不同精子质膜保护剂对猪精液常温保存效果的影响

发布时间：2018-04-26 19:34

本文选题：猪精液 + 稀释液　；参考：《西北农林科技大学》2015年硕士论文

【摘要】：为了研究氨基酸类保护剂(牛磺酸、L-谷氨酰胺)和中药活性成分(五味子乙素、三七皂甙R1)对猪精液常温保存效果的影响,首先设计4种稀释液配方,检测其保存过程中精子活率、有效保存时间、质膜完整率、顶体完整率,筛选出保存效果最好的基础配方。然后在基础配方中添加分别添加0.5、1、5、10 mmol/L的牛磺酸(Tau),0.5、1、2、4、8、16 mmo/L的L-谷氨酰胺(Gln),1、5、10、15μmol/L的五味子乙素(Sch B)和0.5、1、2、4、8、16 mmo/L的L-谷氨酰胺(Gln),设置空白对照组,检测精液品质及总抗氧化能力(T-AOC)和丙二醛(MDA)浓度,分析对猪精液保存效果的影响,确定最适添加量,旨在开发新型猪精液常温保存的稀释粉。本研究主要获得如下结果:1.在4种自配猪精液常温保存稀释液中,“稀释液3”,主要成分是葡萄糖40 g/L,柠檬酸钠6.8 g/L,碳酸氢钠0.8 g/L,EDTA1.5 g/L,柠檬酸0.3 g/L,氯化钾0.7 g/L,有效保存时间显著高于其他3种稀释液(P0.05),有效保存时间为4.4 d,保存第4 d的精子活率为0.61,质膜完整率是57.32%,顶体完整率是78.12%;其次是“稀释液2”和“稀释液4”,保存效果最差的是“稀释液1”。因此,“稀释液3”是4种自配猪精液常温保存稀释液中保存效果最好的稀释液配方,可作为基础稀释液。2.在基础稀释液“稀释液3”中分别添加不同浓度的牛磺酸(Tau)和L-谷氨酰胺(Gln),表明添加5 mmol/L的Tau时,精子活率、顶体完整率、总抗氧化能力(T-AOC)浓度显著高于其他组(P0.05),丙二醛(MDA)浓度显著低于其他组(P0.05),保存第5 d时,精子活率为0.66、顶体完整率47.48%、质膜完整率为72.10%、T-AOC浓度4.28 IU/mL、MDA浓度1.63 nmol/mL。添加4 mmol/L的Gln,精子活率、质膜完整率和顶体完整率显著高于其他组(P0.05),保存第5 d时,精子活率为0.59、质膜完整率47.05%、顶体完整率为74.40%、T-AOC浓度4.34 IU/mL、MDA浓度1.51 nmol/mL;添加16 mmol/L的Gln,精子活率、质膜完整率和顶体完整率显著低于其他组(P0.05)。表明添加5 mmol/L的Tau或4 mmol/L的Gln对维持精子活率,保护精子质膜,增强精子抗氧化能力效果显著。3.在“稀释液3”中分别添加不同浓度的五味子乙素(Sch B)和三七皂甙R1(Notoginsenoside R1),表明添加15μmol/L的Sch B精子活率、质膜完整率、T-AOC浓度显著高于其他组(P0.05),MDA浓度显著低于其他组(P0.05),保存第5 d时,精子活率、质膜完整率、T-AOC浓度、MDA浓度分别为0.61、50.71%、4.82 IU/mL、1.45 nmol/mL;添加1、5、10、15μmol/L的Sch B,对精子顶体完整率有明显的保护作用,但组间差异不显著(P0.05)。添加5 mmol/L的三七皂甙R1精子质膜完整率、顶体完整率、T-AOC的浓度显著高于其他组(P0.05),保存第5 d时,精子质膜完整率为57.10%、顶体完整率为73.70%、T-AOC的浓度为4.65 IU/mL;添加1.25、2.5、5、10 mmol/L的三七皂甙R1对降低MDA浓度作用不显著(P0.05);保存第1~3 d,添加1.25、2.5、5、10 mmol/L的三七皂甙R1对降低MDA浓度作用明显,组间有差异性,但差异不显著(P0.05)。因此,Sch B可显著提高精液保存品质,提高猪精液常温保存时抗氧化能力,其最适添加量为15μmol/L。三七皂甙R1可维持精子形态完整,提高猪精液常温保存时抗氧化能力,其最适添加量为5 mmol/L。
[Abstract]:In order to study the effect of amino acid protectant (taurine, L- glutamine) and active ingredients of Chinese medicine (schisandrin B, 37 saponins R1) on the preservation of pig semen at room temperature, first of all 4 kinds of diluent formulations were designed to detect the sperm viability, effective preservation time, the integrity rate of plasma membrane, the integrity of the acrosome, and the best preservation effect. The basic formula was added to the basic formula with 0.5,1,5,10 mmol/L added taurine (Tau), L- glutamine (Gln) of 0.5,1,2,4,8,16 mmo/L, 1,5,10,15 u mol/L schisandin (Sch B) and 0.5,1,2,4,8,16 purified glutamine, and the white control group was set up to detect the quality of semen and the total antioxidant capacity. The effect of two aldehyde (MDA) on the preservation effect of pig semen was analyzed and the optimum addition was determined. The aim of this study was to develop a new type of diluted powder for the preservation of the new pig semen at normal temperature. The main results were as follows: 1. in the diluents of 4 kinds of self matched pig semen, the diluent was 3, the main components were glucose 40 g/L, sodium citrate 6.8 g/L, and sodium bicarbonate 0.8. G/L, EDTA1.5 g/L, 0.3 g/L citric acid and 0.7 g/L of potassium chloride, the effective preservation time is significantly higher than the other 3 diluents (P0.05), the effective preservation time is 4.4 D, the sperm viability of fourth D is 0.61, the integrity rate of the plasma membrane is 57.32%, the integrity rate of the acrosome is 78.12%, and the second is "diluent 2" and "diluent 4", and the worst preservation effect is "dilution". Liquid 1 ". Therefore," diluent 3 "is the best thinning solution for the preservation of 4 kinds of self matched pig semen at normal temperature, which can be used as the base diluent.2. to add different concentrations of taurine (Tau) and L- glutamine (Gln) in the basic diluent" diluent 3 ", indicating that the sperm viability and acrosome are intact when the 5 mmol/L Tau is added. The concentration of total antioxidant capacity (T-AOC) was significantly higher than that of other groups (P0.05), and the concentration of malondialdehyde (MDA) was significantly lower than that of other groups (P0.05). The sperm viability was 0.66, the acrosome integrity rate was 47.48%, the integrity rate of the plasma membrane was 47.48%, the membrane integrity rate was 72.10%, the concentration of T-AOC was 4.28 IU/mL, the concentration of MDA was 1.63 nmol/mL. adding 4 mmol/L Gln, the sperm viability, plasma membrane integrity rate and acrosome. The intact rate was significantly higher than that of the other groups (P0.05). The sperm viability was 0.59, the sperm membrane integrity rate was 47.05%, the integrity rate of the acrosome was 74.40%, the T-AOC concentration was 4.34 IU/mL, and the MDA concentration was 1.51 nmol/mL; the sperm viability, the intact rate of the plasma membrane and the integrity of the acrosome were significantly lower than those of the other groups (P0.05), indicating the Tau or 4 mmol/L to add 5 mmol/L. The effect of Gln on maintaining sperm viability, protecting the sperm plasma membrane and enhancing the antioxidant capacity of sperm was significant.3. in "diluent 3", adding different concentrations of schisandrin B (Sch B) and 37 saponins R1 (Notoginsenoside R1), indicating that the sperm viability of adding 15 u mol/L Sch B, the integrity of plasma membrane and T-AOC concentration were significantly higher than those of the other groups (P0.05). The concentration of the sperm was significantly lower than that of the other groups (P0.05). The sperm viability, plasma membrane integrity, T-AOC concentration, and MDA concentration were 0.61,50.71%, 4.82 IU/mL, 1.45 nmol/mL, respectively, when the preservation of fifth D, and Sch B with 1,5,10,15 micron mol/L had a significant protective effect on the sperm acrosome integrity, but there was no significant difference between the groups (P0.05). The sperm quality was added to the 37 saponins of 5. The membrane integrity, acrosome integrity, T-AOC concentration was significantly higher than that of other groups (P0.05). When the preservation of fifth D, the sperm plasma membrane integrity rate was 57.10%, the acrosome integrity rate was 73.70%, the T-AOC concentration was 4.65 IU/mL, and the 37 saponins R1 added to 1.25,2.5,5,10 mmol/L was not significant (P0.05) for reducing the MDA concentration (P0.05). 1~3 D, added 1~3 D, was added. The effect of 37 saponins R1 on reducing the concentration of MDA was obvious, but the difference was not significant, but the difference was not significant (P0.05). Therefore, Sch B could improve the preservation quality of semen and improve the antioxidant ability of the pig semen at room temperature. The optimum addition amount of 15 mu 37 saponins R1 can maintain the complete morphology of the sperm and improve the antioxidant activity of the pig semen at room temperature. Ability, the optimum amount is 5 mmol/L.

【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828

【参考文献】