【作者】 吴连仲；

【导师】 韩卉； 姚国刚；

【作者基本信息】 安徽医科大学 ， 人体解剖与组织胚胎学， 2003， 硕士

【摘要】 目的 ① 观察高脂血症(hyperlipidemia)大鼠下丘脑神经肽Y(neuropeptide Y，NPY)免疫阳性神经元的形态学变化；② 测量高脂血症大鼠下丘脑NPY免疫阳性神经元的光密度(optical density，OD)、数量和截面积在高脂血症形成过程中的动态变化；③ 探讨高脂血症与下丘脑NPY免疫阳性神经元的关系，揭示下丘脑NPY免疫阳性神经元对脂质代谢的调节作用。方法 ① 将实验动物分为四组，对照组喂饲普通饮食，实验组分别喂饲高脂饮食4周，8周和12周，建立高脂血症动物模型；② 利用酶法测定大鼠的血清总胆固醇(total cholesterol，TC)和甘油三酯(triglyceride，TG)浓度；③ 神经元Nissl染色观察；④ 透射电镜观察；⑤ 免疫组化ABC染色观察；⑥ 计算机图像分析。结果 ① 血清平均TC，高脂饮食4周组为(1.941±0.493)mmol／L、8周组为(1.929±0.519)mmol／L，4周、8周组比对照组(1.295±0.398)mmol／L显著升高(P＜0.01)，12周组(1.35±0.210)mmol／L降至近于对照组水平(P＞0.05)。② 高脂饮食4周组血清平均TG浓度为(0.294±0.195)mmol／L，8周组为(0.239±0.126)mmol／L，4、8周组TG比对照组(0.508±0.216)mmol／L显著降低(P＜0.01)，12周TG(0.952±0.190)mmol／L显著升高(与各组比，P＜0.01)。③ 在下丘脑弓状核、室旁核、腹内侧核和背内侧核，高脂饮食8周、12周组神经元内有较多的Nissl颗粒，对照组和4周组神经元内的颗粒相对较少。各组细胞形态未见明显差别。④ 高脂饮食4周组大鼠下丘脑弓状核、室旁核、腹内侧核及背内侧核神经元内线粒体扩张，线粒体嵴紊乱或断裂，粗面内质网、滑面内质网及高尔基复合体呈囊样膨胀，溶酶体、游离核糖体较对照组增多。⑤ 高脂饮食4周、8周组下丘脑弓状核NPY免疫阳性神经元平均光密度(mean optical density，MOD)和数量显著低于对照组(P＜0.01)；12周组MOD为(0.2346±0.0316)，数量为(36.8±2.6)个，比对照组增高；NPY免疫阳性神经元截面积8周组为(89±2)μm~2、12周组为(86±3)μm~2，两者比对照组(120±4)μm~2显著安徽医科大学硕士论文吴连仲减小(P<0.01)。⑥4周组室旁核NPY免疫阳性神经元的MOD为(0 .1 265士0.0337)，数量为(9.3士2.6)个，显著低于对照组、12周组的MOD及细胞数(P<0.01)，4周组MOD与8周组(0.1 886士0.0302)比差异显著(P<0.01);12周组MOD(0 24巧士0.0208)比对照组增高;NPY免疫阳性神经元的截面积8周为(55士3)林mZ，12周为(50士2)协mZ，8、 xZ周显著小于对照组(106士2)脚2 (P<0.01)。⑦对照组、4周、8周组腹内侧核NPY免疫阳性神经元MOD无差异，12周的MOD(0.2327士0.0206)比4周(0.1901士0.0302)、8周(0.1992士0.0281)组显著升高(尸<0.01);4周、8周组免疫阳性神经元的平均数量分别为(5.1士1.3)个和(4.2士1 .6)个，比对照组(9.6士2.4)个、12周组显著减少;8周、12周组NPY免疫阳性神经元的截面积分别为(65土5)林时和(60土4)耐，比对照组(87士4)林扩显著减小。⑧4周组背内侧核NPY免疫阳性神经元MoD为(0.1766士0.0321)，比对照组(0.2185士0.0201)、12周组(0.2416士0.0286)显著降低(P<0.01);12周组MOD比8周组显著增高;8周组神经元数为(6.5士1 .2)个，少于对照组(8.6士1 .7)个和12周组(1 1 .5土1 .6)个咖<0.05)。8周、12周组NPY免疫阳性神经元的截面积分别为(75士3)。衬和(67士l)耐，显著小于对照组(92士5)脚2(尸<0.01)。结论①血清TC和TG呈时相性变化，总胆固醇浓度先升后降，甘油三酷浓度先降后升。②高脂血症导致下丘脑神经元的超微结构改变。③高脂血症导致下丘脑NPY免疫阳性神经元的光密度、数量和细胞的截面积发生改变。④下丘脑NPY免疫阳性神经元的平均光密度与血清总胆固醇浓度呈负相关趋势，与血清甘油三酷呈正相关趋势。⑤下丘脑NPY免疫阳性神经元参与脂质代谢的调节，对维持血脂稳定可能具有调节作用。更多还原

【Abstract】 Objectives (1)To observe the morphologic changes of the NPY immunoreactive neurons in hypothalamus of hyperlipidemia rats. (2)To estimate the changes of the NPY-immunoreactive neurons in hypothalamus during the high-fat diet .(3)To approach the effection of hyperlipidemia on hypothalamic NPY-immunoreactive neurons and to reveal the function of hypothalamic NPY-immunoreactive neurons in lipid metabolism. Methods (1)The animals were divided into four groups. The control group was feeded normal diet. The test groups were feeded high-fat diet for 4 weeks, 8 weeks and 12 weeks respectively to establish hyperlipidemia model. (2)Mensurating the concentration of serum total cholesterol(TC) and triglyceride(TG) by enzymatic method . (3)The Nissl-staining method. (4) Observing with transmission electron microscope. (5) Immunohistochemical ABC method was used. (6) The Computer Image Analyser was applied to. Results (1) The mean serum TC : In 4W and 8W groups the results were (1.941 0.493)mmol/L and (1.929 0.519)mmol/L respectively. The the mean serum TC of 4W or 8W group, was higher than that of the control group (1.295 0.398)mmol/L(P<0.01). The TC of 12W group was (1.35 0.210)mmol/L .descending near to the level of the control group. (2) The mean serum TG : In 4W and 8W groups, the results were (0.294 0.195)mmol/L and (0.239 0.126)mmol/L in turn, lower than control group (0.508 0.216)mmol/L (P<0.01). In 12W group, the mean serum TG was (0.952 0.190)mmol/L ,higher than that of other groups (P<0.01). (3) In ARC, PVN, VMN, DMN, there were more Nissl granules in 8W and12W groups than that in control and 4W groups . There was no significantly morphologic difference between each group.(4) In 4W group, The neurons showed a fat appearance. Mitochondria tumefacted and its bar was ruptured. Lysosomes and ribosome increased. Rough and smooth endoplasmic reticulum and Golgi complexes expanded. (5) In 4W and 8W groups, the mean optical density(MOD) and the number of NPY neurons in ARC were far lower than those of control group(P < 0.01); In 12W group, the MOD and the number of NPY neurons were (0.2346 0.0316) and (36.8 2.6) respectively, higher than those of control group (P< 0.05) ; In 8W group, the section area of neuron were (89.2 2) m2 . the section area of 12W group was (86 3) m2 , smaller than (120 4) m2 that of control group (P < 0.01). (6) The NPY neurons in PVN: In 4W group, the MOD and the number of NPY neurons were (0.1265 0.0337) and (9.3 2.6) respectively. The MOD and the neuron number of 4W group were much lower than that of control or 12W groups(p < 0.01 ),and the MOD of 4W group was different from 8W group markedly. In 12W group, the MOD of NPY neurons was (0.2415 0.0208), higher than that of control group(0.2016 0.0122), In 8W and 12W groups, the section area of neurons was much smaller than that of control group(106 4) um2 (P < 0.01). (7) There was no significant difference between the MOD of the NPY immunoreactive neurons in VMN of 4W and 8W and control groups, but the MOD of 12W group was (0.2327 0.0206),much higher than the MOD of 4W or 8W groups. The number of neurons in 8W group was (4.2 1.6),less than the number of control or 12W groups (p< 0.05) , there was no difference between 4W and 8W and 12W and control groups. In 8W and 12W groups, the section area of the neurons was much smaller than that of control group(. (8) The MOD of NPY immunoreactive neurons in DMN of 4W group was (0.1766 0.0302),far lower than that of control or 12W group (P < 0.01) ; The MOD of 12W group was(0.2327 0.0206) ,much higher than that of 4W or 8W groups. The neuron number of 8W group was (6.5 1.2),less than that of control or 12W group (p < 0.05) ; The section area of neurons of 8W or 12W groups was much smaller than that of control group(92 5) m2 (P< 0.01). Conclusions (1) The mean serum TC and TG changedwith the prolonging of the high-fat diet. The mean serum TC was higher at first and then lower, while the TG was lower at first and then higher. (2) There were ultrastructure changes of hypothalamic neurons. (3) Hyperlipidemia更多还原