【摘要】： 老年人普遍存在記憶、認(rèn)知功能的下降。導(dǎo)致老年大腦記憶、認(rèn)知功能下降的機(jī)制是多方面的。其中,血管因素可能是造成老年大腦功能衰退的重要機(jī)制之一。大腦毛細(xì)血管是腦血管樹的最細(xì)小分支互相吻合而成的一個(gè)三維網(wǎng)狀結(jié)構(gòu),對(duì)調(diào)節(jié)大腦微環(huán)境、維持局部腦血流量起重要作用,毛細(xì)血管周圍微環(huán)境中神經(jīng)元的正常功能活動(dòng)、神經(jīng)沖動(dòng)的有效傳遞等均有賴于毛細(xì)血管完整的形態(tài)結(jié)構(gòu)。已有大量的定性研究深入細(xì)致的觀察了老年大腦不同部位毛細(xì)血管形態(tài)的變化,如毛細(xì)血管扭曲、球形改變及線樣改變等大體形態(tài)的變化,以及內(nèi)皮細(xì)胞萎縮,基膜增厚,足細(xì)胞退化等超微結(jié)構(gòu)改變。而關(guān)于大腦毛細(xì)血管的定量研究較少,且多限于大腦皮質(zhì)、海馬等部位。近年來,運(yùn)用新的體視學(xué)方法的研究發(fā)現(xiàn)正常老年大腦并不存在顯著性大腦皮質(zhì)神經(jīng)元死亡。那么,皮質(zhì)毛細(xì)血管是否發(fā)生老年性改變?盡管有過此方面的研究,但這些研究的結(jié)果差異較大,且均沒有貫徹現(xiàn)代體視學(xué)方法的原理。此外,運(yùn)用體視學(xué)方法的研究結(jié)果表明老年大腦白質(zhì)體積發(fā)生顯著性萎縮,白質(zhì)內(nèi)有髓神經(jīng)纖維的總長度顯著下降。此時(shí),白質(zhì)毛細(xì)血管發(fā)生了那些改變?國內(nèi)外尚未見報(bào)道。因此,基于對(duì)以往皮質(zhì)毛細(xì)血管老年性改變的研究不足之處的總結(jié)以及對(duì)白質(zhì)毛細(xì)血管老年性改變這一空白區(qū)域的補(bǔ)充,本實(shí)驗(yàn)以大鼠作為動(dòng)物模型,利用體視學(xué)定量研究方法和免疫組化技術(shù)對(duì)大腦皮質(zhì)、白質(zhì)內(nèi)毛細(xì)血管的老年性改變進(jìn)行了研究。 第一部分大鼠大腦皮質(zhì)毛細(xì)血管老年性改變的體視學(xué)研究 1材料與方法 1.1雌性Long-Evans大鼠,青年組(7月齡)10只,老年組(27月齡)10只。 1.2將大鼠麻醉灌注固定后,從每只大鼠隨機(jī)抽取一側(cè)大腦半球,制作2 mm厚的連續(xù)腦組織塊,從每個(gè)腦塊切取一張4μm厚的各向同性切片,進(jìn)行免疫組化染色。在100倍油鏡下從每張切片皮質(zhì)區(qū)域隨機(jī)選取3-5個(gè)視野拍照,每只動(dòng)物共拍攝30張照片。 1.3運(yùn)用體視學(xué)方法分別計(jì)算出大鼠大腦皮質(zhì)總體積、皮質(zhì)體積皺縮率、皮質(zhì)內(nèi)毛細(xì)血管長度密度和總長度、體積密度和總體積、表面積密度和總表面積。 2結(jié)果 2.1青年組、老年組雌性大鼠皮質(zhì)由于組織處理引起的體積皺縮率分別是53.9±7.3%及48.8±8.8%。青年組大鼠大腦組織處理前皮質(zhì)總體積為457.5±37.0 mm~3,老年組大鼠大腦組織處理前皮質(zhì)總體積為426.3±63.6 mm3,兩組之間無顯著性差異(p0.05)。 2.2青年組大鼠皮質(zhì)毛細(xì)血管的總長度為331.9±62.2 m,老年組大鼠皮質(zhì)毛細(xì)血管的總長度為281.6±61.5 m。兩組大鼠皮質(zhì)毛細(xì)血管的總長度無顯著性差異(p 0.05)。 2.3青年組大鼠皮質(zhì)毛細(xì)血管的總體積為6.7±1.2 mm3,老年組大鼠皮質(zhì)毛細(xì)血管的總體積為6.0±1.2 mm3。兩組大鼠皮質(zhì)毛細(xì)血管的總體積無顯著性差異(p 0.05)。 2.4青年組大鼠皮質(zhì)毛細(xì)血管的總表面積為46.5±9.8 cm2,老年組大鼠皮質(zhì)毛細(xì)血管的總表面積為41.7±9.4 cm2。兩組大鼠皮質(zhì)毛細(xì)血管的總表面積無顯著性差異(p 0.05)。 第二部分大鼠大腦白質(zhì)毛細(xì)血管老年性改變的體視學(xué)研究 1材料與方法 1.1雌性Long-Evans大鼠,青年組(7月齡)10只,老年組(27月齡)10只。 1.2將大鼠麻醉灌注固定后,從每只大鼠隨機(jī)抽取一側(cè)大腦半球,制作2 mm厚的連續(xù)腦組織塊,從每個(gè)腦塊切取一張4μm厚的各向同性切片,進(jìn)行免疫組化染色。在100倍油鏡下拍每張切片的整個(gè)白質(zhì)區(qū)域,拍得4-6個(gè)視野,每只動(dòng)物共拍攝30張照片。 1.3運(yùn)用體視學(xué)方法分別計(jì)算出大鼠大腦白質(zhì)總體積、白質(zhì)體積皺縮率、白質(zhì)內(nèi)毛細(xì)血管長度密度和總長度、體積密度和總體積、表面積密度和總表面積。 2結(jié)果 2.1青年組、老年組雌性大鼠白質(zhì)由于組織處理引起的體積皺縮率分別是46.4±8.8%及46.1±7.3%。青年組大鼠大腦組織處理前白質(zhì)總體積為91.7±12.1 mm~3,老年組大鼠大腦組織處理前白質(zhì)總體積為78.0±16.4 mm~3,老年組比青年組顯著降低了14.9%(p 0.05)。 2.2青年組大鼠白質(zhì)毛細(xì)血管的總長度為34.4±7.8 m,老年組大鼠白質(zhì)毛細(xì)血管的總長度為23.3±5.1 m。老年組比青年組顯著降低了32.2%(p 0.01)。 2.3青年組大鼠白質(zhì)毛細(xì)血管的總體積為0.81±0.13 mm~3,老年組大鼠白質(zhì)毛細(xì)血管的總體積為0.54±0.14 mm~3。老年組比青年組顯著降低了33.2%(p 0.01)。 2.4青年組大鼠白質(zhì)毛細(xì)血管的總表面積為5.6±1.0 cm2,老年組大鼠白質(zhì)毛細(xì)血管的總表面積為3.8±0.89 cm2。老年組比青年組顯著降低了31.7%(p 0.01)。 全文結(jié)論 1.本研究首次把新的體視學(xué)方法和免疫組化技術(shù)結(jié)合起來定量研究大鼠大腦皮質(zhì)毛細(xì)血管、白質(zhì)毛細(xì)血管的老年性改變。 2.大鼠大腦皮質(zhì)毛細(xì)血管總長度、總體積及總表面積不存在顯著的老年性改變。 3.老年大鼠白質(zhì)毛細(xì)血管總長度、總體積及總表面積顯著降低。 4.首次獲得了青年及老年雌性大鼠大腦皮質(zhì)毛細(xì)血管及白質(zhì)毛細(xì)血管的總長度、總體積及總表面積的基本數(shù)值,這些數(shù)值為今后運(yùn)用該動(dòng)物模型探討中樞神經(jīng)系統(tǒng)退化性疾病對(duì)皮質(zhì)、白質(zhì)內(nèi)毛細(xì)血管的影響提供了重要的參考數(shù)據(jù)。
[Abstract]:The prevalence of memory and cognitive decline in the elderly. The mechanism that leads to the memory of the old brain and the decline of cognitive function is multi-faceted. Among them, vascular factors may be one of the important mechanisms to cause brain function decline in old age. the brain capillary is a three-dimensional network structure which is matched with the smallest branches of the cerebral vascular tree and plays an important role in regulating the microenvironment of the brain, maintaining local cerebral blood flow, and normal functional activity of neurons in the microenvironment surrounding the capillary, The effective transmission of nerve impulse depends on the morphological structure of capillary vessel. The changes of capillary morphology in different parts of the brain, such as capillary distortion, spherical change and line-like change, were observed in a large number of qualitative researches, and the ultrastructural changes of endothelial cell atrophy, thickening of basement membrane and degeneration of foot cells were observed. However, the quantitative study on the capillaries of the brain is less and is not limited to cerebral cortex, hippocampus and so on. In recent years, it has been found that there is no significant cerebral cortex neuron death in the normal old brain using the new method of stereology. So, do cortical capillaries have age-related changes? Although there are studies in this field, the results of these studies are quite different and do not carry out the principle of modern stereology. In addition, the study of the method of stereology showed that the white matter volume in the elderly decreased significantly, and the total length of myelinated nerve fibers in white matter decreased significantly. At this point, white matter capillaries have changed? No reports have been reported at home and abroad. Therefore, on the basis of the summary of the deficiency of the research on the senile changes of the capillary capillary in the past and the supplement to the white matter capillary age, this experiment takes rat as animal model, uses the stereological quantitative research method and the immunohistochemical technique to the cerebral cortex, Age-related changes of capillary in white matter are studied. Changes of capillaries in cerebral cortex of rat cerebral cortex in the first part visual study Materials and Methods 1. 1 female Long-Evans rat, young group (7 months old) 10, old Ten rabbits (27 months old) were randomly extracted one side of cerebral hemisphere from each rat, and a 2 mm thick continuous brain tissue mass was randomly extracted from each rat, and a 4. m u.m thickness was cut from each brain block. Isotropic slices were stained with immunohistochemical staining. 3-5 fields of vision were randomly selected from each slice cortex region under 100-fold oil mirror The total volume of cerebral cortex, volume shrinkage of cortex, length density and total length of capillary in cortex were calculated by stereological method. bulk density degree and total volume, surface area density, and total surface area. Results 2. 1 young group, the cortex of female rats in the old group was caused by tissue treatment. The volume shrinkage rate was 53. 9% 7. 3% and 48. 8% 8. 8% respectively. The total volume of cortex before treatment of cerebral tissue in young rats was 457. 5 mm37. 0 mm ~ 3, and the total volume of cortex before brain tissue treatment in the aged rats was 426. The total length of cortex capillary in the aged rats was 331. 9/ 62. 2 m, and the total length of capillary capillary in the aged rats was 281. 6 mm61. There was no significant difference in the total length of cortex capillaries in the two groups (p 0.05). The total volume of capillary capillary in the aged rats was 6. 0. 2 mm3. There was no significant difference in the total volume of cortex capillaries in the two groups (p 0.05). The total surface area of the cortical capillaries in the young rats was 4. 6.5 The total surface area of the cortex capillaries in the aged rats was 4. 8 cm2 and 9. 8 cm2. 1. 7 B9. 4 cm2. Total surface of the cortical capillaries of the two sets of rats No significant difference in product Stereological study of age-related changes in cerebral white matter capillary in the second part of rats (p 0.05) Materials and Methods 1. 1 female Long-Evans rats, 10 young adults (7 months old), 10 elderly (27 months old) group (27 months old). Rats were randomly drawn from one side of the brain hemisphere to create a 2 mm thick continuous brain tissue block from each brain block with a 4. m u.m thick each. the whole white matter area of each slice is photographed under 100 times of oil mirror, 4-6 fields of vision are taken, 30 photos are taken for every animal, The method The total volume of white matter, the volume shrinkage of white matter, the length density and total length, volume density and total volume, surface area density and total volume of white matter in the white matter were calculated. Results 2. Results The volume shrinkage of white matter caused by tissue treatment was 46. 4% 8. 8% and 46. 1% 7. 3%, respectively. The total white matter volume was 91% in young rats. The total volume of white matter before brain tissue treatment in aged group was 78. 0 65.16. 4 mm ~ 3. The old group was significantly lower than that of young group by 14.9% (p 0.05). The total length of white matter capillary was 34. 4, 7. 8 m, and the total length of white matter capillary in the old group was 23. 3/ 5. 1m. The old age group was significantly lower than that of the young group.% (p 0.01). 2. 3 youth group white matter capillary The total volume of blood vessels was 0. 81, 0. 13 mm ~ 3, and the total volume of white matter capillary in the aged rats was 0. 54% 0. 14 mm ~ 3. The old age group was significantly higher than the young group. a decrease of 30.2% (p 0.01). 2. 4 young people Group rat white The total surface area of the capillary was 5. 6 cm 1. 0 cm2, and the total surface area of white matter capillary in the old group was 3. 8 mm0. 89 cm2. 31 7% (p 0.01) were significantly lower than that in the young group (p 0.01). For the first time in this study, the new method of stereology and immunohistochemical technique were combined in quantitative study. Age-related changes in capillary and white matter capillaries in the cerebral cortex of rats. The total length, total volume and total surface area of capillaries in the cerebral cortex of rats did not have significant age-related changes. 3. The total length of white matter capillary in the aged rats the total volume and the total surface area were significantly reduced.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R322

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