瘦素对OEP小鼠下丘脑神经肽Y受体表达的影响
发布时间：2018-08-17 08:53
【摘要】：目的：研究在双侧卵巢切除并补充雌激素(ovariectomized，estrogen primed，OEP)的小鼠侧脑室注射瘦素(leptin)后，神经肽Y(Neuropeptide Y，NPY)Y1受体免疫阳性细胞，Y1受体(NPY Y1R)蛋白以及血清中卵泡刺激素(follicle stimulating hormone，FSH)和黄体生成素(luteinizing hormone，LH)的变化，，探讨leptin调控生殖的可能机理，为leptin的临床用药提供一定的实验依据。 方法：选用清洁级成年雌性C57BL/6J小鼠60只，制备OEP模型：在无菌条件下行双侧卵巢切除术，术后第11天开始每天皮下注射苯甲雌二醇(Estradiol Benzoate，E_2)2μg/只，连续注射10天。OEP模型完成后将小鼠随机分成实验组和对照组，定位侧脑室，实验组注射含4μg leptin的生理盐水2μl，对照组注射同体积生理盐水，每组又分别按注射后1.0h，1.5h，2.0h，2.5h分为4个组。 1．将上述2.5h实验组和对照组小鼠经左心室穿刺进入主动脉，刺破右心耳，用生理盐水冲洗，4％多聚甲醛快速灌注，以视交叉和乳头体后缘的冠状切面为界取脑组织，常规冰冻切片，免疫组织化学方法观察NPY Y1R免疫阳性细胞在小鼠下丘脑的分布和变化。 2．将各时间段实验组和对照组小鼠摘除眼球取血，离心后取上层血清，用放射免疫法(Radio-immunity analysis，
[Abstract]:Objective: to study the effects of leptin (leptin) on bilateral ovariectomized mice supplemented with estrogen primed (leptin). The changes of NPY Y1R protein and serum follicle stimulating hormone (follicle stimulating) and luteinizing hormone LH (LH) in immunoreactive cells of neuropeptide Y (Neuropeptide Y1 receptor were studied to explore the possible mechanism of leptin regulating reproduction and to provide some experimental evidences for the clinical use of leptin. Methods: 60 clean adult female C57BL/6J mice were used to establish OEP model. Bilateral ovariectomy was performed under aseptic conditions. Estradiol Benzoate E2 was injected subcutaneously every day on the 11th day after operation. After 10 days of continuous injection, mice were randomly divided into two groups: the experimental group and the control group. The rats in the experimental group were injected with 2 渭 L saline containing 4 渭 g leptin and the control group was injected with the same volume of saline. Each group was divided into four groups according to 1.0 h, 1.5 h, 2.0 h and 2.5 h after injection. 1. The mice of the experimental group and the control group were punctured into the aorta by the puncture of left ventricle, and the right atrial appendage was punctured. 4% paraformaldehyde was rapidly infused with normal saline. The coronal section of the optic chiasma and the posterior edge of the papillary body was used as the boundary to take the brain tissue, and the routine frozen section was used. Immunohistochemical method was used to observe the distribution and changes of NPY Y1R immunoreactive cells in the hypothalamus of mice. 2. Mice extirpate their eyeballs to get blood, After centrifugation, the upper serum was collected and Radio-immunity analysis was used.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R33
本文编号：2187105
[Abstract]:Objective: to study the effects of leptin (leptin) on bilateral ovariectomized mice supplemented with estrogen primed (leptin). The changes of NPY Y1R protein and serum follicle stimulating hormone (follicle stimulating) and luteinizing hormone LH (LH) in immunoreactive cells of neuropeptide Y (Neuropeptide Y1 receptor were studied to explore the possible mechanism of leptin regulating reproduction and to provide some experimental evidences for the clinical use of leptin. Methods: 60 clean adult female C57BL/6J mice were used to establish OEP model. Bilateral ovariectomy was performed under aseptic conditions. Estradiol Benzoate E2 was injected subcutaneously every day on the 11th day after operation. After 10 days of continuous injection, mice were randomly divided into two groups: the experimental group and the control group. The rats in the experimental group were injected with 2 渭 L saline containing 4 渭 g leptin and the control group was injected with the same volume of saline. Each group was divided into four groups according to 1.0 h, 1.5 h, 2.0 h and 2.5 h after injection. 1. The mice of the experimental group and the control group were punctured into the aorta by the puncture of left ventricle, and the right atrial appendage was punctured. 4% paraformaldehyde was rapidly infused with normal saline. The coronal section of the optic chiasma and the posterior edge of the papillary body was used as the boundary to take the brain tissue, and the routine frozen section was used. Immunohistochemical method was used to observe the distribution and changes of NPY Y1R immunoreactive cells in the hypothalamus of mice. 2. Mice extirpate their eyeballs to get blood, After centrifugation, the upper serum was collected and Radio-immunity analysis was used.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R33
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