Potency of Inhibin B as Candidates Male Hormonal Contraception based on Sperm Quality, Expression of cAMP Response Element Modulator (CREM) in testis and Level of Serum LH in Rattus norvegicus
Large population, in addition to being capital for the country's development is also contrary to the problem and burden for the country. One effort to control increasing population is contraception. Men should also have the same burden in family planning, so that male contraceptive methods should be developed. One of targets of male contraception is disturbing production of spermatozoa which is regulated by FSH and LH. FSH is required for production and development of gametes in fertile phase of life. Secretion and production of FSH by pituitary gland in males is regulated by inhibin B, which produced by Sertoli cells in testis. Inhibin B is a polypeptide hormone that selectively inhibits FSH secretion via negative feedback mechanism, so that inhibin B has potential as one of candidates for male contraception. Free of side effects and especially without influence on masculinity and libido was required in an ideal male contraceptive method.
The main signaling pathway of FSH in Sertoli cells is cAMP-PKA pathway that regulate the transcription factor CREM. CREM is a transcription factor that plays an important role in spermatogenesis. The purpose of this study was to determine the role of inhibin B isolate from testis primary culture in quality of spermatozoa, expression of CREM in testis tissue and levels of serum LH in rats (Rattus norvegicus) strain Wistar. The quality of spermatozoa based on motility, viability and concentration of spermatozoa. Research procedures divided into two stage, (1) production and characterization of inhibin B, includes testis primary culture of Rattus norvegicus, isolation of crude protein from rat testis primary culture, separation of the crude protein by native PAGE, test specificity by western blotting, and inhibin B levels measured by ELISA, (2) treatment of animals model and test potency of inhibin B as a candidate male contraceptive include inhibin B isolates administration into rat, sperm quality test, measurement of serum LH levels by ELISA method, test CREM expression by immunohistochemistry. Data expression of CREM in testis tissue was analyzed by descriptive analysis, whereas sperm concentration, sperm motility, sperm viability and LH levels were analyzed using ANOVA.
The results showed that inhibin B from primary culture of rat testis has a molecular weight of 28 and 30 kDa based on result of western blotting with antibodies anti-rat inhibin βB. CREM expression in treated group of inhibin B was detected in interstitial cells but CREM expression in the tubuli seminiferous decreased. All treated group of inhibin B showed a significant decrease (P <0.05) of concentration, sperm motility and viability compared with control. Concentration of spermatozoa in group of KI, K II and K III, respectively, 3.54 ± 0.05, 3.44 ± 0.08 and 3.15 ± 0.09 million / mL and lower than control (K0) (3.7 ± 0.05 million / mL). Motility of spermatozoa in group of KI, K II and K III, respectively 44.83% ± 1.35, 39.5% and 30.5% ± 1.75 ± 1.43 and lower than control (69 , 17% ± 0.94). Viability of spermatozoa in group of KI, K II and K III, respectively 46.33% ± 1.14, 40.83% ± 1.58 and 26.66% ± 1.33 and lower than control (70 , 5% ± 3.05). Serum LH levels in rats after administration of inhibin B showed no significant differences (P <0.05) between control and treated group. Based on these results, it can be concluded that inhibin B has potential as a candidate male contraception because it can induce the decrease in sperm quality and expression of CREM in testis, but it also had no impact on LH that regulate masculinity and libido.
Keywords : contraception, CREM, inhibin B, quality of spermatozoa, LH.
The main signaling pathway of FSH in Sertoli cells is cAMP-PKA pathway that regulate the transcription factor CREM. CREM is a transcription factor that plays an important role in spermatogenesis. The purpose of this study was to determine the role of inhibin B isolate from testis primary culture in quality of spermatozoa, expression of CREM in testis tissue and levels of serum LH in rats (Rattus norvegicus) strain Wistar. The quality of spermatozoa based on motility, viability and concentration of spermatozoa. Research procedures divided into two stage, (1) production and characterization of inhibin B, includes testis primary culture of Rattus norvegicus, isolation of crude protein from rat testis primary culture, separation of the crude protein by native PAGE, test specificity by western blotting, and inhibin B levels measured by ELISA, (2) treatment of animals model and test potency of inhibin B as a candidate male contraceptive include inhibin B isolates administration into rat, sperm quality test, measurement of serum LH levels by ELISA method, test CREM expression by immunohistochemistry. Data expression of CREM in testis tissue was analyzed by descriptive analysis, whereas sperm concentration, sperm motility, sperm viability and LH levels were analyzed using ANOVA.
The results showed that inhibin B from primary culture of rat testis has a molecular weight of 28 and 30 kDa based on result of western blotting with antibodies anti-rat inhibin βB. CREM expression in treated group of inhibin B was detected in interstitial cells but CREM expression in the tubuli seminiferous decreased. All treated group of inhibin B showed a significant decrease (P <0.05) of concentration, sperm motility and viability compared with control. Concentration of spermatozoa in group of KI, K II and K III, respectively, 3.54 ± 0.05, 3.44 ± 0.08 and 3.15 ± 0.09 million / mL and lower than control (K0) (3.7 ± 0.05 million / mL). Motility of spermatozoa in group of KI, K II and K III, respectively 44.83% ± 1.35, 39.5% and 30.5% ± 1.75 ± 1.43 and lower than control (69 , 17% ± 0.94). Viability of spermatozoa in group of KI, K II and K III, respectively 46.33% ± 1.14, 40.83% ± 1.58 and 26.66% ± 1.33 and lower than control (70 , 5% ± 3.05). Serum LH levels in rats after administration of inhibin B showed no significant differences (P <0.05) between control and treated group. Based on these results, it can be concluded that inhibin B has potential as a candidate male contraception because it can induce the decrease in sperm quality and expression of CREM in testis, but it also had no impact on LH that regulate masculinity and libido.
Keywords : contraception, CREM, inhibin B, quality of spermatozoa, LH.
.jpg)
No comments