Effects of Libyan traditional plants on the reproductive system of male and female rats
Elgenaidi, Abdalla Ramadan
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In different parts of the world, medicinal plants have demonstrated a lot of health benefits to mankind and remains an important source for the discovery of new bio-active compounds. Libya is a typical example of a country where medicinal plants are widely used. Plant extracts of five Libyan medicinal plants were used in this study to investigate their in vivo effects on spermatogenesis and steroidogenesis in male rats and on ovulation and fertility in female rats. The In vitro effects of these plant extracts were also investigated on TM3 Leydig cells and MCF 7 breast cancer cells. A phyto-chemical analysis of the five Libyan medicinal plants (flaxseed, black seeds, radish seed, date palm pollen and nutmeg) was done. The results showed that date palm pollen had a higher antioxidant activity than all of the above mentioned plants. In addition to this, Nigella sativa was observed to possess high flavonol content as well as high antioxidant activity. Male rats exposed to flaxseed, radish seeds and date palm pollen showed no significant alterations in body weight gain, whereas date palm pollen (240 mg/kg, p < 0.05) promoted an increase in body gain. This study also revealed a significant increase in the relative testicular weight of animals exposed to either flaxseed (300mg/kg) or date palm pollen (120mg/kg). In addition, the relative weights of the seminal vesicles of all treated groups showed significant increased values. The level of serum testosterone showed a significant increase after exposure to radish seed (80mg/kg) and a significant dose- dependent increase for date palm pollen when compared to control (P< 0.05). In contrast, flaxseed caused a dose-dependent significant (p <0.01) decrease in testosterone level at radish seed (300mg/Kg). All plant extracts caused a significant increase in sperm concentration. Sperm vitality significantly (p < 0.05) increased by radish seed (80mg/kg), flaxseed (300mg/kg) and date palm pollen (120, 240mg/kg) respectively. Total progressive motility improved significantly at flaxseed (300 mg/kg) (p < 0.001) as well as date palm pollen (p < 0.01). Histological examination of the cross sections of the testis showed clear presence of all stages of spermatogenesis in all the treated groups. Rat epididymides showed normal morphological appearance and their lumen were filled with spermatozoa. The diameter of seminiferous tubules in male rats exposed to date palm pollen (120 and 240 mg/kg) was significantly higher (p < 0.001). The heights of the germ cell epithelia within the eminiferous tubules were also significantly increased in all treated groups. Liver and renal functions tests showed a significant decrease in Alanine transaminase (ALT) and creatinine in all treated groups (p < 0.05), and this demonstrates the lack of cytotoxic effects of date palm pollen, radish seed and flaxseed on the rats. However, these plant extracts produced a non-significant (p > 0.05) increase in Aspartate transaminase (AST) levels. Besides this, superoxide dismutase activity (SOD) in testis was increased significantly by radish seed (160 mg/kg), flaxseed (200 mg/kg) and date palm pollen (120 mg/kg). There was also improved catalase activity in testis of male rats exposed to radish seed and date palm pollen. Regarding male sexual behavior, the time to reach the female and the mount frequency decreased significantly in male rats exposed to flaxseed (300 mg/kg) and date palm pollen (120 and 240 mg/kg; p > 0.05) thus, these plant extracts exhibit aphrodisiac properties. In addition, exposure of male rats to date palm pollen (120 mg/kg) produced a significant (p < 0.01) increase in the number of embryos in untreated female rats. In the female rats, the body weight gain was not affected (p > 0.05). However, the relative uterus weights exposed to nutmeg (200 mg/kg) and date palm pollen (120 and 240 mg/kg) were significantly decreased (p < 0.05). In addition, the relative weights of ovaries after treatment with nutmeg (400 mg/kg) and black seed (400 mg/kg) showed significantly increased values (p < 0.01). Serum FSH was significantly increased (p > 0.05 or 0.01) when the female rats have been exposed to black seed (200 mg/kg), nutmeg (200 mg/kg) or date palm pollen (120 mg/kg). The LH level significantly (p < 0.01) decreased following exposure to black seed (200 mg/kg), date palm pollen (120 mg/kg). On the other hand, serum LH concentration was significantly increased in female rats exposed nutmeg (400 mg/kg; p > 0.05). The creatinine activity in female rat serum in all treated groups was significantly decreased (p < 0.05). Whereas the higher dose of date palm pollen (240 mg/kg) caused only a non-significant decrease. ALT activity in serum of female rat exposed to either black seed (400 mg/kg) or date palm pollen (120 and 240 mg/kg) was shown to decrease significantly (p < 0.05). Histology of the reproductive organs, kidney and liver in the female rats showed no obvious alterations in any of the treated groups. In addition, the number of embryos in female rats significantly increased (p < 0.01; p < 0.001) following exposure of female rats to black seeds 400 and date palm pollen 240 mg/kg, respectively. Incubation of TM3 Leydig cells with radish seeds for 24, 48 or 72 hours caused a significant (p < 0.01) decrease in mitochondrial dehydrogenase activity. Besides that, date palm pollen and flaxseed increased the mitochondrial dehydrogenases activity of TM3 Leydig cells. In addition, higher concentration of date palm pollen, nutmeg and black seed were cytotoxic to MCF7 breast cells. In testis slices testosterone secretion in vitro was significantly increased by flaxseed (500 μg/ml; p > 0·05) and date palm pollen (500 μg/ml; p > 0·01). MCf-7 cells treated with BS 10-50 μg/ml black seed and nutmeg 10-50μg/ml significantly increased cell proliferation. However, the treatment with date palm pollen produced only a weak estrogenic effect, which resulted in a concentration dependent significant increase as observed between 50-1000 μg/ml date palm pollen. In conclusion, in this study, we observed that date palm pollen, radish seed and flaxseed increased libido as well as steroidogenesis and spermatogenesis, improved hepato and nephron-protective effects. In female rats, the plant extracts NM, BS and date palm pollen potentiated the production of gonadotropic hormones. In addition to this, at lower concentrations these medicinal plants promoted cell growth, whereas at higher concentrations they inhibited cell proliferation of MCF- 7 breast cancer cells. The anti-oxidant effects of these plant extracts have been implicated for the above mention effects.