摘要
目的采用逍遥散干预慢性应激损伤模型大鼠,观察其海马区神经PAS域蛋白2(neuronal PASdomain protein 2,Npas2)基因的表达及其DNA特定区域甲基化的修饰情况。方法将90只雄性Wistar大鼠随机分为空白对照组、慢性应激模型组和逍遥散治疗组,共3组,每组30只。造模采用Cart多相慢性应激大鼠模型并加以改进,逍遥散治疗组在造模同时每天给予逍遥散2 m L,连续造模21 d。采用RT-PCR一步法及免疫组织化学染色法测定各组大鼠海马区Npas2基因及其蛋白表达情况,基因甲基化检测采用SequenomMass ARRAY法。结果慢性应激损伤大鼠海马Npas2m RNA及其相应蛋白表达有明显变化,与空白对照组和逍遥散治疗组比较差异均有统计学意义,但其m RNA及相应蛋白表达变化趋势不一致,其基因某些特定区域的甲基化程度有升高的趋势(个别位点有统计学差异)。逍遥散治疗能够明显逆转慢性应激引起的Npas2m RNA及其相应蛋白的表达变化,相应区域DNA甲基化程度表现出下调趋势(个别位点有统计学差异)。结论 Npas2基因参与了慢性应激损伤过程,其表达变化可能与其基因某些特定位点的甲基化修饰变化有关,逍遥散能够逆转这种表达改变,可能是通过调节Npas2基因特定位点的甲基化程度发挥作用。
Objective In this study,Xiaoyao san was used to interfere with chronic stress injury model in rats,andthe expression of Npas2 gene and the methylation of DNA in the hippocampus were observed. Methods A total of90 male Wistar rats were randomly divided into three groups,control group,chronically stressed model group and Xiaoyao san treatment group. Improved Cart multi-phase chronically stressed rat model was used;rats of Xiaoyao san treatment group were fed with Xiaoyao san 2 m L each daily at the same time of modeling for 21 days. Theexpression of Npas2 m RNA and protein in hippocampus of rats were determined by RT-PCR andimmunohistochemical staining, and the Npas2 gene methylation was detected by the Sequenom Mass ARRAYmethod. Results The expression of Npas2 m RNA and its protein in hippocampus of rats with chronic stress injurywere significantly changed compared with the control group and the Xiaoyao san treatment group,but the expressiontrend of the m RNA and the protein were inconsistent. The degree of methylation of certain regions of its DNA has increased（the difference of individual sites was statistically significant）. The changes in the expression of Npas2 m RNA and its protein induced by chronic stress could be reversed significantly by treating with Xiaoyao san,andthe degree of DNA methylation in the corresponding region showed a downward trend（there were statisticallysignificant differences in individual sites）. Conclusion Npas2 gene is involved in the process of chronic stressinjury,and its expression may be related to the methylation modification of some specific sites on the gene,and thechanges of the expression of Npas2 gene can be reversed by treating with Xiaoyao san,that possibly be effectivethrough modulating the degree of methylation of specific sites on Npas2 gene.
作者
孙琪
郭维
江东
马会明
马榕
陈宏
SUN Qi1,2, GUO Wei1,2, JIANG Dong1, MA Huiming3, MA Rong1, CHEN Hong1,2(1. College of Traditional Chinese Medicine, Ningxia Medical University, Yinchuan 750004 Ningxia, China; 2. Key Laboratory of Hui Medicine, Ningxia Medical University, Yinchuan 750004 Ningxia, China; 3. Key Laboratory of Fertility Preservation, Ningxia Medical University, Yinchuan 750004 Ningxia, Chin)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2018年第4期421-427,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金(81260515)
