长期跑步训练对大鼠大脑白质老年性改变的作用及机制探讨
发布时间:2018-05-27 01:40
本文选题:大脑白质及白质内有髓神经纤维 + 少突胶质细胞 ; 参考:《重庆医科大学》2012年博士论文
【摘要】:第一部分长期跑步对大鼠大脑白质内有髓神经纤维和少突胶质细胞老年改变的体视学研究 目的探讨从中年开始为期14个月的中强度跑步训练对大脑白质及其内有髓神经纤维和少突胶质细胞老年改变的作用,以期认识长期跑步锻炼保护大脑白质的形态学基础及其机制。 方法在跑步锻炼前(14月龄)、跑步4个月(18月龄)和跑步锻炼结束(28月龄)均随机挑选雌、雄性SD大鼠各10只进行水迷宫测试,行为学测试后进行形态学定量研究。随机挑选跑28月步锻炼组、对照组28月龄和对照组18月龄每组雌、雄性大鼠各5只,每只大鼠随机抽取一侧大脑半球,被抽取的每侧大脑半球随机抽取6块大小为1.5mm×1.5mm×1.0mm白质块制成60nm超薄切片,通过透射电子显微镜获得6-8张放大6000倍照片。余下的一侧大脑半球每隔1mm切取1张50μm冠状冰冻切片,标记切片中少突胶质细胞。运用体视学方法得到大鼠髓鞘老年改变的比例、大鼠大脑白质总体积、有髓神经纤维总长度、白质内有髓神经纤维总体积和少突胶质细胞数量。 结果1.跑步训练4个月的雌、雄性大鼠的潜伏期较跑步训练前显著缩短(p<0.05),跑步训练14个月的雌、雄性大鼠的潜伏期与跑步训练前比较没有显著性差异(p>0.05);长期锻炼后的潜伏期和对照组28月龄的潜伏期没有显著性差异(p>0.05)。2.28月龄跑步训练组、对照组18月龄和28月龄雌性大鼠大脑白质发生退行性改变的神经纤维比例分别为21.2±11.04%、35.8±9.78%和49.2±2.56%;28月龄跑步锻炼组、对照组18月龄和28月龄雄性大鼠大脑白质发生退行性改变的神经纤维比例分别为24.9±9.62%、50.80±3.64%和44.1±10.30%。与对照组大鼠比较,雌、雄性跑步训练组大脑白质发生退行性改变的有髓神经纤维比例均显著性降低(p<0.05)。3.28月龄雌、雄性跑步训练组大鼠大脑白质体积分别较对照组28月龄雌、雄性大鼠大56.5%(p<0.01)和43.3%(p<0.01)。28月龄跑步训练组雌性大鼠白质体积显著性大于照组18月龄雌性大鼠的白质体积(p<0.01),28月龄跑步训练组和对照组18月龄雄性大鼠白质体积没有显著性差异(p0.05)。4.跑步组雌、雄性大鼠大脑白质内有髓神经纤维总长度分别比对照组28月龄雌、雄性大鼠大脑白质内有髓神经纤维总长度长45.48%(p<0.01)和34.83%(p<0.01)。跑步组雌性大鼠比对照组18月龄雌性大鼠的白质有髓神经纤维总长度长48.20%(p<0.01);跑步组雄性大鼠比对照组18月龄雄性大鼠的白质有髓神经纤维总长度长23.52%(p0.05)。5.跑步组雌、雄性大鼠大脑白质内有髓神经纤维总体积较对照组28月龄雌、雄性大鼠分别大55.98%和46.84%(p<0.01);跑步组雌性大鼠比对照组18月龄雌性大鼠的白质内有髓神经纤维总体积大44.07%(p<0.01),跑步组雄性大鼠比对照组18月龄雄性大鼠的白质内有髓神经纤维总体积大20.57%(p0.05)。6.跑步组雌、雄性大鼠大脑白质内CNPase~+少突胶质细胞数量分别为(12.48±0.63)×10~6和(12.59±1.96)×10~6,而对照组28月龄雌、雄性大鼠大脑白质内CNPase~+少突胶质细胞数量分别为(5.05±1.31)×10~6和(6.48±0.84)×10~6,跑步训练组白质内CNPase~+少突胶质细胞数量显著较对照组28月龄多(p<0.01)。跑步训练组白质内大鼠CNPase+少突胶质细胞数量能够保持在对照组18月龄大鼠的水平(11.08±1.35)×10~6(p>0.05)和(11.65±1.32×10~6)(p>0.05)。 结论1.长期跑步训练对老年大鼠空间学习记忆能力具有积极的保护作用。2.长期跑步训练能够有效地减少髓鞘的退行性改变,并能显著降低雌、雄性大鼠大脑白质体积、白质内有髓神经纤维总长度和总体积以及少突胶质细胞数量的老年改变。长期跑步训练能够使28月龄雄性大鼠白质保持在对照组18月龄大鼠大脑白质各相关参数的水平,甚至使28月龄雌性大鼠白质各相关参数高于对照组18月龄大鼠白质的相关参数。 目的探讨长期跑步训练对大鼠大脑白质毛细血管老年改变的作用,以期进一步认识长期跑步训练对老年大脑白质毛细血管保护程度和机制。 方法跑步训练组、对照组28月龄和对照组18月龄每组随机挑选雄雌SD大鼠各5只,每只大鼠随机抽取一侧大脑半球,取出抽取的每侧大脑半球的白质,经Isector法冰冻包埋,获取5μm各向切片,通过免疫组织化学标记血管胶原Ⅳ。在放大1000倍下获取视野进行体视学分析,得出毛细血管的总长度、总表面积和总体积。 结果1.跑步训练组,雌、雄大鼠白质内毛细血管的总长度分别为36.92±4.65m和33.68±3.91m。对照组28月龄雌、雄大鼠白质内毛细血管的总长度分别为22.55±5.39m和18.42±2.55m,对照组18月龄雌、雄大鼠白质内毛细血管的总长度分别为29.31±2.97m和30.76±5.46m。跑步组雌性大鼠大脑白质内毛细血管总长度分别较对照组28月龄和对照组18月龄雌性大鼠白质内毛细血管总长度长63.73%(p0.01)和25.96%(p0.01);跑步组雄性大鼠大脑白质内毛细血管总长度分别较对照组28月龄和对照组18月龄雄性大鼠白质内毛细血管总长度长82.84%(p0.01)和9.49%(p0.05)。2.跑步训练组雌、雄大鼠白质内毛细血管的总表面积分别为6.35±0.99cm~2和8.79±1.22cm~2。对照组28月龄雌、雄大鼠白质内毛细血管总表面积分别为4.29±0.59cm~2和3.34±0.51cm~2,对照组18月龄时雌、雄大鼠白质内毛细血管的总表面积分别为8.08±1.96cm~2和6.79±1.53cm~2。在雌性大鼠,跑步训练组大脑白质内毛细血管总表面积分别较对照组28月龄和对照组18月龄白质内毛细血管总表面积大48.02%(p0.01)和21.41%(p0.05);在雄性大鼠,跑步训练组大脑白质内毛细血管总表面积分别较对照组28月龄和对照组18月龄白质内毛细血管总表面积大163.17%(p0.01) and29.46%(p0.05)。3.跑步训练组雌、雄大鼠白质内毛细血管的总体积分别为0.66±0.14mm~3和0.83±0.20mm~3。对照组28月龄雌、雄大鼠白质内毛细血管的总体积分别为0.69±0.24mm~3和1.06±0.29mm~3,对照组18月龄、雌雄大鼠白质内毛细血管的总体积分别为0.51±0.23mm~3和1.03±0.37mm~3。在雌性大鼠,跑步组大脑白质内毛细血管总体积分别较对照组28月龄和对照组18月龄白质内毛细血管总体积大37.74%(p0.05)和4.35%(p0.05);在雄性大鼠,,跑步组大脑白质内毛细血管总体积分别较对照组28月龄和对照组18月龄白质内毛细血管总体积大62.75%(p0.05)和19.42%(p0.05)。 结论长期跑步训练对老年大脑白质内毛细血管总长度、总体积和总表面积具有积极的保护作用。28月龄跑步训练组雄性大鼠大脑白质内毛细血管的各项参数显著性高于对照组28月龄雄大鼠大脑白质内毛细血管的各项参数,与18月龄对照组大鼠大鼠大脑白质内毛细血管的各项参数之间无显著性差异。28月龄跑步训练组雌性大鼠大脑白质内毛细血管的各项参数显著性高于对照组28月龄和对照组18月龄雌性大鼠大脑白质内毛细血管的各项参数。 目的探讨长期跑步锻炼对老年大鼠外周血和白质内神经、血管生长相关因子的作用,以期进一步揭示长期跑步训练对白质保护作用的可能机制。 方法从跑步训练组、对照组28月龄中每组随机挑选雄性SD大鼠各5只。在跑步完成后半小时内取外周血和大脑。白质在冰上分离后匀浆。血液和白质匀浆液离心后取上清液,用BCA法和Elisa试剂盒检测外周血和白质中神经血管相关因子含量。 结果1.跑步组外周血中的BDNF的含量是154.0±8.7ng/L,较对照组的124.9±10.3ng/L有显著性升高(p<0.01)。外周血中与神经、血管生长相关其余因子(FGF、IGF-1和NGF)跑步组和对照组之间没有显著差异(p>0.05)。2.对照组大鼠大脑白质中神经生长相关的因子BDNF、NGF、GDNF和NOGO-A的含量分别为675.2±93.9pg/mg、371.6±48.0pg/mg、543.8±67.1pg/mg和925.4±131.2pg/mg;跑步组大鼠大脑白质内神经生长的因子BDNF、NGF、GDNF和NOGO-A的含量分别为466.3±59.2pg/mg、295.7±79.4pg/mg、171.6±44.2pg/mg和334.9±61.3pg/mg,跑步训练组分别较对照组低30.9%(p㩳0.01)、50.8%(p㩳0.01)、45.6%(p㩳0.01)和63.8%(p㩳0.01)。跑步组大脑白质中血管形成相关的因子VEGF和Ang-1分别比对照组低52.1%(p㩳0.01)和51.3%(p㩳0.01)。 结论1.长期跑步训练促进了外周血中BDNF的显著性升高,使大脑白质中神经、血管生长相关因子的蛋白含量显著性降低。2.长期跑步训练对于对外周血和大脑白质中相关神经生长因子和相关血管形成因子的作用与长期跑步训练对老年大脑白质及老年大脑白质内毛细血管保护作用的关系尚需进一步的深入研究。
[Abstract]:Stereological study of the changes of the first part of long - term running on the aged changes in the brain white matter of rats with myelinated nerve fibers and less glial cells
Objective To explore the role of medium - intensity running training for middle - intensity running training for the aged 14 months to change the aged changes of cerebral white matter and myelinated nerve fibers and microglial cells , with a view to understanding the morphological basis and mechanism of long - term running exercise to protect the white matter of the brain .
Methods : Female and male SD rats were randomly selected for 4 months ( 14 months ) , running for 4 months ( 18 months ) and end of running exercise ( 28 months ) .
Results 1 . After 4 months of running training , the latency of male rats was significantly shorter than that before running ( p & lt ; 0.05 ) , and the latency of female and male rats after running for 14 months showed no significant difference ( p > 0.05 ) .
There was no significant difference ( p > 0.05 ) between the incubation period after long - term exercise and the incubation period of 28 - month - old in the control group ( p > 0.05 ) . The proportion of nerve fibers with degeneration of brain white matter in the control group was 21.2 卤 11.04 % , 35.8 卤 9.78 % and 49.2 卤 2.56 % , respectively .
Compared with control group , the total length of myelinated nerve fibers was 45.48 % ( p & lt ; 0.01 ) and 34.83 % ( p & lt ; 0.01 ) .
Compared with the control group , the total length of myelinated nerve fibers was 23.52 % ( p < 0.05 ) . The total volume of myelinated nerve fibers was 55.98 % and 46.84 % ( p < 0.01 ) .
Compared with the control group , the number of CNPase + Oligodendrocytes in running group was ( 12.48 卤 0.63 ) 脳 10 ~ 6 and ( 12.59 卤 0.96 ) 脳 10 ~ 6 , while the number of CNPase ~ + Oligodendrocytes in the white matter of running group was significantly higher than that in control group ( 11.08 卤 1.35 ) 脳 10 ~ 6 ( p > 0.05 ) and ( 11.65 卤 1.32 脳 10 ~ 6 ) ( p > 0.05 ) .
Conclusion 1 . Long - term running training has a positive protective effect on the spatial learning and memory ability of old rats . Long - term running training can effectively reduce the degeneration of the myelin sheath and decrease the total length and total volume of myelinated nerve fibers in the white matter .
Objective To explore the effect of long - term running training on the changes of cerebral white capillary vessel in rats , with a view to further understanding the degree and mechanism of long - term running training for the aged cerebral white capillary vessel protection .
Methods All 5 male and female SD rats were randomly selected from each group at 28 months of age and 18 months of age in the control group . Each rat was randomly selected to extract the white matter from one side of the cerebral hemisphere , then the white matter of each cerebral hemisphere was taken out , then the slices were frozen and embedded by the Isector method , and the collagen 鈪
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