LPS-induces IL-6 and IL-8 gene expression in bovine endometrial cells “through DNA methylation”
LPS-induces IL-6 and IL-8 gene expression in bovine endometrial cells “through DNA methylation”
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LPS 通过 DNA 甲基化诱导牛子宫内膜细胞中 IL-6 和 IL-8 基因表达
DOI：
10.1016/j.gene.2018.07.074
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发表时间：
2018
期刊：
影响因子：
3.5
通讯作者：
Wenfa Lu
中科院分区：
文献类型：
--
作者：
Jun Wang;Xiaoxiao Yan;Lucky T. Nesengani;He Ding;Lianyu Yang;Wenfa Lu
奶牛子宫感染造成巨大的经济损失。在牛子宫内膜细胞中，脂多糖（LPS）刺激的白细胞介素6 （IL-6）和白细胞介素8 (IL-8) mRNA的增加对炎症反应至关重要；然而，监管机制仍不清楚。在此，我们研究了DNA甲基化在lps诱导牛子宫内膜细胞IL-6和IL-8 mRNA表达中的作用。在LPS刺激后，用5-Aza-2 ' -脱氧胞嘧啶（5Aza）抑制DNA甲基化，评估IL-6和IL-8 mRNA的表达。采用实时荧光定量PCR （qRT-PCR）和亚硫酸盐测序PCR （BSP）分析LPS处理24 h后DNA甲基转移酶（DNMT1、DNMT3A和DNMT3B）、甲基cpg结合蛋白2 （MeCP2）的表达以及IL-6和IL-8区域的DNA甲基化。抑制DNA甲基化显著增强lps诱导的IL-6和IL-8 mRNA表达。24 h后，LPS增加了IL-6和IL-8 mRNA的表达，降低了IL-6启动子（- 366和- 660）和IL-8启动子（- 120和- 48）上特定CpG位点的甲基化水平。此外，LPS处理24 h显著增加DNMT1、DNMT3A、DNMT3B和MeCP2 mRNA的表达。结果表明，LPS处理牛子宫内膜细胞可诱导IL-6和IL-8启动子甲基化调控IL-6和IL-8 mRNA的表达。
Uterine infection in dairy cows causes great economic loss. In bovine endometrial cells, lipopolysaccharide (LPS)-stimulated increase in interleukin 6 (IL-6) and interleukin 8 (IL-8) mRNA is crucial for the inflammatory response; however, the regulatory mechanisms remain unclear. Here, we investigated the role of DNA methylation in IL-6 and IL-8 mRNA expression following LPS-induction in bovine endometrial cells. IL-6 and IL-8 mRNA expression was evaluated under DNA methylation inhibition using 5-Aza-2′-deoxycytodine (5Aza) following LPS stimulation. Expression of DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B), methyl CpG-binding protein 2 (MeCP2) and DNA methylation at IL-6 and IL-8 regions, were analyzed using quantitative real-time PCR (qRT-PCR) and bisulfite sequencing PCR (BSP) following 24 h of LPS treatment. Inhibition of DNA methylation significantly enhanced LPS-induced IL-6 and IL-8 mRNA expression. LPS increased IL-6 and IL-8 mRNA expression, and decreased methylation levels of specific CpG sites at the IL-6 promoter (at −366 and −660) and the IL-8 promoter (at −120 and −48) after 24 h. Furthermore, LPS treatment for 24 h significantly increased DNMT1, DNMT3A, DNMT3B, and MeCP2 mRNA expression. Our results indicate that treating bovine endometrial cells with LPS induces the expression of IL-6 and IL-8 mRNA regulated by IL-6 and IL-8 promoter methylation.