EFFECT OF ETHANOL EXTRACT PUMPKIN (CUCURBITA PEPO) SEEDS ON BALB/C MALE MICE FERTILITY

• Main Author: Solihah, Hani
• Format: Theses
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/70353
• Institution: Institut Teknologi Bandung

The gap in contraceptive use between men and women due to the limited methods of contraception for men is the basis for the importance of conducting studies that can provide references for the development of new contraceptive methods for men. In addition, contraceptive methods for males that are more effective and do not conflict with animal welfare principles still need to be developed. It is known that 577 plant species in various families have active compounds that are antifertility, but the use of antifertility compounds derived from natural ingredients is still very limited, especially in Indonesia. Cucurbita is known to be rich in phytosterol compounds. Phytosterols from Cucurbita have been widely used as compounds for prostate cancer therapy because they are proven to affect androgen hormon regulation in men, including inhibiting 5-alpha reductase 2 (5AR-2) activity. Currently there are not many studies reporting the effect of C. pepo extract which has inhibitory activity on 5AR-2 on fertility, especially sperm quality and serum testosteron, thus the potential of C. pepo as a plant candidate for antifertility therapy in men is unknown. Based on this, the researchers wanted to see how the effect of the ethanol extract of C. pepo seeds rich in phytosterols on the fertility factors of male BALB/c strain mice. Phytochemical test of C. pepo seed extract was carried out by qualitative test and GC-MS methode. The toxicity test to determine the LD50 value of C. pepo seed extract was carried out using the Acute Toxic Class 423 method in accordance with the testing guidelines of The Organization for Economic Co-operation and Development (OECD). The inhibition activity of C. pepo extract against the enzyme 5-alpha reductase 2 was carried out through the HPLC method by observing the decrease in hydrolyzed substrate. Fertility factors were analyzed by measuring serum testosteron levels using the competitive ELISA method; calculation of the Gonadosomatic Index value; spermatozoa viability; spermatozoa motility; and sperm concentration. The values of sperm motility, viability, and concentration were calculated by observing the sperm suspension from the epididymis and observed using microscope with a magnification of 400x in ten fields of view. Phytochemical analysis showed that C. pepo seed extract contained steroids, terpenoids, flavonoids, alkaloids, and was rich in various phytosterol compounds. C. pepo seed extract is categorized as a mild toxic compound with an LD50 cut off value of 2500 mg/kgBB. The results of the 5AR-2 activity test by analyzing the results of 5AR-2 substrate quantification with HPLC showed a decrease in 5AR-2 substrate hydrolysis in BALB/c mice fed C. pepo seed extract doses of 200 mg/kgBW and 600 mg/kgBW compared to the control group with activity values 7.153x10^(-8) units/mg each; 5.619x10^(-8) units/mg; and 1.837x10^(-8) units/mg (p<0.05). The results of the ELISA test showed an increase in serum testosteron levels in BALB/c mice given C. pepo extract relative to controls with testosteron levels of 4.161 ng/ml each; 13,591ng/ml; and 11.675 ng/ml (p<0.05). Administration of C. pepo seed extract significantly reduced prostate and seminal vesicle weights with GSI values of 1.437% respectively; 1.137%; 1.105%. In addition, the results of the study showed that the administration of C pepo seed extract significantly reduced sperm concentration with respective values of 72.429x10^6 million/ml; 46,714x10^6 million/ml; and 31.143x10^6 million/ml, and decreased sperm viability with a viability percentage of 64.29% respectively; 20%; and 21% (p<0.05). The administration of C. pepo seed extract showed a decrease in motility in the 200 mg/kgBB and 600 mg/kgBB dose groups compared to the control group with a percent motility value of 38.503% respectively; 28.511 %; and 28.708% (p>0.05). There are indications that C. pepo seed extract has inhibitory activity against 5-alpha reductase 2 thereby reducing the process of testosteron catabolism, affecting the weight of organs that depend on androgen hormons such as the prostate and seminal vesicles, and affecting the sperm quality of male BALB/c stain mice.