姜黄素缓解热应激诱导的奶牛乳腺上皮细胞氧化应激与凋亡的作用研究

doi:10.11843/j.issn.0366-6964.2025.12.042

摘要/Abstract

摘要：

旨在评估姜黄素(curcumin, Cur)对热应激(heat stress, HS)诱导的奶牛乳腺上皮细胞氧化应激和凋亡的保护作用，并探讨其通过Nrf2信号通路调节相关基因表达的潜在机制。首先，通过不同浓度姜黄素(0、5、10、20、50 nmol·L^-1)处理, 筛选对细胞无毒性作用的浓度；其次，在热应激条件下，筛选细胞活力恢复最显著的浓度作为后续处理浓度。将奶牛乳腺上皮细胞分为对照组(NC)、姜黄素处理组(Cur)、热应激组(HS)和姜黄素处理的热应激组(HS+Cur), 采用免疫荧光、Western blot和RT-qPCR等方法检测氧化应激反应、凋亡情况及泌乳相关基因表达。结果表明，姜黄素在5和10 nmol·L^-1浓度下对奶牛乳腺上皮细胞活力无显著影响(P>0.05)，但在10 nmol·L^-1浓度下显著提高了热应激条件下的细胞活力(P < 0.001)。姜黄素通过上调线粒体膜电位、降低ROS和H₂O₂水平、显著提高抗氧化酶(SOD、GSH)活性，减少丙二醛(MDA)含量，显著抑制Bax/Bcl-2比值(P < 0.05)，从而减少细胞凋亡。Western blot结果表明，姜黄素通过降低Keap1的表达并上调Nrf2、HO-1(P < 0.01)和NQO1(P < 0.05)等下游抗氧化基因的表达。RT-qPCR结果显示，姜黄素显著恢复了热应激条件下泌乳相关基因(如CSN1S1、CSN1S2、CSN2等)的表达(P < 0.05)。综上，姜黄素通过调控Nrf2信号通路，缓解热应激对牛乳腺上皮细胞的氧化应激和凋亡损伤，恢复细胞活力并上调泌乳相关基因的表达，为姜黄素在缓解奶牛热应激中的应用提供了理论依据。

关键词: 姜黄素, 热应激, 奶牛乳腺上皮细胞

Abstract:

This study aims to evaluate the protective effects of curcumin (Cur) on oxidative stress and apoptosis induced by heat stress (HS) in bovine mammary epithelial cells and to explore its potential mechanism via the regulation of related gene expression through the Nrf2 signaling pathway. Different concentrations of curcumin (0, 5, 10, 20, 50 nmol·L^-1) were used to screen the concentration which doesn't have cytotoxic effect on the cells. Then the concentration that showed the most significant recovery in cell viability under heat stress was selected as the treatment concentration for subsequent experiment. Bovine mammary epithelial cells were divided into four groups: control group (NC), curcumin-treated group (Cur), heat stress group (HS), and curcumin-treated heat stress group (HS+Cur). And oxidative stress response, apoptosis, and lactation-related gene expression were assessed using immunofluorescence, Western blot, and RT-qPCR. The results showed that, curcumin at concentrations of 5 and 10 nmol·L^-1 did not significantly affect the viability of bovine mammary epithelial cells (P>0.05). However, 10 nmol·L^-1 curcumin significantly improved cell viability under heat stress conditions (P < 0.001). Curcumin increased mitochondrial membrane potential, reduced ROS and H₂O₂ levels, enhanced the activity of antioxidant enzymes (SOD and GSH), decreased malondialdehyde (MDA) levels, and significantly inhibited the Bax/Bcl-2 ratio (P < 0.05), thereby reducing apoptosis. Western blot results showed that curcumin reduced Keap1 expression while upregulating Nrf2, HO-1 (P < 0.01) and NQO1 (P < 0.05) downstream antioxidant gene expression. RT-qPCR results demonstrated that curcumin significantly restored the expression of lactation-related genes (CSN1S1, CSN1S2, CSN2) under heat stress conditions (P < 0.05).In conclusion, curcumin alleviates oxidative stress and apoptosis induced by heat stress in bovine mammary epithelial cells by modulating the Nrf2 signaling pathway. It restores cell viability and upregulates lactation-related gene expression, providing a theoretical basis for the application of curcumin in mitigating heat stress in dairy cows.

Key words: curcumin, heat stress, bovine mammary epithelial cells

中图分类号:

S816.7

伏庭书, 周宏达, 王苗, 陈梦炜, 柏小楠, 马保华, 彭莎. 姜黄素缓解热应激诱导的奶牛乳腺上皮细胞氧化应激与凋亡的作用研究[J]. 畜牧兽医学报, 2025, 56(12): 6411-6421.

FU Tingshu, ZHOU Hongda, WANG Miao, CHEN Mengwei, BAI Xiaonan, MA Baohua, PENG Sha. Curcumin Alleviates Oxidative Stress and Apoptosis Induced by Heat Stress in Bovine Mammary Epithelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6411-6421.

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基因 Gene	基因登录号 Accession No.	引物序列(5′→3′) Primers sequence (5′→3′)	产物大小/bp Product size
β-酪蛋白基因CSN2	NM_174524.2	5′-TGAGTCCCTCTTGCCATTCT-3′ 5′-GCTGGTCCAGCAGAGTACAT-3′	180
αS1-酪蛋白基因CSN1S1	NM_174528.2	5′-ACACAGGAGGACTCCAGGAA-3′ 5′-GTGGTGACATAGGCAGGAAG-3′	200
αS2-酪蛋白基因CSN1S2	NM_174529.2	5′-ATGACAGACCTCAGCACCAC-3′ 5′-AGTAGGCTGGCTTGGGTTTA-3′	180
κ-酪蛋白基因CSN3	NM_174527.2	5′-GCTGGGTACAGTATTATGAGGACAC-3′ 5′-GTTTGTGGAAGCGGTGTAGC-3′	150
泌乳蛋白基因BLG	NM_181024.2	5′-CTGGAAGAGTGAGGAGAACAG-3′ 5′-TCCCTGAGTTCTGCTTGAAC-3′	200
催乳素受体基因PRLR	NM_181024.2	5′-CGGCAACTTCAGACACACTG-3′ 5′-GCCAGCTTCTTTCGATGTCA-3′	190
泌乳素基因LALBA	NM_174539.3	5′-GGTGTCCAGTTCCTGTGCTA-3′ 5′-GCACCCAGTGGAAGTAAAGG-3′	180
β-actin基因ACTB	NM_001098478.1	5′-CCACGAGACCACCTTCAACT-3′ 5′-GTACAGGGACAGCACAGCCT-3′	180

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