小鼠大脑亚硝酸盐暴露与DNA甲基化和组蛋白去乙酰化

doi:10.16098/j.issn.0529-1356.2017.06.002

摘要/Abstract

摘要：

目的观察慢性亚硝酸盐暴露对小鼠大脑皮质炎症损伤的影响及其探讨DNA甲基化和组蛋白去乙酰化等相关机制。方法选取8周龄健康雄性C57BL/6J小鼠，随机分为对照组（生理盐水）、低剂量亚硝酸盐组（3g/L）和高剂量亚硝酸盐组(6g/L)，建立亚硝酸盐暴露模型，收集各组小鼠大脑皮质，利用免疫荧光染色法和Western blotting法分析大脑皮质炎症损伤，组蛋白去乙酰化酶和DNA甲基化相关酶的表达情况。结果慢性亚硝酸盐暴露后小鼠大脑皮质炎症损伤因子环氧化酶2（COX2）、白细胞介素-1β（IL-1β）、离子钙接头蛋白分子1（Iba1）、c-Fos、IL-6表达量明显多于对照组（P<0.01），同时高剂量暴露组DNA甲基化相关酶5-甲基胞嘧啶（5-mC）、DNA甲基转移酶1(DNMT1)、DNMT3a、和组蛋白去乙酰化酶1（HDAC1）表达明显低于对照组（P<0.01）且都呈亚硝酸盐剂量依赖性。结论亚硝酸盐暴露可通过促进细胞免疫炎症对小鼠大脑皮质造成损伤,并且DNA甲基化和组蛋白去乙酰化可能参与了慢性亚硝酸盐暴露过程中的应答过程及其调控机制。

关键词: 亚硝酸盐暴露, 炎症反应, DNA甲基化, 组蛋白去乙酰化, 免疫印迹法, 小鼠

Abstract:

Objective To investigate the inflammatory damage in the mouse neocortex after nitrite exposure, and to understand the regulation mechanism of DNA methylation and histone deacetylation. Methods C57BL/6 J adult male mice were used to establish the models of prenatal nitrite exposure, wherefore control group (physiological saline), low dose nitrite exposure group (3 g/L) and high dose nitrite exposure group (6 g/L) were included. Immunocytochemistry and Western blotting were carried out to analyze the inflammatory damage, histone deacetylation and DNA methylation in the neocortex. Results The expressions of cyclooxygenase 2(COX2), interleukin-1β(IL-1β), ionized calcium binding adapor molecule-1(Iba1), c-Fos and IL-6 were significantly more than those in the control group (P<0.01) after chronic nitrite exposure. The expression of 5-methylcytosine(5-mC), DNA methyctrasferace 1(DNMT1), DNA DNMT3a and histone deacetylase 1(HDAC1) in treatment groups were significantly lower than that in the control group (P<0.01) with dose dependency. Conclusion Nitrite exposure can induce cell immune inflammatory damage in the neocortex of mice, and DNA methylation and histone deacetylation may be involved in regulation of immune inflammatory reaction after nitrite exposure.

Key words: Nitrite exposure, Inflammatory reaction, DNA methylation, Histone deacetylation, Western blotting, Mouse

刘俊张丽丽石贞玉邓锦波. 小鼠大脑亚硝酸盐暴露与DNA甲基化和组蛋白去乙酰化[J]. 解剖学报, 2017, 48(6): 642-650.

LIU Jun ZHANG Li-li SHI Zhen-yu DENG Jin-bo. Nitrite exposure and DNA methylation and histone deacetylation in mouse brain[J]. Acta Anatomica Sinica, 2017, 48(6): 642-650.

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