本文選題：白藜蘆醇 + 牛精液�。� 參考：《西北農(nóng)林科技大學(xué)》2017年碩士論文

【摘要】：隨著人工授精技術(shù)的快速發(fā)展,精液冷凍保存技術(shù)也不斷得以完善。但是在冷凍保存及解凍過程中,由于諸多因素的存在,解凍后的牛精子的活率、質(zhì)膜完整率、頂體完整率與鮮精相比仍然會大幅度降低,精子受精能力也會大幅度減弱。導(dǎo)致這一問題的原因不僅僅與諸如溫度、滲透壓和氧化應(yīng)激等因素有關(guān),而且可能與精子本身結(jié)構(gòu)及功能的改變有關(guān)。白藜蘆醇是一種天然的非黃酮類多酚類復(fù)合物,在抗炎、抗病毒、抗腫瘤和調(diào)節(jié)氧化還原信號等方面發(fā)揮重要作用,并且能夠與多種細(xì)胞內(nèi)信號通路的靶分子相互作用。目前,關(guān)于不同濃度白藜蘆醇對牛精液冷凍保存效果的影響未見相關(guān)報(bào)道。本研究擬在牛精液冷凍稀釋液中添加不同濃度的白藜蘆醇,測定冷凍后牛精子的活率、頂體完整率、質(zhì)膜完整率及抗氧化指標(biāo)包括總抗氧化能力、丙二醛含量和谷胱甘肽過氧化物酶活力,最后篩選適用于牛精液冷凍保存的白藜蘆醇的最佳添加濃度。本研究主要獲得以下結(jié)果:1.在一定濃度范圍內(nèi)(15~150μM),隨著白藜蘆醇濃度的升高,凍融精子的活率、頂體完整率和質(zhì)膜完整率也隨之升高。但是,當(dāng)白藜蘆醇濃度超過一定水平時(shí),冷凍精子質(zhì)量顯著下降。當(dāng)白藜蘆醇濃度為150μM時(shí),凍融精子的活率、頂體完整率和質(zhì)膜完整率均顯著高于對照組和其他組(P0.05),分別為55.7%、72.5%和53.5%;當(dāng)白藜蘆醇濃度達(dá)到200μM時(shí),精子活率、頂體完整率和質(zhì)膜完整率相對于150μM處理組均有所下降,精子活率顯著低于對照組(P0.05),但質(zhì)膜完整率與對照組相比差異不顯著(P0.05);200μM處理組的頂體完整率與100μM處理組的頂體完整率差異不顯著(P0.05)。表明150μM白藜蘆醇為最適添加濃度。2.在牛精液冷凍保存的過程中,隨著白藜蘆醇添加濃度的升高(15~150μM),精子的T-AOC和GSH-Px活力逐漸增強(qiáng),MDA含量逐漸降低,說明精子的抗氧化能力與白藜蘆醇濃度有一定的劑量依賴性。稀釋液中添加150μM白藜蘆醇,精子的GSH-Px活力最強(qiáng),MDA含量最低,分別達(dá)到113.3μmol/L和96.14 nmol/mL,與對照組差異顯著(P0.05);100μM組精子的T-AOC最高,但與150μM組差異不顯著(P0.05);當(dāng)白藜蘆醇濃度達(dá)到200μM時(shí),其總抗氧化能力(T-AOC)及MDA含量顯著低于對照組(P0.05),GSH-Px活力低于對照組,但差異不顯著(P0.05)。因此,添加150μM白藜蘆醇時(shí),凍融精子的抗氧化能力最高。3.相關(guān)性分析結(jié)果表明,精子活率與T-AOC、MDA含量之間為顯著相關(guān)(P0.05),相關(guān)系數(shù)分別為0.85和-0.76;頂體完整率、MDA含量與T-AOC、GSH-Px的相關(guān)性均不顯著(P0.05);GSH-Px活力與精子活率、質(zhì)膜完整率的相關(guān)性達(dá)到極顯著水平(P0.01),相關(guān)系數(shù)分別為0.99和0.98,這為評估精子膜受損程度提供了一個(gè)新的檢測指標(biāo),該方法更加準(zhǔn)確。綜上所述,在牛精液冷凍稀釋液中添加一定濃度水平的白藜蘆醇能夠提高凍融牛精子的質(zhì)量和抗氧化能力,150μM白藜蘆醇的保護(hù)效果最佳。
[Abstract]:With the rapid development of artificial insemination technology, semen cryopreservation technology has been improved. However, during freezing and thawing, the sperm motility, plasma membrane integrity, acrosome integrity and fertilization ability of the thawed bovine sperm will still be significantly decreased compared with fresh spermatozoa due to the existence of many factors. The causes of this problem are not only related to factors such as temperature, osmotic pressure and oxidative stress, but also to changes in the structure and function of sperm itself. Resveratrol is a natural non-flavonoid polyphenol complex, which plays an important role in anti-inflammatory, anti-virus, anti-tumor and redox signal regulation, and can interact with target molecules of various intracellular signaling pathways. At present, there is no report on the effect of resveratrol at different concentrations on freezing preservation of bovine semen. In this study, different concentrations of resveratrol were added to frozen bovine semen to determine the sperm motility, acrosome integrity, plasma membrane integrity and antioxidant indexes including total antioxidant capacity. Malondialdehyde (MDA) content and glutathione peroxidase (GSH) activity were determined. Finally, the optimal concentration of resveratrol for frozen preservation of bovine semen was selected. In this study, the following results were obtained: 1: 1. With the increase of resveratrol concentration, the motility, acrosome integrity and plasma membrane integrity of freeze-thawed spermatozoa increased within a certain concentration range (15 ~ 150 渭 M). However, when the concentration of resveratrol exceeded a certain level, the quality of frozen sperm decreased significantly. When the concentration of resveratrol was 150 渭 M, the viability rate, acrosome integrity rate and plasma membrane integrity rate of freeze-thawed sperm were significantly higher than those of the control group and other groups (P0.05), which were 55.7% and 53.5%, respectively, when the concentration of resveratrol reached 200 渭 M, the sperm motility rate was significantly higher than that of the control group and other groups. The acrosome integrity rate and plasma membrane integrity rate decreased compared with 150 渭 M treatment group. Sperm motility was significantly lower than the control group (P0.05), but the plasma membrane integrity rate was not significantly different from the control group (P0.05). The acrosome integrity rate of 200 渭 M treatment group was not significantly different from that of 100 渭 M treatment group (P0.05). The optimum concentration of resveratrol was 150 渭 M. During the cryopreservation of bovine semen, with the increase of resveratrol concentration (15 ~ 150 渭 M), the activity of T-AOC and GSH-Px decreased gradually, indicating that the antioxidant ability of sperm was dependent on the concentration of resveratrol in a dose-dependent manner. The concentration of GSH-Px in sperm was the lowest, reaching 113.3 渭 mol / L and 96.14 nmol / mL, respectively. The T-AOC of spermatozoa was the highest in 100 渭 M group (P0.05), but not significantly different from that in 150 渭 M group (P0.05), and when the concentration of resveratrol reached 113.3 渭 M, the concentration of resveratrol reached 200 渭 M. The total antioxidant capacity (T-AOC) and MDA content were significantly lower than the control group (P0.05) GSH-Px activity was lower than the control group, but the difference was not significant (P0.05). Therefore, when 150 渭 M resveratrol was added, the antioxidation ability of freeze-thawed spermatozoa was the highest. 3. The results of correlation analysis showed that there was a significant correlation between sperm motility and T-AOCX MDA content (P0.05), the correlation coefficients were 0.85 and -0.76, respectively, and the correlation between acrosome integrity rate and T-AOCnGSH-Px activity was not significant (P0.05), and there was no significant correlation between sperm motility and sperm motility. The correlation of plasma membrane integrity rate was very significant (P0.01), the correlation coefficients were 0.99 and 0.98, respectively, which provided a new index for evaluating the degree of sperm membrane damage, and the method was more accurate. In conclusion, adding a certain concentration of resveratrol to the frozen diluent of bovine semen can improve the quality of frozen and thawed bovine spermatozoa and the antioxidant ability of 150 渭 M resveratrol.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S823

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