基于拉曼光谱分析猪卵母细胞玻璃化冷冻前后变化

doi:10.11843/j.issn.0366-6964.2025.10.020

摘要/Abstract

摘要：

旨在应用拉曼光谱技术研究猪体外成熟卵母细胞玻璃化冷冻前后分子结构的变化。本研究采集屠宰场猪离体卵巢，经实验室抽取卵丘-卵母细胞复合体，体外成熟培养后，分别收集新鲜卵母细胞和玻璃化冷冻-解冻卵母细胞，采用激光共焦拉曼光谱仪获取两组卵母细胞的拉曼光谱信息，对数据进行质控后，分析玻璃化冷冻前后卵母细胞的分子结构变化。结果表明，与新鲜组卵母细胞相比，玻璃化冷冻-解冻组卵母细胞在1 020~1 140 cm^-1的碳水化合物信号范围内，N-乙酰半乳糖胺等碳水化合物成分减少；在1 230~1 300 cm^-1的酰胺Ⅲ带信号范围内蛋白质二级构象发生显著改变，β-折叠显著增加，α-螺旋减少；在2 800~3 100 cm^-1的脂质信号范围内，长链脂肪酸含量下降，支链脂肪酸含量上升。综上所述，玻璃化冷冻会改变猪卵母细胞蛋白质二级结构、脂肪酸组成及N-乙酰半乳糖胺等关键成分，导致蛋白质结构损伤、脂质组成改变以及碳水化合物代谢异常等多重冷冻损伤。

关键词: 猪, 卵母细胞, 拉曼光谱, 玻璃化冷冻

Abstract:

This study aimed to investigate molecular structural changes in porcine mature oocytes before and after vitrification using Raman spectroscopy. Ovaries were collected from pigs at a slaughterhouse, and cumulus-oocyte complexes were isolated in the laboratory. After in vitro maturation, fresh oocytes and vitrified oocytes were collected, respectively. Laser confocal Raman spectrometer was used to obtain the Raman spectrum information of fresh oocytes and vitrified oocytes. The molecular structure changes of oocytes before and after vitrification were analyzed after quality control of the data. The results indicated that, compared to fresh oocytes, vitrified oocytes exhibited reduced carbohydrate components such as N-acetylgalactosamine within the carbohydrate signal range of 1 020-1 140 cm^-1. In the amide Ⅲ band signal range (1 230-1 300 cm^-1), β-sheet content increased markedly while α-helix structures decreased. Within the lipid signal range (2 800-3 100 cm^-1), the content of long-chain fatty acids decreased, while the content of branched-chain fatty acids increased. In summary, vitrification alters the secondary protein structure, fatty acid composition and other key components like N-acetylgalactosamine in porcine oocytes. These changes result in multiple cryodamage effects, including protein structural damage, altered lipid composition, and abnormal carbohydrate metabolism.

Key words: porcine, oocyte, Raman spectrum, vitrification

中图分类号:

S828.3

陈思颍, 李伉, 孙雅雯, 冷璇, 王栋, 庞云渭. 基于拉曼光谱分析猪卵母细胞玻璃化冷冻前后变化[J]. 畜牧兽医学报, 2025, 56(10): 4998-5006.

CHEN Siying, LI Kang, SUN Yawen, LENG Xuan, WANG Dong, PANG Yunwei. Analysis of Changes of Porcine Oocytes before and after Vitrification Based on Raman Spectroscopy[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 4998-5006.

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峰/(cm^-1) Peak	核酸 DNA/RNA	蛋白质 Protein	脂质 Lipid	碳水化合物 Carbohydrate	其他成分 Others
944		C-C BK str
1 004		sym ring br Phe
1 097	sym str PO^2-	C-N str			sym str PO₄^3-
1 128		C-N str		C-O str
1 020~1 140		C-N str		C-O str, C-O-H br
1 262	T, A	Amide Ⅲ	═C-H ben
1 303		CH₃/CH₂ twi, ben	CH₂ twi, ben
1 340			C-H def
1 444		CH def	CH def	CH def
1 659		Amide Ⅰ	C═C str
1 747			C═O str^[25]
2 820~3 000		sym str CH₃/CH₂	sym str CH₃/CH₂

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