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目的:研究REM期睡眠剥夺对成年大鼠不同脑区包括海马、腹内侧前额叶皮质、下丘脑和中缝核群的组蛋白H3K9和H3K4三甲基化水平的影响。方法:将成年SD大鼠随机分为正常对照组(con)、睡眠剥夺1 d组(SD1 d)、睡眠剥夺3 d组(SD3 d)、睡眠剥夺6 d组(SD6 d)。采用改良式多平台水环境法建立REM期睡眠剥夺模型,分别于睡眠剥夺后1、3、6 d断头取脑,用免疫荧光染色与Western Blot检测大鼠海马、腹内侧前额叶皮质、下丘脑和中缝核群H3K9和H3K4三甲基化水平的变化,最终结果进行统计学分析。结果:Western Blot结果显示:(1)海马、腹内侧前额叶皮质三个SD组H3K9三甲基化水平均低于control组(P<0.05),H3K4三甲基化水平均高于control组(P<0.05);(2)下丘脑三个SD组H3K9三甲基化水平均低于control组(P<0.01),但SD1 d组H3K4三甲基化水平高于control组(P<0.05),SD3 d、SD6 d组H3K4三甲基化水平低于control组(P<0.05);(3)中缝核群三个SD组H3K9和H3K4三甲基化水平均高于control组(P<0.05)。对海马与下丘脑免疫荧光验证H3K9三甲基化结果与Western Blot结果趋势一致,且说明细胞无减少。结论:睡眠剥夺可能与海马、腹内侧前额叶皮质、下丘脑和中缝核群的组蛋白H3K9和H3K4三甲基化水平密切相关。
AIM: To investigate the effects of REM sleep deprivation on histone H3K9 and H3K4 trimethylation levels in different brain regions including hippocampus, ventral medial prefrontal cortex, hypothalamus and meso-nucleus in adult rats. Methods: Adult SD rats were randomly divided into normal control group (SD), sleep deprivation for 1 d (SD1 d), sleep deprivation for 3 d (SD3 d) and sleep deprivation for 6 d (SD6 d). The REM sleep deprivation model was established by modified multi-platform water environment method. The brain was decapitated at 1, 3 and 6 days after sleep deprivation, and the hippocampus and ventral medial prefrontal cortex were detected by immunofluorescence and Western Blot. Changes in the levels of trimethylation of H3K9 and H3K4 in thalamus and meso-nucleus, and the final results were analyzed statistically. Results: The results of Western Blot showed that: (1) The levels of H3K9 trimethylation in hippocampus and ventral medial prefrontal cortex were lower than those in control group (P <0.05), and the levels of H3K4 trimethylation were higher than those in control group P <0.05). (3) The levels of H3K9 trimethylation in the hypothalamus three SD groups were lower than those in control group (P <0.01), but the levels of H3K4 trimethylation in SD1 d group were higher than those in control group (P <0.05) (P <0.05); (3) The levels of H3K9 and H3K4 trimethylation in three SD groups in SD group were significantly higher than those in control group (P <0.05) ). Immunofluorescence on hippocampus and hypothalamus showed that H3K9 trimethylation was consistent with Western Blot results and showed no reduction in cells. Conclusion: Sleep deprivation may be closely related to histone H3K9 and H3K4 trimethylation levels in hippocampus, ventral medial prefrontal cortex, hypothalamus and meso-nucleus.