摘要
本试验旨在探究茶多酚是否激活沉默信息调节因子2相关酶3(SIRT3)缓解氧化应激引发的奶牛乳腺上皮细胞(BMECs)炎性反应。通过对SIRT3进行干扰并采用过氧化氢(H_(2)O_(2))损伤BMECs后进行茶多酚(TP)干预,明确TP激活SIRT3对氧化损伤BMECs炎性反应的作用。结果表明:干扰SIRT3后用H_(2)O_(2)处理导致BMECs线粒体功能障碍,极显著降低核呼吸因子1(NRF1)、过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)、线粒体转录因子A(TFAM)等线粒体因子的mRNA相对表达量(P<0.01),进而引起细胞发生氧化应激,极显著上调BMECs中氧化应激标志物丙二醛(MDA)含量和活性氧(ROS)荧光强度(P<0.01),导致抗氧化信号通路线粒体核呼吸因子2(NRF2)、血红素氧合酶-1(HO-1)的mRNA相对表达量极显著下降(P<0.01),进而极显著降低BMECs中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)含量(P<0.01),极显著提高炎性因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)的mRNA相对表达量(P<0.01),最终引起细胞凋亡因子B细胞淋巴瘤-2相关X蛋白(BAX)、半胱氨酸天冬氨酸蛋白酶3(CASP3)等因子mRNA相对表达量极显著升高(P<0.01),导致BMECs凋亡。加入TP后可逆转这一反应,通过提高抗氧化因子和线粒体因子的含量,抑制炎性因子和凋亡因子的表达,缓解由H_(2)O_(2)诱导BMECs产生的氧化应激和炎性反应。而当敲除SIRT3后这种抑制作用消失,线粒体功能障碍和氧化损伤加剧。综上所述,本研究揭示了TP通过激活SIRT3缓解H_(2)O_(2)诱导的BMECs线粒体功能障碍、氧化应激及炎性反应的作用与机制,为TP在奶牛生产中的推广应用提供了坚实的理论基础和技术支撑。
This experiment aimed to explore whether tea polyphenols could activate silent information regulator 2-related enzyme 3(SIRT3)to alleviate the inflammatory response in bovine mammary epithelial cells(BMECs)induced by oxidative stress.By interfering with SIRT3,damaging BMECs with hydrogen peroxide(H_(2)O_(2)),and then intervening with tea polyphenols(TP),the effect of TP activating SIRT3 on the inflammatory response of oxidatively damaged BMECs was clarified.The results showed that after interfering with SIRT3 and treating with H_(2)O_(2)lead to mitochondrial dysfunction,the mRNA relative expression levels of mitochondrial factors such as nuclear respiratory factor 1(NRF1),peroxisome proliferator-activated receptor-gamma coactivator-1α(PGC-1α)and mitochondrial transcription factor A(TFAM)were significantly decreased(P<0.01),which in turn caused oxidative stress in cells,the content of malondialdehyde(MDA),an oxidative stress marker in BMECs,and the fluorescence intensity of reactive oxygen species(ROS)were significantly increased(P<0.01),the mRNA relative expression levels of the antioxidant pathway nuclear respiratory factor 2(NRF2)and heme oxygenase-1(HO-1)were significantly decreased(P<0.01),the contents of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in BMECs were significantly decreased(P<0.01),the mRNA relative expression levels of inflammatory factors interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-8(IL-8)were significantly increased(P<0.01),finally,the mRNA relative expression levels of apoptosis factors such as B-cell lymphoma 2-associated X protein(BAX)and cysteine-containing aspartate-specific protease 3(CASP3)were significantly increased(P<0.01),leading to the apoptosis of BMECs.However,the addition of TP could reverse this reaction,by increasing the contents of antioxidant factors and mitochondrial factors,and inhibiting the expression of inflammatory factors and apoptosis factors,alleviated the oxidative stress and inflammatory response of BMECs induced by H_(2)O_(2).When SIRT3 was knocked out,this inhibitory effect disappeared,and mitochondrial dysfunction and oxidative damage were aggravated.In conclusion,this study reveals the effect and mechanism of TP in alleviating the mitochondrial dysfunction,oxidative stress and inflammatory response of BMECs induced by H_(2)O_(2)through activating SIRT3,providing a solid theoretical and technical support for the popularization and application of TP in dairy cow production.
作者
冀思同
马学虎
安彦昊
马敏
马燕芬
JI Sitong;MA Xuehu;AN Yanhao;MA Min;MA Yanfen(College of Animal Science and Technology,Ningxia University,Yinchuan 750021,China;Ningxia Ruminant Germplasm Research Technology Co.,Ltd.,Yinchuan 750009,China)
出处
《动物营养学报》
北大核心
2026年第1期682-692,共11页
CHINESE JOURNAL OF ANIMAL NUTRITION
基金
宁夏反刍动物营养科技创新团队项目(2024CXTD008)
银川市奶牛高效健康养殖科研创新团队项目(2023CXTD32)。
关键词
茶多酚
奶牛乳腺上皮细胞
氧化应激
炎性损伤
线粒体功能障碍
tea polyphenols
bovine mammary epithelial cells
oxidative stress
inflammatory damage
mitochondrial dysfunction
