, SV: seminal vesicle, PR: prostate.*p

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Results

Table 3.- Epididymal count (106

) in all groups.

GROUPS EPIDDYMISCAPUT/CORPUS CAUDA

Control 261.66 3.16 278.42 7.37

Lead Acetate (LA) 208.63 8.97* 208.32 14.9*

LA + Ascorbic Acid 221.85 8.73 209.33 4.5*

LA + Maca 247.75 2.70+ 271.33 8.39+

LA + Pomegrante 301.56 13.76*+ 287.45 19.77+a

LA + Combined extract 234.65 8.74 319.05 18.86+a

Data are presented as Mean Estndar Error. *p

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Results

GROUPS DSP

Control 18.00 0.45

Lead Acetate (LA) 8.23 0.97*LA + Ascorbic Acid 18.94 1.60+

LA + Maca 17.57 0.53+

LA + Pomegrante 19.35 0.61+

LA + Combined extract 22.96 0.20*+a c

Table 4.- Daily Sperm Production (DSP) (106/testicle) in all groups

Data are presented as Mean Standard Error. . *p

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Conclusions

The use of maca and pomegranate separately reverseddeleterious effects of lead acetate in epididymal spermconcentration and daily sperm production.

A combined extract of those plants seems to have abetter effect in reversing the decrease in spermconcentration caused by lead acetate.

This study show evidence of better results by usingmore than one medicinal plant in one extract.

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