甜菜碱缓解油酸诱导鸡胚原代肝细胞脂代谢紊乱及氧化应激的作用机制

doi:10.11843/j.issn.0366-6964.2025.09.050

摘要/Abstract

摘要：

旨在探究甜菜碱对鸡胚原代肝细胞(chicken embryo liver cells，CEL)氧化应激的调控作用。采用油酸诱导鸡胚原代肝细胞脂沉积和氧化应激的体外模型，通过添加甜菜碱探究其对鸡胚原代肝细胞氧化应激的调控机理。以鸡胚原代肝细胞为研究对象，分为对照组、油酸模型组和油酸模型+甜菜碱组(甜菜碱浓度为4、6、8、10 mmol·L^-1)，每组6个重复，处理24 h后用CCK-8法检测肝细胞增殖活力，并对细胞进行油红O染色，检测细胞内活性氧(reactive oxygen species, ROS)水平，同时收集细胞沉淀测定抗氧化相关指标。结果显示，4~10 mmol·L^-1甜菜碱对鸡胚原代肝细胞的增殖活力无显著影响(P>0.05)。油红O结果显示，油酸处理组肝细胞出现大量红色脂滴且总胆固醇(total cholesterol, TC)和甘油三脂(triglyceride, TG)含量均高于对照组(P＜0.05)；而与油酸组相比，4 mmol·L^-1甜菜碱组肝细胞内红色脂滴数量减少，同时TG含量显著降低(P＜0.05)。此外，与对照组相比，油酸组肝细胞丙二醛(malondialdehyde, MDA)和ROS水平显著升高(P＜0.05)，总抗氧化能力(total antioxidant capacity, T-AOC)有降低的趋势(P=0.083)；而添加4 mmol·L^-1甜菜碱后，鸡胚原代肝细胞的T-AOC水平高于油酸组，且MDA含量与油酸组相比显著降低(P＜0.05)。RT-PCR结果显示，油酸组抗氧化相关基因Nrf2、HO-1和NQO1基因的表达量显著低于对照组(P＜0.05)；与油酸组相比，添加4 mol·L^-1甜菜碱能显著升高Nrf2、HO-1、NQO1、CAT、SOD-1和GPX-1的mRNA丰度(P＜0.05)。本研究表明，4 mmol·L^-1甜菜碱对油酸诱导的鸡胚原代肝细胞的脂沉积和氧化应激有缓解作用，为甜菜碱在畜禽行业的应用提供理论依据。

关键词: 鸡胚原代肝细胞, 甜菜碱, 氧化应激, 脂沉积

Abstract:

The aim of this study was to investigate the regulatory effect of betaine on oxidative stress in chicken embryo liver cells (CEL). The study was carried out to explore the potential mechanism of betaine on oxidative stress and lipid metabolism in CEL with oxidative stress model induced by oleic acid. Cells were divided into control (Con) group, oleic acid (OA) group and OA+ betaine (OA+BT) groups (the betaine concentrations were 4, 6, 8 and 10 mmol·L^-1, respectively) according to different treatments, with 6 replicates in each group. After 24 h treatment, cell proliferation activity was detected by CCK-8 method. Oil red O staining was used to analyze the histopathological changes of the cells, and the intracellular reactive oxygen species (ROS) level was measured, and then cell pellets were collected to determine the antioxidant related indicators. The results showed that 4-10 mmol·L^-1 betaine had no significant effect on the proliferation activity of CEL cells (P>0.05). Oil red O results showed that a large number of red lipid droplets appeared in CEL cells in the OA group, and the contents of total cholesterol (TC) and triglyceride (TG) were higher than those in the Con group (P < 0.05). However, compared with the OA group, red lipid droplets in CEL cells were decreased in the 4 mmol·L^-1 OA+BT group, and the TG content was significantly decreased (P < 0.05). In addition, when compared with the Con group, the levels of malondialdehyde (MDA) and ROS were significantly increased (P < 0.05), and total antioxidant capacity (T-AOC) tended to decrease in the OA group (P=0.083). After adding 4 mmol·L^-1 betaine, the T-AOC level was higher than that in OA group, and the MDA content was significantly lower than that of OA group (P < 0.05). RT-PCR results showed that the expression levels of antioxidant related genes including Nrf2, HO-1 and NQO1 in the OA group were significantly lower than those in the Con group (P < 0.05). Compared with the OA group, the addition of 4 mmol·L^-1 betaine significantly increased the mRNA abundance of Nrf2, HO-1, NQO1, CAT, SOD-1 and GPX-1 (P < 0.05). The present study showed that 4 mmol·L^-1 betaine alleviated oleic acid-induced lipid deposition and oxidative stress in CEL cells, which will provide a theoretical basis for the application of betaine in the livestock and poultry industry.

Key words: chicken embryo liver cells, betaine, oxidative stress, lipoidosis

中图分类号:

S831.5

王超慧, 刘筱影, 杨小军, 刘艳利. 甜菜碱缓解油酸诱导鸡胚原代肝细胞脂代谢紊乱及氧化应激的作用机制[J]. 畜牧兽医学报, 2025, 56(9): 4741-4749.

WANG Chaohui, LIU Xiaoying, YANG Xiaojun, LIU Yanli. The Mechanism of Betaine in Alleviating Abnormal Lipid Metabolism and Oxidative Stress Induced by Oleic Acid in Chicken Embryo Liver Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4741-4749.

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基因 Gene	登录号 Accession number	引物序列(5′→3′) Primer sequences,	产物大小/bp Product size
β-actin	L_08165	F: ATTGTCCACCGCAAATGCTTC	113
		R: AAATAAAGCCATGCCAATCTCGTC
Nrf2	XM_046921130.1	F: AGCCACTTTATTCTTGCCTCT	117
		R: AAAATCATCAATCTCCCTGTCG
HO-1	NM_205344.2	F: ACACAACGCTGAAAGCATGT	163
		R: GATGAAGTACAGGGACGCCG
NQO1	NM_001277620.1	F: CGAGTGCTTTGTCTACGAGATGGAG	102
		R: AGGTCAGCCGCTTCAATCTTCTTC
CAT	NM_001031215.2	F: TCAGGAGATGTGCAGCGTTT	109
		R: TCTTACACAGCCTTTGGCGT
SOD-1	NM_205064.2	F: GGCAATGTGACTGCAAAGGG	133
		R: CCCCTCTACCCAGGTCATCA
GPX-1	NM_001277853.3	F: AGTACATCATCTGGTCGCCG	137
		R: CTCGATGTCGTCCTGCAGTT

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