【摘要】：目的：研究在雙側(cè)卵巢切除并補充雌激素(ovariectomized，estrogen primed，OEP)的小鼠側(cè)腦室注射瘦素(leptin)后，神經(jīng)肽Y(Neuropeptide Y，NPY)Y1受體免疫陽性細(xì)胞，Y1受體(NPY Y1R)蛋白以及血清中卵泡刺激素(follicle stimulating hormone，F(xiàn)SH)和黃體生成素(luteinizing hormone，LH)的變化，，探討leptin調(diào)控生殖的可能機理，為leptin的臨床用藥提供一定的實驗依據(jù)。 方法：選用清潔級成年雌性C57BL/6J小鼠60只，制備OEP模型：在無菌條件下行雙側(cè)卵巢切除術(shù)，術(shù)后第11天開始每天皮下注射苯甲雌二醇(Estradiol Benzoate，E_2)2μg/只，連續(xù)注射10天。OEP模型完成后將小鼠隨機分成實驗組和對照組，定位側(cè)腦室，實驗組注射含4μg leptin的生理鹽水2μl，對照組注射同體積生理鹽水，每組又分別按注射后1.0h，1.5h，2.0h，2.5h分為4個組。 1．將上述2.5h實驗組和對照組小鼠經(jīng)左心室穿刺進(jìn)入主動脈，刺破右心耳，用生理鹽水沖洗，4％多聚甲醛快速灌注，以視交叉和乳頭體后緣的冠狀切面為界取腦組織，常規(guī)冰凍切片，免疫組織化學(xué)方法觀察NPY Y1R免疫陽性細(xì)胞在小鼠下丘腦的分布和變化。 2．將各時間段實驗組和對照組小鼠摘除眼球取血，離心后取上層血清，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.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R33

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