利用牛乳腺细胞和小鼠乳腺组织分析异绿原酸C通过NF-κB信号通路对乳腺炎症反应的抑制效应

doi:10.11843/j.issn.0366-6964.2023.09.031

摘要/Abstract

摘要： 旨在探究异绿原酸C(ICAC)对乳腺炎及NF-κB炎性通路的抑制作用。本研究以脂多糖(LPS)分别刺激牛乳腺上皮细胞(MAC-T)和小鼠乳腺组织建立体外和体内炎性模型,采用MTT方法筛选ICAC处理MAC-T细胞的适宜浓度,ELISA检测炎性因子(IL-1β、IL-6和TNF-α)和炎性介导因子(COX-2和iNOS)的表达水平,Western blot方法检测NF-κB信号通路关键蛋白(IκB和p65)的表达水平和磷酸化水平。结果发现:1)ICAC在20~100 mg·L^-1的浓度下对MAC-T细胞没有显著的生长抑制作用(P>0.05);2)1 mg·L^-1的LPS处理引起MAC-T细胞中IL-1β、IL-6的蛋白质表达量极显著高于对照组(P<0.01),ICAC处理(20、50、80 mg·L^-1)极显著下调了MAC-T细胞中IL-1β、IL-6、TNF-α、COX-2和iNOS的表达量(P<0.01);3)小鼠腹腔注射的ICAC(60、80、100 mg·kg^-1)均降低了LPS诱导的CD3的表达,小鼠乳腺组织T淋巴细胞浸润减少;4)ICAC显著降低了LPS诱导的MAC-T细胞和小鼠乳腺组织中p65和IκB的磷酸化水平(P<0.05),抑制了p65的核转移。上述结果说明,异绿原酸C通过NF-κB炎性通路抑制MAC-T细胞和小鼠乳腺组织的炎性反应,但是异绿原酸C在奶牛乳腺炎防治中的应用有待进一步研究。

关键词: 异绿原酸C, 抗炎作用, NF-κB, MAC-T, 小鼠乳腺

Abstract: The research aims to investigate the inhibitory effect of isochlorogenic acid C (ICAC) on mastitis and NF-κB inflammatory pathway. In this study, bovine mammary epithelial cells (MAC-T) and mouse mammary tissue stimulated with lipopolysaccharide (LPS) were used as in vitro and in vivo inflammatory model, respectively. MTT method was used to screen the optimal concentrations of ICAC for MAC-T cell treatment. The expression levels of inflammatory factors (IL-1β, IL-6 and TNF-α) and inflammatory mediators (COX-2 and iNOS) were detected using ELISA, whereas the expression and phosphorylation levels of key proteins (IκB and p65) in NF-κB signaling pathway were detected by Western blot. The results showed that:1) ICAC did not significantly inhibit the growth of MAC-T cells at the concentrations from 20 to 100 mg·L^-1 (P>0.05); 2) The expression levels of IL-1β and IL-6 protein in MAC-T cells treated with 1 mg·L^-1 LPS were significantly higher than those in untreated group (P<0.01), but ICAC (20, 50, 80 mg·L^-1) significantly down-regulated the expression of IL-1β, IL-6, TNF-α, COX-2 and iNOS in MAC-T cells (P<0.01); 3) Intraperitoneal inject ICAC(60、80、100 mg·kg^-1) reduced LPS-induced CD3 expression and T lymphocyte infiltration in mammary tissues of mice; 4) ICAC significantly reduced the phosphorylation of p65 and IκB in LPS-induced MAC-T cells and in mouse mammary tissue (P<0.05), and it inhibited the nuclear transfer of p65. These results indicate that ICAC inhibits the inflammatory response in MAC-T cells and mouse mammary tissue through NF-κB inflammatory pathway, but the role of ICAC in the prevention and treatment of bovine mastitis needs further investigation.

Key words: isochlorogenic acid C, anti-inflammatory, NF-κB, MAC-T, mouse mammary gland

中图分类号:

S858.23

陈喜宏, 路桂聪, 王浩磊, 苟少校, 玉永雄, 林涛, 蒋曹德. 利用牛乳腺细胞和小鼠乳腺组织分析异绿原酸C通过NF-κB信号通路对乳腺炎症反应的抑制效应[J]. 畜牧兽医学报, 2023, 54(9): 3931-3940.

CHEN Xihong, LU Guicong, WANG Haolei, GOU Shaoxiao, YU Yongxiong, LIN Tao, JIANG Caode. Isochlorogenic Acid C Inhibits Mammary Inflammatory Response through NF-κB Signaling Pathway Using Bovine Mammary Gland Cells and Mouse Mammary Gland Tissue[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3931-3940.

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