松果体及其褪黑素对大鼠下丘脑CRH神经元的影响

发布时间：2018-01-20 08:21

本文关键词： 松果体褪黑素大鼠室旁核 CRH神经元正中隆起原位杂交免疫组织化学 ELISA　出处：《福建医科大学》2005年硕士论文　论文类型：学位论文

【摘要】：目的: 研究松果体及其褪黑素对大鼠下丘脑室旁核CRH 神经元形态和功能的影响以及补充褪黑素的作用,丰富松果体及褪黑素对下丘脑——垂体——肾上腺轴影响的形态及功能方面的研究,为进一步揭示松果体及褪黑素对免疫功能以及免疫相关性疾病如肿瘤、自身免疫性疾病等的影响机制提供理论依据。方法: ①运用CRHmRNA 原位杂交的方法来观察松果体摘除及补充外源性褪黑素后下丘脑室旁核CRHmRNA 阳性细胞的变化;②应用免疫组织化学染色的方法观察松果体摘除及补充外源性褪黑素后下丘脑室旁核CRH 阳性细胞和神经垂体正中隆起CRH 阳性染色的变化;③运用ELISA 法检测松果体摘除及补充褪黑素后血清CRH 浓度的变化。结果: ①松果体摘除后大鼠下丘脑室旁核CRH 原位杂交和免疫组化的结果与假手术对照组相比均显示阳性细胞数增多、阳性面积增加、平均灰度降低,提示松果体摘除后下丘脑CRH 神经元的表达增强;②松果体摘除大鼠补充褪黑素后,下丘脑CRH 原位杂交和免疫组化染色增强呈剂量依赖性恢复,不过不论是低剂量组还是高剂量组,也不论是术后4 周还是术后8 周,均不能恢复至正常;③松果体摘除后神经垂体正中隆起CRH 免疫组化结果显示阳性面积增加、平均灰度降低,提示下丘脑CRH 神经元释放CRH 增加;④ELISA 法检测血清CRH结果示各组血清CRH 浓度差异没有显著性,表明松果体摘除及补充褪黑素对血清CRH 水平没有显著影响。结论: 松果体摘除导致大鼠下丘脑室旁核CRH 神经元合成和释放CRH 增加,并通过垂体门脉系统激活整条HPA 轴,而补充外源性褪黑素后能在一定程度上逆转上述异常,表明松果体是神经——内分泌——免疫网络的结构和功能的重要稳定因素之一。
[Abstract]:Objective: to study the effects of pineal body and melatonin on the morphology and function of CRH neurons in hypothalamic paraventricular nucleus (PVN) of rats and the effects of melatonin supplementation. Studies on the morphology and function of pineal and melatonin on hypothalamus-pituitary-adrenal axis, in order to further reveal the immune function of pineal and melatonin and immune-related diseases such as tumors. The influence mechanism of autoimmune disease and so on provides the theoretical basis. Methods: 1the changes of CRHmRNA positive cells in hypothalamic paraventricular nucleus after pinealectomy and exogenous melatonin supplementation were observed by CRHmRNA in situ hybridization. 2Immunohistochemical staining was used to observe the changes of CRH positive cells and CRH positive staining in the paraventricular nucleus of hypothalamus after pinealectomy and supplementation of exogenous melatonin. 3 the changes of serum CRH levels after pinealectomy and melatonin supplementation were detected by ELISA method. Results: 1 the results of CRH in situ hybridization and immunohistochemistry in the paraventricular nucleus of hypothalamus after pinealectomy showed that the number of positive cells increased, the positive area increased and the average gray level decreased compared with the sham-operated control group. The results suggested that the expression of CRH neurons in hypothalamus was increased after pinealectomy. 2After the pinealectomy rats were supplemented with melatonin, the enhancement of CRH in situ hybridization and immunohistochemical staining in hypothalamus recovered in a dose-dependent manner, but both in low dose group and high dose group. No matter 4 weeks or 8 weeks after operation, they could not return to normal. 3After pinealectomy, the CRH immunohistochemical results of neurohypophyseal median eminence showed that the positive area increased and the average gray level decreased, suggesting that the CRH release from CRH neurons in the hypothalamus increased. The results of Elisa showed that there was no significant difference in serum CRH concentration among the three groups, indicating that pinealectomy and melatonin supplementation had no significant effect on serum CRH level. Conclusion: pinealectomy can increase the synthesis and release of CRH from CRH neurons in the paraventricular nucleus of hypothalamus and activate the whole HPA axis through the pituitary portal system. These abnormalities can be reversed to a certain extent by supplementation of exogenous melatonin, which indicates that the pineal gland is one of the important stable factors in the structure and function of neuro-endocrine immune network.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R33

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