돼지 정자의 체외보존에 있어 Pyruvate, Taurine 및 Melatonin의 항산화효과와 평가방법의 상관관계 분석
Antioxidative Effect of Pyruvate, Taurine and Melatonin, and Relationships of the Evaluation Methods on Boar Sperm Viability for In Vitro Storage
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본 연구는 돼지 정액의 체외보존에 있어 pyruvate, taurine 및 melatonin의 항산화 효과와 생존력의 평가방법간의 상관관계를 검토하고자 실시하였다. 돼지 정액의 체외보존시 항산화 효과를 평가하기 위하여 체외보존 신선정액에 pyruvate(1mM), taurine(50 mM) 및 melatonin(100nM)에 H2O2(100uM)을 혼합첨가 및 항산화제 단독처리하여 체외에서 3~12시간까지 배양하면서 3시간 간격으로 정자의 운동성, 정자의 원형질막의 완전성 및 지질 산화도를 측정하였다. 정자의 운동성은 항산화제 첨가구가 다른 처리구에 비해 높은 결과를 나타내었고, Hypoosmotic swelling test(HOST) 방법을 이용한 정자의 원형질막 완전성에서도 운동성과 유사한 결과를 나타내었다. 정자의 원형질막의 지질산화도 검사에서는 항산화제 첨가구와 무첨가구가 H2O2 첨가구와 H2O2와 항산화제의 혼합 첨가구에 비해 낮은 결과를 나타내었다. 또한 평가방법들 간의 상관관계에서는 운동성과 정자의 원형질막의 완전성에는 정의 상관관계를 나타내었고, 정액의 성상검사 방법과 지질산화검사 방법에서는 부의 상관관계를 나타내었다. 본 실험의 결과는 정액희석액에 항산화제의 첨가는 정자의 운동성 및 정자의 원형질막 완전성을 증가시키고, 정자의 원형질막의 지질산화도를 감소시키는 것으로 나타났다. 또한 정자의 원형질막의 완전성 검사 (HOST)는 정자의 원형질막의 지질산화도 검사 및 운동성 검사를 대신하여 정자의 질적 평가에 유용하게 이용될 수 있을 것으로 사료된다.
Abstract
The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(－0.23~－0.92 and －0.68~－0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.
참고문헌 (19)
- Aitken, "A free radical theory of male infertility"
- Aitken, R. J., "Free radicals", 7, pp.659~668 (1995)
- Aitken, R. J., "Aitken, R. J. and Krausz, C.", 122, pp.497~506 (2001), Reproduction
- Ball, B. A.,, "Effect of antioxidants on preservation of motility viability and acrosomal integrity of equine spermatozoa during storage at 5 degrees C", 56, pp.577~589 (2001)
- Cerolini, S.,, "Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage", 58, pp.99~111 (2000)
- Chen, "Hydroxyl radical-induced decline in motility and increase in lipid peroxidation and DNA modification in human sperm Effects of hydrogen peroxide on DNA and plasma membrane integrity of human spermatozoa", pp.291 1200~303 1207
- and Sano, "Select anti- oxidants improve the function of extended boar semen stored at 10 degrees C lipid peroxidation and enzymatic defence systems in human spermatozoa", therioge(logy 63), pp.1605 17~1616 26
- A, "Assessment of sperm characteristics post-thaw and response to calcium ionophore in relation to fertility in Swedish dairy AI bulls", therioge(gy 53), pp.859~875 (2000)
- Van der Ven, "Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics", pp.219~228 (1984)
- de Lamirande, "Impact of reactive oxygen species on spermatozoa a balancing act between beneficial and detrimental effects", pp.15~21 (1995)
- Raes, "Importance of Se-glutathione peroxidase and Cu", Zn-SOD for cell survival against oxidative stress, 1994, 235~248
- Morales, "Pyruvate prevents peroxide-induced injury of in vitro preimplantation bovine embryos", pp.149~157 (1999)
- Nie, "Adaptation of the hypoosmotic swelling test to assess functional integrity of stallion spermatozoal plasma membranes", therioge(logy 55), pp.1005~1018 (2001)
- Ogasawara, "Reactivity of taurine with aldehydes and its physiological role", pp.2172~2175 (1993)
- Lorenzo, "A short hypo- osmotic swelling test for the prediction of boar sperm fertility", perez-lla therioge(logy 56), pp.387~398 (2001)
- Sharma, "Role of reactive oxygen species in male infertility", pp.835~48 50 (1996)
- "Relative impact of oxidative stress on male reproductive function", pp.851~862 (2001)
- Twigg, "Oxidative damage to DNA in human spermatozoa does not preclude pronucleus formation at intracytoplasmic sperm injection"
- White, "Lipids and calcium uptake of sperm in relation to cold shock and preservation", pp.639~658 (1993)