Philosophiae Doctor - PhD / Camellia sinensis (tea) and Aspalathus linearis (rooibos) may improve reproductive function owing to their antioxidant properties. To test this hypothesis, male and female rats were given 2% and 5% green tea (Gt), black tea (Bt), unfermented rooibos (Ur) or fermented rooibos (Fr) as sole source of drinking for 52 and 21 days respectively. Control rats received tap water. In addition, TM3 Leydig cells were exposed to 0.025, 0.05, 0.1 and 0.5 % aqueous extracts of green tea, black tea, unfermented and fermented rooibos for 24h. In vitro analysis of tea and the herbal infusion revealed the phenolic property and antioxidant capacity (FRAP) in the order Gt > Bt > Ur > Fr. Camellia sinensis and Aspalathus linearis revealed no significant effect on serum antioxidant capacity (p > 0.05) and lipid peroxidation (MDA) in the kidney or liver in both male and female rats and in the testes of the male rats (p > 0.05). In addition, the antioxidant levels were maintained in the testes, liver and kidneys in both the male and female rats. In the male rats, no significant alterations were observed in body weight gain, liver and reproductive organs weight, and serum testosterone (p > 0.05). Only, 5% green tea significantly increased testosterone level (p < 0.05). Seminiferous tubules displayed complete spermatogenesis with abundant sperm in the lumen in all treated groups. However, a significant decrease in diameter and germinal epithelial height of these tubules were observed (p < 0.05). In the epididymides, epithelial height of caput region showed a significant increase (p < 0.01), while the cauda region was increased by Camellia sinensis but decreased by Aspalathus linearis. Sperm concentration improved significantly by green tea and unfermented rooibos (p < 0.05), while black tea and fermented rooibos produced a non significant effect (p > 0.05). Sperm viability was enhanced in all treatment groups (p < 0.05). Furthermore, green tea, black tea and unfermented rooibos significantly improved the motility of rat sperm (p < 0.05); fermented rooibos tended to improve it (p > 0.05). In addition, green tea, black tea and fermented rooibos enhanced acrosome reaction (p < 0.05). Creatinine activity was significantly higher in rats treated with black tea, unfermented rooibos or fermented rooibos (p < 0.05), green tea tended to increase it (p > 0.05) reflecting the significant increased kidney weight in the treatment groups at high concentrations. Liver markers, ALT and AST, decreased significantly in all treated groups (p < 0.05), except in 5% fermented rooibos where a significant increase in AST level was observed (p < 0.01). In the female rats, the body weight gain, and reproductive organs weight was no affected (p > 0.05). However, 5% fermented rooibos reduced the ovarian weight (p < 0.05), while 5% unfermented rooibos significantly increased the uterine weight (p < 0.05). Liver weight increased significantly by black tea and unfermented rooibos (p < 0.05) while the kidney weight increased significantly by 5% black tea (p < 0.05). No significant effect was observed in the level of FSH produced, on the other hand, Camellia sinensis significantly lowered the level of LH (p < 0.05), while Aspalathus linearis had no effect (p > 0.05). Creatinine activity was enhanced significantly only by 5% fermented rooibos (p < 0.05). Liver markers, ALT and AST were reduced in most treated groups except in fermented rooibos where an increase was observed. In addition, histological sections revealed no obvious alteration in the ovaries, uteri, kidneys and liver of all treated female rats. Camellia sinensis and Aspalathus linearis significantly reduced the level of testosterone produced in TM3 Leydig cells under stimulated conditions in vitro (p< 0.05). Furthermore, both plants maintained the viability and morphology of the cells. However, at 0.5% of either plant extracts, a significant decrease in the viability (p < 0.05) and altered morphology of the TM3 Leydig cells was observed. In conclusion, Camellia sinensis and Aspalathus linearis significantly improved certain sperm function which might be attributed to their high level of antioxidant activity. However, the prolonged exposure of both plant extracts might result in subtle structural changes in the male reproductive system and impair kidney function. In addition, fermented rooibos at high concentration may also impair the functions of the liver. In vitro, both plants were shown to possess anti-androgenic property on TM3 Leydig cells. Furthermore, both Camellia sinensis and Aspalathus linearis may be classified as weak phytoestrogens due to the changes in the weight of the uterus and ovaries observed. / South Africa

Philosophiae Doctor - PhD / 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.

Thesis (M.Sc. (Medical Sciences)) -- University of Limpopo, 2020 / Infertility affects nearly 186 million couples globally, with male factors contributing to half of the cases. Oxidative stress is an established cause of declining semen quality. Moringa oleifera has proven antioxidants. This study aimed to investigate in vitro effects of aqueous leaf extract of M. oleifera on human sperm functions. Semen samples from donors (n = 40) and patients (n = 30) were washed with HTF-bovine serum albumin (BSA), and then incubated with various concentrations of M. oleifera (0, 0.625, 6.25, 62.5, and 625 μg/ml) at 37°C for 1 hour. Sperm motility, vitality, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), DNA fragmentation, capacitation, and acrosome reaction were assessed. Sperm motility, vitality, MMP, and capacitation were enhanced, while ROS production, and DNA fragmentation decreased after M. oleifera treatment. Uncapacitated spermatozoa increased significantly with a reduction in acrosome reaction in donors. M. oleifera antioxidant compounds suppressed excessive ROS, preserved mitochondrial membrane, DNA and acrosome integrity, while enhancing sperm motility and viability. / National Research Foundation (NRF)

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-09T12:03:21Z No. of bitstreams: 1 Flavia Camila Siqueira Pereira Ramalho.pdf: 838412 bytes, checksum: e92375da22d319dd80763723a0bdbfbf (MD5) / Made available in DSpace on 2017-02-09T12:03:21Z (GMT). No. of bitstreams: 1 Flavia Camila Siqueira Pereira Ramalho.pdf: 838412 bytes, checksum: e92375da22d319dd80763723a0bdbfbf (MD5) Previous issue date: 2015-02-09 / In vitro embryo production (IVP) in cattle became an important commercial tool in genetic improvement programs of the world herd, being widely used for this purpose. However in vitro fertilization (IVF) can cause generation of reactive oxygen species that can affect embryo viability. Coenzyme Q10, an important cofactor in the transport chain of mitochondria has antioxidant function on lipid membrane and it was proved a direct correlation between the presence of Coenzyme Q10 and normal spermatozoa parameters such as density, motility, morphology and volume. The objective of this study was to evaluate the effect of Coenzyme Q10 on sperm function in IVF using conventional or sexed semen; also if the addition of this cofactor can improve embryo IVP in bovine oocytes. In experiment 1 was evaluated the effect of sperm function during incubation periods of sexed and conventional semen samples. In experiment 2, IVF medium was supplemented with 0 (control group), 5 μM, 10 μM, 20 μM of Coenzyme Q10. Bovine oocytes were collected from a slaughterhouse located 20 minutes from the lab. It was observed a negative effect of Coenzyme with significant differences in the rates of cleavage or in the production of blastocyst (p< 0.05) at a concentration of 20 μM when compared with all other groups with either sexed as conventional semen. These results demonstrate that supplementation of the Coenzyme Q10 in the IVF medium, do not alter spermatozoa function. We can also infer that there is a tendency to improve embryo production in the concentration of 5 μM in IVF medium. / A produção de embriões in vitro (PIV) em bovinos tornou-se importante ferramenta comercial nos programas de melhoramento genético do rebanho mundial como técnica de multiplicação, sendo amplamente utilizada para esse fim. Entretanto a Fertilização in vitro (FIV) provoca geração de espécies reativas de oxigênio que podem afetar a viabilidade embrionária. A Coenzima Q10, um cofator de importância na cadeia de transporte das mitocôndrias tem função antioxidante na membrana lipídica e foi verificado uma correlação direta entre a Coenzima e os parâmetros normais de sêmen tais como, densidade, motilidade, morfologia e o volume. O objetivo desse trabalho foi avaliar o efeito da Coenzima Q10 na função espermática em FIV utilizando-se sêmen convencional e sexado; e se a adição desse cofator pode melhorar a produção embrionária in vitro de oócitos bovinos. No Experimento o meio de FIV foi suplementado com 0 (grupo controle), 5 μM, 10 μM, 20 μM da Coenzima Q10. Os Oócitos foram coletados de um abatedouro localizado a 20 minutos do laboratório. Foi observado um efeito deletério da coenzima com diferença significativa nas taxas de clivagem ou na produção de blastocisto (p<0,05) na concentração de 20 μM quando comparado com os demais grupos tanto com sêmen sexado como convencional. Estes resultados demonstram que a suplementação da Coenzima Q10 no meio FIV não altera a função espermática, entretanto tem um efeito deletério a partir da concentração de 20 μM. Podemos ainda inferir que na concentração de 5 μM no meio FIV há uma tendência na melhoria da produção embrionária.