去卵巢及雌激素替代治疗后家兔边缘系统NOS的活性及nNOS阳性神经元的形态结构与分布变化
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摘要
为了探讨雌激素、孕激素对去卵巢家兔边缘系统NOS的活性及对nNOS阳性神经元形态结构分布的影响,本试验将家兔切除卵巢,作为妇女更年期后卵巢功能衰竭、雌激素分泌严重不足时的良好模型,然后通过人为注射雌激素及雌激素和孕激素,观察去卵巢和注射雌激素及雌激素和孕激素后家兔血清中NO的浓度、边缘系统NOS活性及nNOS阳性神经元的形态结构分布变化来研究雌激素保护中枢神经系统、预防老年痴呆和记忆力减退等神经系疾病的机理,为神经生理学和病理学研究提供形态学依据。
利用SABC免疫组织化学法,在光学显微镜下对各组家兔海马CA1、CA2、CA3区及齿状回、下丘脑室旁核、下丘脑外侧区、视上核以及额叶神经型一氧化氮合酶(nNOS)阳性神经元的形态结构及分布进行观测。结果表明:假手术对照组家兔海马、下丘脑、额叶内有大量的nNOS阳性神经元分布;去卵巢后nNOS阳性神经元形态分布变化有区域差异,额叶和海马各区(除CA2外)nNOS阳性神经元表达明显减弱,下丘脑各神经核nNOS阳性神经元的表达明显增强,即数量增多,染色加深;雌激素替代治疗组与假手术组之间无显著差异,额叶和海马各区(除CA2外)nNOS阳性神经元表达明显增强,下丘脑各神经核nNOS阳性神经元的表达明显减弱,即数量减少,染色变浅。
利用硝酸还原酶法和生物化学检测技术,采用NO、NOS试剂盒分别检测去卵巢和注射雌激素及雌激素和孕激素后家兔血清中NO的浓度的变化及大脑边缘系统NOS的活性变化。结果表明:去卵巢三个月后家兔血清NO水平和大脑边缘系统NOS活性降低;单纯补充雌激素和联合使用雌激素、孕激素时,血清NO水平和大脑边缘系统NOS活性均升高;单纯补充雌激素血清NO水平和大脑边缘系统NOS的活性高于联合使用雌激素、孕激素时血清NO水平和大脑边缘系统NOS活性,但差异不显著。
To study the effect of estradiol benzoate and progesterone on the activity of NOS and distribution and structure of nNOS positive neurons in Rabbit limbic system, female rabbits were spayed as models of women who are in perimenopausal period in this experiment. The comparation of NO solution and the changes of NOS activity and expression of nNOS in rabbit limbic system were observed after estrogen replacement therapy and combined hormone replacement. All this research work will provide a basis for the study of Menopausal Syndrome and nervous system disease, especially the study of the physiology of nervous pathology.
The first periment studied the change of nNOS-positive neurons in the sections of the hippocampus and in PVN、LHA、SON of hypothalamus and frontal lobe under light microscope following ovariectomy and estrogen replenish. Results: There were comprehensive nNOS-positive staining cell groups in all regions of hippocampus and hypothalamus and frontal lobe. After ovariectomy, the changes of nNOS positive neurons in subregion were not equal: the obvious to die down of the nNOS expression in frontal lobe and in all regions of hippocampus but CA2, while the obvious to build up of the nNOS expression in the each nerve pit of hypothalamus, namely a increasing amount and a deeper dyeing. There was no significant difference between estradiol-replacement therapy and the control group. The obvious to build up of the nNOS expression in frontal lobe and in all regions of hippocampus but CA2, and the obvious to weaken of the nNOS expression in each nerve pit of hypothalamus, namely a decreasing amount and a lighter dyeing compared with the ovariectomy group.
The second experiment studied NO levels and the activity of NOS in rabbit limbic system after ovariectomy and estrogen replacemen therapy and combined hormone replacement by the method of nitric acid reductase and metabolic chemistral method. Results: there was a significant decrease of NO levels and the activity of NOS compared with control group after ovariectomy. NO level and the activity of NOS increased after using estrogen only and group of combined hormone replacement. NO level and the activity of NOS after using estrogen only were hingher than the group of combined hormone replacement,but there was no significant difference.
利用SABC免疫组织化学法,在光学显微镜下对各组家兔海马CA1、CA2、CA3区及齿状回、下丘脑室旁核、下丘脑外侧区、视上核以及额叶神经型一氧化氮合酶(nNOS)阳性神经元的形态结构及分布进行观测。结果表明:假手术对照组家兔海马、下丘脑、额叶内有大量的nNOS阳性神经元分布;去卵巢后nNOS阳性神经元形态分布变化有区域差异,额叶和海马各区(除CA2外)nNOS阳性神经元表达明显减弱,下丘脑各神经核nNOS阳性神经元的表达明显增强,即数量增多,染色加深;雌激素替代治疗组与假手术组之间无显著差异,额叶和海马各区(除CA2外)nNOS阳性神经元表达明显增强,下丘脑各神经核nNOS阳性神经元的表达明显减弱,即数量减少,染色变浅。
利用硝酸还原酶法和生物化学检测技术,采用NO、NOS试剂盒分别检测去卵巢和注射雌激素及雌激素和孕激素后家兔血清中NO的浓度的变化及大脑边缘系统NOS的活性变化。结果表明:去卵巢三个月后家兔血清NO水平和大脑边缘系统NOS活性降低;单纯补充雌激素和联合使用雌激素、孕激素时,血清NO水平和大脑边缘系统NOS活性均升高;单纯补充雌激素血清NO水平和大脑边缘系统NOS的活性高于联合使用雌激素、孕激素时血清NO水平和大脑边缘系统NOS活性,但差异不显著。
To study the effect of estradiol benzoate and progesterone on the activity of NOS and distribution and structure of nNOS positive neurons in Rabbit limbic system, female rabbits were spayed as models of women who are in perimenopausal period in this experiment. The comparation of NO solution and the changes of NOS activity and expression of nNOS in rabbit limbic system were observed after estrogen replacement therapy and combined hormone replacement. All this research work will provide a basis for the study of Menopausal Syndrome and nervous system disease, especially the study of the physiology of nervous pathology.
The first periment studied the change of nNOS-positive neurons in the sections of the hippocampus and in PVN、LHA、SON of hypothalamus and frontal lobe under light microscope following ovariectomy and estrogen replenish. Results: There were comprehensive nNOS-positive staining cell groups in all regions of hippocampus and hypothalamus and frontal lobe. After ovariectomy, the changes of nNOS positive neurons in subregion were not equal: the obvious to die down of the nNOS expression in frontal lobe and in all regions of hippocampus but CA2, while the obvious to build up of the nNOS expression in the each nerve pit of hypothalamus, namely a increasing amount and a deeper dyeing. There was no significant difference between estradiol-replacement therapy and the control group. The obvious to build up of the nNOS expression in frontal lobe and in all regions of hippocampus but CA2, and the obvious to weaken of the nNOS expression in each nerve pit of hypothalamus, namely a decreasing amount and a lighter dyeing compared with the ovariectomy group.
The second experiment studied NO levels and the activity of NOS in rabbit limbic system after ovariectomy and estrogen replacemen therapy and combined hormone replacement by the method of nitric acid reductase and metabolic chemistral method. Results: there was a significant decrease of NO levels and the activity of NOS compared with control group after ovariectomy. NO level and the activity of NOS increased after using estrogen only and group of combined hormone replacement. NO level and the activity of NOS after using estrogen only were hingher than the group of combined hormone replacement,but there was no significant difference.
引文
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包新民,舒斯石.大鼠脑立体定位图谱[M].北京:人民卫生出版社蔡溱,力夫编译.一氧化氮供体药物[J].国外医学药学分册, 2001, 8(4): 237-240
蔡薇薇,友群,激素与一氧化氮系统关系研究[M].中国药房, 2007,18 (23): 1824-1834
陈卫民,刘叔文.可释放一氧化氮化合物的研究进展与临床前景[J].解放军药学学报, 2000,16(5): 63-65
陈一勇.素对去势雌性大鼠海马结构内NOS阳性神经元的影响[D].山西医科大学硕士论文
杜业亮,郑培敏,熊克仁.大鼠下丘脑外侧区一氧化氮合酶阳性神经元的缺血后变化[J].皖南医学院学报,1998, 17(1): 4-6
方芳等.激素替代治疗后脑组织一氧化氮水平变化的试验研究2003, 1(18): 27-29
龚再菊,绝经后妇女骨密度变化与血清激素水平关系的探讨[J].现代康复, 2001,5(3): 86
皇甫平.雌激素对去势雌性大鼠下丘脑一氧化氮合酶阳性神经元分布的影响[D].山西医科大学硕士学位论文
李倬.一氧化氮的生物学意义及应用前景[J].西北民族学院学报, 2001, 2(3): 39-43
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