老年大鼠松果体结构特点及其生长抑素表达的形态学观察

发布时间：2018-01-31 18:52

本文关键词： 老年松果体生长抑素形态学大鼠　出处：《河北医科大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的:哺乳动物的松果体,是脑内重要的神经内分泌器官,可分泌多种单胺类和多肽类激素。随着年龄的增长,松果体出现结构变化和功能下降。本研究对老年大鼠松果体标本,进行光镜和电镜观察,揭示老年动物松果体细微和超微结构特点,以及生长抑素表达的变化,为深入研究松果体结构和功能的关系,提供形态学依据。方法: 1实验动物与取材选用健康老年(24月龄)SD大鼠15只,成年(3月龄)大鼠4只,雌雄不拘,由河北医科大学实验动物中心提供。于腹腔注射6%水合氯醛(0.5ml/100g)麻醉下,经升主动脉插管快速灌流0.9%温生理盐水,再用4℃的3%多聚甲醛和1%戊二醛混合固定液灌注约2-3小时,断头取脑后切取松果体浅部,置于3%多聚甲醛和1%戊二醛混合溶液(4℃)中浸泡固定48小时,用于光镜和电镜样品的制备。 2光镜样品制备与观察将醛类固定液固定的完整松果体标本,经梯度乙醇脱水,二甲苯透明,石蜡包埋,组织切片,行常规HE染色,进行光镜观察,用于显示松果体细微结构。 3免疫组织化学染色与观察石蜡包埋的松果体组织切片,行生长抑素免疫组织化学染色,光镜观察生长抑素及其受体,在松果体组织内各类细胞的表达。 4透射电镜样品制备与观察醛类固定的松果体标本,行常规透射电镜样品制备,超薄组织切片,进行透射电镜观察;以揭示老年大鼠松果体的超微结构特点。结果: 1石蜡包埋组织切片HE染色光镜观察显示: 松果体由实质和间质所组成。松果体实质主要由松果体细胞和少量胶质细胞构成;间质为疏松的结缔组织。青年组大鼠松果体细胞数目较多,排列紧密,胞核圆形,核仁明显。老年组大鼠松果体细胞数目减低,排列疏松,胞质内出现脂滴;并伴有明显的结缔组织增生。 2石蜡切片免疫组织化学染色光镜观察显示; 在青年组大鼠松果体组织切片,生长抑素免疫反应阳性细胞,以及阳性反应产物,主要位于松果体细胞的胞质;在松果体实质内还存在免疫反应阳性纤维。在老年组动物松果体组织切片,生长抑素免疫反应阳性细胞与对照组相似,阳性细胞为松果体细胞;免疫反应阳性产物不仅出现在胞质,在胞核内也有表达。 3超薄组织切片透射电镜观察显示; 老年大鼠松果体有松果体囊包被,主细胞松果体细胞形态不规则,有两个或多个突起,细胞间存在大量细胞间小管,小管相互之间,以及小管与松果体囊、血管周隙和结缔组织间隙相互连通;松果体细胞的胞核形态不规则,可见胞核分叶,核膜凹陷形成皱褶,异染色质明显增多,甚至呈现染色质边集现象,有些出现细胞凋亡样改变;有些细胞的部分线粒体肿胀变形,嵴溶解消失;常见少数细胞的内质网及高尔基复合体出现囊腔扩张,次级溶酶体、多泡小体增多,甚至出现大量的脂褐素颗粒沉积。大鼠松果体的毛细血管为有孔型毛细血管,血管内皮外有明显的基膜,基膜外是宽阔的结缔组织间隙,周细胞也比较常见。结论: 1大鼠松果体细胞为生长抑素免疫反应阳性细胞,老年大鼠松果体细胞对生长抑素的表达增强,表现在胞质和胞核同有表达。 2老年大鼠松果体呈现明显的细微和超微结构变化,主要表现在松果体细胞的减少,结缔组织间质成分的增多;松果体细胞内外脂质物质的沉积,以及脂褐素颗粒或斑块的形成。 3松果体组织结构,尤其是松果体细胞的形态学变化,可能与老年动物松果体功能的进行性减退有关。
[Abstract]:Objective: the pineal gland of mammals, is an important neuroendocrine organ in the brain, can secrete a variety of monoamine and peptide hormones. With the increase of age, the pineal gland appeared structural changes and functional decline. The study on the elderly rats were observed by light microscopy and electron microscopy, reveal the pineal gland and small animal the ultrastructural features and the changes of expression of somatostatin, for the study of the structure and function of the pineal gland, to provide morphological evidence.
Method:
1 experimental animals and materials
Healthy elderly (24 month old) 15 SD rats, adult (3 month old) 4 rats, male and female, provided by experimental animal center of Hebei Medical University. By intraperitoneal injection of 6% chloral hydrate (0.5ml/100g) under anesthesia by ascending aorta cannulation rapid perfusion 0.9% warm saline with 4 DEG C, 3% paraformaldehyde and 1% glutaraldehyde mixed stationary liquid perfusion for about 2-3 hours, decapitated behind cut pineal shallow, placed in 3% paraformaldehyde and 1% glutaraldehyde solution (4 DEG C) for fixed 48 hours, for light and electron microscopy sample preparation.
Preparation and observation of 2 light mirror samples
Samples of complete pineal gland fixed by aldehyde fixative were dehydrated by gradient ethanol, transparent with xylene and paraffin embedded. Tissue sections were stained with conventional HE. Light microscopy was used to observe the fine structure of pineal gland.
3 immunohistochemical staining and observation
Paraffin embedded pineal tissue slices were stained with somatostatin and immunohistochemistry. The expression of somatostatin and its receptors in pineal tissues was observed under light microscope.
Preparation and observation of 4 transmission electron microscope samples
The samples of pineal gland fixed by aldehydes were prepared by routine transmission electron microscopy, ultrathin sections and transmission electron microscope.
Result:
1 paraffin embedded tissue sections were observed by HE staining light microscopy.
The pineal gland is composed of parenchyma and stroma. The pineal parenchyma consists of pineal gland cells and a few glial cells; interstitial loose connective tissue. Many young rats pineal cells were arranged closely, round nuclei, prominent nucleoli. The aged rat pineal cell number decreased, loosely arranged and in the cytoplasm of lipid droplets; and with obvious hyperplasia of connective tissue.
2 paraffin sections were observed by immunohistochemical staining.
In the young rats body tissue, somatostatin immunoreactive cells, and positive reaction products, mainly located in the cytoplasm of the pineal gland cells in the pineal parenchyma; there are immunoreactive fibers in the aged group. Animal pineal tissue sections, somatostatin immunoreactive cells were similar to the control group. Positive cells in the pineal gland cells; immunoreactive not only appeared in the cytoplasm and in the nucleus is also expressed.
3 ultrathin tissue sections were observed by transmission electron microscopy.
The pineal body in aged rat pineal capsule were the main cells of the pineal gland cells were irregular, with two or more processes, there are a large number of cells between intercellular canaliculus, tubular and tubular and between the pineal capsule, and perivascular connective tissue space communicated with each other; the gland cells the nuclear morphology was irregular, lobulated nucleus visible, the formation of wrinkles nuclear membrane depression, heterochromatin increased significantly, or even a chromatin phenomenon, some cells appeared apoptosis like changes; some part of mitochondrial swelling deformation of cell, cristae disappeared; the endoplasmic reticulum and Golgi complex common few cells had cystic dilatation, secondary lysosomes, multivesicular bodies increased, and even the emergence of large deposits of lipofuscin granules. The rat pineal gland capillaries were fenestrated capillary, obvious vascular endothelial basement membrane, membrane is wide between the connective tissue The pericytes are also common in the gap.
Conclusion:
The pineal cells of 1 rats were somatostatin immunoreactive cells, and the expression of somatostatin in pineal cells of aged rats was enhanced, showing the same expression in cytoplasm and nuclei.
Show fine structure and ultrastructure changes of 2 senile rat pineal gland, mainly to reduce pineal cells, increase of substance between the connective tissue deposition; pineal extracellular lipid substances, as well as the formation of lipofuscin granules or plaque.
3 the histological structure of pineal body, especially the morphological changes of pineal cells, may be related to the progressive degeneration of the function of the pineal body in the old animals.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R322.8

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