大豆卵磷脂在人类精子超低温冷冻保存中的应用研究

doi:10.12280/gjszjk.20250524

摘要/Abstract

摘要：

目的：基于HSPM保护剂的无动物源性冷冻体系，探究大豆卵磷脂（soybean lecithin，SL）在人类精子超低温冷冻保存中的应用。方法：收集156例男性精液样本（正常精子92例，弱活力精子64例），以HSPM+20%卵黄（EY组）为对照组，设置1%、2%、3%、5% SL（SL组）为实验组。分析精子活力、DNA碎片指数（DNA fragmentation index，DFI）、精子形态、细胞膜完整性及氧化应激水平。结果：在正常精子和弱活力精子中，精子活力方面，2% SL组与EY组的前向运动（progressive motility，PR）、平均路径速度（average path velocity，VAP）、曲线速度（curvilinear velocity，VCL）和总活力（total sperm motility，TM）显著高于其余3组（均P<0.05），2% SL组与EY组的PR、VAP差异无统计学意义（均P>0.05）；膜完整性方面，2% SL组与EY组低渗肿胀试验阳性率、活细胞率显著高于其余3组（均P<0.05），2% SL组与EY组差异无统计学意义（均P>0.05）；DFI、正常精子形态率及氧化应激水平方面，2% SL组显著优于其余4组（均P<0.05）。结论：2% SL可替代EY用于HSPM保护剂，在维持精子活力与膜完整性的同时，显著改善DNA稳定性、形态完整性并降低氧化损伤。

关键词: 精子, 卵磷脂类, 精液保存, 低温保存, 精子能动性

Abstract:

Objective: To evaluate the cryoprotective effects of soybean lecithin (SL) in an animal-free cryopreservation system based on HSPM cryoprotectant for human sperm cryopreservation. Methods: A total of 156 human semen samples (92 normal sperm and 64 sperm with weak motility) were collected. Using HSPM+20% egg yolk (EY group) as the control group, the experimental groups were set up with 1%, 2%, 3%, and 5% SL (SL groups). Sperm motility, DNA fragmentation index (DFI), sperm morphology, membrane integrity, and oxidative stress level were analyzed. Results: In terms of sperm activity, the 2% SL group and EY group showed significantly higher progressive motility (PR), average path velocity (VAP), curvilinear velocity (VCL), and total sperm motility (TM) in the normal sperm and weakly motile sperm when compared to the other three groups (all P<0.05). However, there was no statistically significant difference in PR and VAP between the 2% SL group and EY group (both P>0.05). In terms of membrane integrity, the positive rate of hypoosmotic swelling test and viability in the 2% SL group and EY group were significantly higher than those in the other three groups (all P<0.05), while there were no statistically significant difference between the 2% SL group and EY group (all P>0.05). In terms of DFI, normal sperm morphology rate and oxidative stress level, the 2% SL group was significantly superior to the other four groups (all P<0.05). Conclusions: The 2% SL formulation can effectively replace EY in the HSPM cryoprotectant. It maintains sperm motility and membrane integrity, while significantly improves DNA stability, morphological integrity and reduces oxidative damage.

Key words: Spermatozoa, Lecithins, Semen preservation, Cryopreservation, Sperm motility

李欢, 陈伊, 蒋祥龙, 陈胜辉, 熊青. 大豆卵磷脂在人类精子超低温冷冻保存中的应用研究[J]. 国际生殖健康/计划生育杂志, 2026, 45(1): 6-10.

LI Huan, CHEN Yi, JIANG Xiang-long, CHEN Sheng-hui, XIONG Qing. Application of Soybean Lecithin in the Cryopreservation of Human Sperm[J]. Journal of International Reproductive Health/Family Planning, 2026, 45(1): 6-10.

图/表 5

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精子类别	n	精子浓度（×10⁶/mL）	PR （%）	VAP （μm/s）	VCL （μm/s）	TM （%）
正常精子	92	69.5±25.2	46.8±7.8	38.3±7.2	65.0±8.1	57.5±8.9
弱活力精子	64	43.8±15.6	22.7±6.9	26.0±5.8	45.4±7.9	29.9±6.7
t		7.847	19.858	11.881	14.948	20.977
P		<0.001	<0.001	<0.001	<0.001	<0.001

精子类别	n	精子浓度（×10⁶/mL）	PR （%）	VAP （μm/s）	VCL （μm/s）	TM （%）
正常精子	92	69.5±25.2	46.8±7.8	38.3±7.2	65.0±8.1	57.5±8.9
弱活力精子	64	43.8±15.6	22.7±6.9	26.0±5.8	45.4±7.9	29.9±6.7
t		7.847	19.858	11.881	14.948	20.977
P		<0.001	<0.001	<0.001	<0.001	<0.001

组别	正常精子（n=92）				弱活力精子（n=64）
组别	PR（%）	VAP（μm/s）	VCL（μm/s）	TM（%）	PR（%）	VAP（μm/s）	VCL（μm/s）	TM（%）
EY组	41.9±5.1	39.0±4.3	67.9±5.8	56.5±6.8	20.4±3.2	24.8±4.0	43.3±5.6	29.1±3.8
1% SL组	36.2±5.1^ab	33.3±4.0^ab	58.1±4.6^ab	49.1±5.8^ab	16.8±3.2^ab	20.4±2.9^ab	36.6±4.8^ab	24.3±3.0^ab
2% SL组	42.1±3.6	38.1±4.3	65.6±5.4^a	56.0±3.1	21.4±3.1	24.8±3.8	41.0±5.1^a	27.6±4.0^a
3% SL组	33.6±3.9^ab	30.1±3.8^ab	52.1±5.0^ab	45.8±5.7^ab	14.5±3.0^ab	18.2±3.0^ab	34.2±4.2^ab	20.9±3.2^ab
5% SL组	28.9±4.5^ab	28.1±3.7^ab	50.3±3.6^ab	42.6±5.5^ab	12.9±2.8^ab	16.0±3.3^ab	33.0±4.3^ab	19.1±3.2^ab
F	145.464	131.146	236.675	115.356	89.657	83.353	53.805	96.890
P	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001

组别	正常精子（n=92）				弱活力精子（n=64）
组别	PR（%）	VAP（μm/s）	VCL（μm/s）	TM（%）	PR（%）	VAP（μm/s）	VCL（μm/s）	TM（%）
EY组	41.9±5.1	39.0±4.3	67.9±5.8	56.5±6.8	20.4±3.2	24.8±4.0	43.3±5.6	29.1±3.8
1% SL组	36.2±5.1^ab	33.3±4.0^ab	58.1±4.6^ab	49.1±5.8^ab	16.8±3.2^ab	20.4±2.9^ab	36.6±4.8^ab	24.3±3.0^ab
2% SL组	42.1±3.6	38.1±4.3	65.6±5.4^a	56.0±3.1	21.4±3.1	24.8±3.8	41.0±5.1^a	27.6±4.0^a
3% SL组	33.6±3.9^ab	30.1±3.8^ab	52.1±5.0^ab	45.8±5.7^ab	14.5±3.0^ab	18.2±3.0^ab	34.2±4.2^ab	20.9±3.2^ab
5% SL组	28.9±4.5^ab	28.1±3.7^ab	50.3±3.6^ab	42.6±5.5^ab	12.9±2.8^ab	16.0±3.3^ab	33.0±4.3^ab	19.1±3.2^ab
F	145.464	131.146	236.675	115.356	89.657	83.353	53.805	96.890
P	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001	<0.001

组别	正常精子（n=92）		弱活力精子（n=64）
组别	DFI （%）	正常精子形态率（%）	DFI （%）	正常精子形态率（%）
EY组	22.0±2.7^a	38.2±4.0^a	27.7±2.2^a	33.0±2.9^a
1% SL组	24.1±2.6^a	37.4±4.4^a	28.4±2.3^a	30.3±4.0^a
2% SL组	16.7±1.8	45.3±3.9	24.6±2.1	39.0±4.1
3% SL组	26.2±1.9^a	36.1±3.8^a	28.3±2.1^a	28.4±4.3^a
5% SL组	29.1±2.6^a	31.4±2.9^a	31.2±2.2^a	28.2±3.3^a
F	371.022	156.699	72.553	90.550
P	<0.001	<0.001	<0.001	<0.001