焦虑模型大鼠下丘脑—垂体—肾上腺轴高反应性与皮质类固醇受体关系的研究

发布时间：2018-03-20 14:08

本文选题：焦虑模型　切入点：HPA轴　出处：《重庆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:研究在应激条件下焦虑模型大鼠下丘脑-垂体-肾上腺(hypothalamus-pituitary-adrenal,HPA)轴呈高反应性是与轴本身功能亢进还是与皮质类固醇受体改变相关。方法:将SD健康雄性大鼠随机分为两组,对照组(n=20)和模型组(n=26)。采用不可预知的情绪应激方法建立大鼠焦虑模型,对照组给予正常饲养。每周称量所有大鼠的体重,并在造模结束后,通过行为学实验检测造模是否成功。高架十字迷宫和旷场作为应激源刺激大鼠,测定两组大鼠血清中大鼠促肾上腺皮质激素(ACTH)和皮质酮(CORT)水平。用实时荧光定量PCR技术检测两组大鼠海马组织中盐皮质激素受体(mineralocorticoid steroid receptor,MR)和糖皮质激素受体(glucocorticoid steroid receptor,GR)m RNA水平及GR在下丘脑、垂体和肾上腺组织中的m RNA表达水平。蛋白免疫印迹(Western blot)检测两组大鼠海马组织内MR和GR蛋白水平以及GR在下丘脑、垂体和肾上腺组织中的蛋白表达水平。结果:(1)与对照组相比,焦虑模型大鼠体重增长率显著降低(p0.01)。(2)高架十字迷宫和旷场实验结果表明,与对照组相比,模型组大鼠进入开放臂的次数(OE)、开放臂停留的时间(OT)、进入开放臂的次数百分比(OE%)和进入开放臂的时间百分比(OT%)均显著降低(p0.01);模型组大鼠运动的总路程、中央活动路程百分比和中央活动时间百分比均显著降低(p0.01,p0.05)。(3)在基础条件下,两组大鼠分泌的促肾上腺皮质激素和皮质酮无明显差异(p0.05);而在应激条件下,模型组大鼠分泌的两种激素增加显著高于对照组(p0.01)。(4)与对照组相比,模型组MR m RNA在大鼠海马组织中表达无明显差异(p0.05);而模型组大鼠GR在海马、下丘脑、垂体和肾上腺组织中的m RNA表达水平均显著降低(p0.01,p0.05)。(5)与对照组比较,模型组MR蛋白表达在大鼠海马组织中无明显差异(p0.05);GR蛋白表达在大鼠海马、下丘脑和肾上腺组织中均显著降低(p0.01,p0.05),而在垂体中表达无差异(p0.05)。结论:在应激条件下,焦虑模型大鼠HPA轴呈高反应性与GR介导的轴负反馈调节作用改变相关。
[Abstract]:Objective: To study the anxiety under stress in rat model of hypothalamic pituitary adrenal axis (hypothalamus-pituitary-adrenal, HPA) showed high reactivity with the shaft itself or hyperfunction associated with corticosteroid receptor changes. Methods: healthy male SD rats were randomly divided into two groups, the control group (n=20) and model group (n=26) to establish a rat model of anxiety. The emotional stress method is unpredictable, the control group was given normal diet. All rats were weighed every week in weight, and after the end of the modeling by behavioral experiments testing the success of modeling. The elevated plus maze and open field as a source of stress stimulation in rats, the determination of the two groups of rats in the serum of rats with adrenocorticotropic hormone (ACTH) and corticosterone (CORT) levels. Detected by real-time fluorescence quantitative PCR in the hippocampus of rats in the two groups mineralocorticoid receptor (mineralocorticoid steroid, receptor, MR) and sugar Glucocorticoid receptor (glucocorticoid steroid, receptor, GR) m RNA level and GR expression level of M RNA in hypothalamus, pituitary and adrenal tissues. Western blot (Western blot) detection in the hippocampus of rats in the two groups of MR and GR protein level and GR expression level in hypothalamus, pituitary and adrenal tissue protein results: (1) compared with the control group, the anxiety model rats weight growth rate decreased significantly (P0.01). (2) the elevated plus maze and open field test results show that, compared with the control group, the number of rats in the open arm (OE), open arm time (OT). The percentage of times in the open arm (OE%) and in the open arm time percentage (OT%) were significantly decreased (P0.01); the total distance of rats in the model group movement, the percentage of the central distance percentage and central activity time were significantly decreased (P0.01, P0.05). (3) based on conditions Next, two groups of adrenocorticotropic hormone and corticosterone secretion in rats had no significant difference (P0.05); while under stress condition, two kinds of models of hormone secretion in rats group increased significantly higher than the control group (P0.01). (4) compared with the control group, model group, MR m RNA in rat hippocampal tissue there was no difference between the expressions (P0.05); while the rats in the model group GR in hippocampus, hypothalamus and pituitary, the expression level of M RNA in adrenal glands were significantly decreased (P0.01, P0.05). (5) compared with the control group, model group, the expression of MR protein in hippocampus of rats had no obvious difference (P0.05); the expression of GR protein in rat hippocampus, hypothalamus and adrenal tissues were significantly decreased (P0.01, P0.05), and no difference in pituitary expression (P0.05). Conclusion: under stress, anxiety model rat HPA Axis axis showed high reactivity and negative feedback regulation mediated by GR is related to the change of.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.72;R-332

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