桑色素通过下调脂多糖刺激的原代牛乳腺上皮细胞中核因子-κB和丝裂原活化蛋白激酶(MAPK)信号通路来抑制炎性细胞因子的表达。

Morin suppresses inflammatory cytokine expression by downregulation of nuclear factor-κB and mitogen-activated protein kinase (MAPK) signaling pathways in lipopolysaccharide-stimulated primary bovine mammary epithelial cells.

作者信息

Wang Jingjing, Guo Changming, Wei Zhengkai, He Xuexiu, Kou Jinhua, Zhou Ershun, Yang Zhengtao, Fu Yunhe

机构信息

College of Veterinary Medicine, Jilin University, Jilin, Changchun 130062, P. R. China.

College of Veterinary Medicine, Jilin University, Jilin, Changchun 130062, P. R. China.

出版信息

J Dairy Sci. 2016 Apr;99(4):3016-3022. doi: 10.3168/jds.2015-10330. Epub 2016 Feb 3.

Abstract

Morin, a flavonoid isolated from Chinese herbs of the Moraceae family, has been reported to possess antiinflammatory activity. However, the effects of morin on mastitis have not been investigated. The present study was conducted to elucidate the antiinflammatory properties of morin on lipopolysaccharide (LPS)-stimulated primary bovine mammary epithelial cells (bMEC). The viability of bMEC was analyzed by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium] assay. Subsequently, bMEC were stimulated with LPS in the presence or absence of morin. Gene expression of proinflammatory cytokines was determined by quantitative real-time PCR (qRT-PCR). Nuclear factor-κB (NF-κB), inhibitory kappa B (IκBα) protein, extracellular signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK) were detected by Western blotting. The results showed that cell viability was not affected by morin. Moreover, morin inhibited the gene expression of tumor necrosis factor-α (TNF-α), IL-6, and IL-1β in LPS-stimulated bMEC in a dose-dependent manner. Western blot analysis showed that morin suppressed the phosphorylation of IκBα, NF-κB unit p65, ERK, p38, and JNK in LPS-stimulated bMEC. In conclusion, the protective effects of morin on LPS-induced inflammatory response in bMEC may be due to its ability to suppress NF-κB and mitogen-activated protein kinase (MAPK) signaling pathways. These findings suggest that morin may be used as antiinflammatory drug for mastitis.

摘要

桑色素是从桑科中草药中分离出的一种黄酮类化合物,据报道具有抗炎活性。然而,桑色素对乳腺炎的影响尚未得到研究。本研究旨在阐明桑色素对脂多糖(LPS)刺激的原代牛乳腺上皮细胞(bMEC)的抗炎特性。通过MTT [3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑] 法分析bMEC的活力。随后,在有或没有桑色素的情况下用LPS刺激bMEC。通过定量实时PCR(qRT-PCR)测定促炎细胞因子的基因表达。通过蛋白质印迹法检测核因子-κB(NF-κB)、抑制性κB(IκBα)蛋白、细胞外信号调节激酶(ERK)、p38和c-Jun氨基末端激酶(JNK)。结果表明,桑色素不影响细胞活力。此外,桑色素以剂量依赖性方式抑制LPS刺激的bMEC中肿瘤坏死因子-α(TNF-α)、IL-6和IL-1β的基因表达。蛋白质印迹分析表明,桑色素抑制LPS刺激的bMEC中IκBα、NF-κB亚基p65、ERK、p38和JNK的磷酸化。总之,桑色素对LPS诱导的bMEC炎症反应的保护作用可能归因于其抑制NF-κB和丝裂原活化蛋白激酶(MAPK)信号通路的能力。这些发现表明,桑色素可能用作乳腺炎的抗炎药物。

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