不同时间窗介入运动训练对实验性脑出血大鼠认知功能及海马可塑性的影响
发布时间：2018-08-07 21:20
【摘要】：目的:初步探讨不同时间窗介入运动训练对实验性脑出血大鼠认知功能以及海马可塑性的影响,为临床治疗提供依据和指导。方法:1、造模及干预方法:向大鼠右侧尾状核内注射Ⅰ型胶原酶及肝素钠建立实验性脑出血大鼠模型,对照组大鼠相同部位注入等体积生理盐水代替胶原酶及肝素钠,造模后进行改良神经功能缺损评分(Modified Neurological Severity Score,m NSS),并于术后12h进行正电子发射计算机断层显像/计算机体层扫描(Positron Emission Tomography/Computed Tomography PET/CT)检查,评分≥8.5分且PET/CT检查确有血肿存在即为造模成功,并根据开始介入运动训练的时间不同将运动训练组分别分为24小时(24h)组、3天(3d)组、7天(7d)组,分别于造模成功后24h、3d、7d开始运动训练,训练总时间为3周,对照组及模型组不予运动训练。2、指标观察:(1)训练结束后分别对各组大鼠进行Y-迷宫试验以评估大鼠分辨学习能力及一次性被动回避反应试验以评估记忆保持能力;(2)利用HE染色法观察脑出血模型大鼠脑出血区组织病理学改变;(3)用免疫组织化学法检测脑出血模型大鼠右侧大脑组织海马CA3区内源性神经营养因子(Brain derived neurotrophic factor,BDNF)和白细胞介素-10(Interleukin 10,IL-10)的蛋白表达量,以平均光敏度值(Optical density,OD值)表示;(4)采用RT-PCR法检测脑出血模型大鼠右侧大脑组织海马CA3区bax和bcl-2的基因表达,以灰度值表示。3、统计学分析:本实验的数据处理使用的是IBM SPSS Statitistics 17.0软件进行分析,实验数据结果用的均数±标准差(?x±s)表示,各组大鼠组间检测指标IL-10、BDNF平均光密度值的比较以及bax m RNA、bcl-2 m RNA、bcl-2 m RNA/bax m RNA灰度比值的比较均采用单因素方差分析(one-way ANOVA)进行两两比较,以p0.05表示差异具有统计学意义。结果:1、学习记忆能力检测:运动训练组大鼠的分辨学习能力评分均明显低于模型组,24h组、3d组、7d组达到学会标准所需次数分别为74.25±10.10,66.25±9.67,77.50±10.88,模型组达到学会标准所需次数为104.92±12.25(p0.05);运动训练组中24h组和7d组的分辨学习能力评分无明显差异(p0.05),但均显著高于3d组的分辨学习能力评分(p0.05);运动训练组大鼠记忆保持能力评分均明显优于模型组,24h组、3d组、7d组的电击后步入潜伏期(step-through latency,STL)分别为272.7s,283.1s,242.3s,模型组大鼠的电击后STL为132.8s(p0.05);运动训练组中24h组和3d组的记忆保持能力评分无明显差异(p0.05),但均显著高于7d组的记忆保持能力评分(p0.05)。2、脑组织形态观察:对照组大鼠脑组织未见右侧尾状核有血肿形成;造模成功大鼠则可见右侧尾状核区有不规则出血区,呈弥漫性,且术后1天可见血肿周围脑组织肿胀明显,呈浅淡模糊区,而术后28天可见血肿基本吸收,留下一囊性腔隙,囊腔周围脑组织肿胀明显消退,运动训练组大鼠血肿吸收较模型组稍快。3、HE染色:模型组及运动训练组较对照组有典型的病理学改变。对照组大鼠右侧尾状核脑组织结构均匀,染色均一,细胞轮廓完整清晰,胞浆丰富,胞核居中,未见明显神经胶质细胞增生及新生血管生成。模型组大鼠病灶区血肿吸收不完全,周围组织疏松,可见不均匀分布、排列不齐且大小不均匀的空泡,伴有少量神经胶质细胞增生及新生血管生成。与模型组比较,24h运动训练组血肿吸收较完全,周围组织疏松程度稍改善,仍可见大小不等的空泡,并伴有明显神经胶质细胞增生及新生血管生成。与24h运动训练组相比,3d组及7d组血肿吸收完全,周围组织疏松程度明显改善,空泡明显减少,新生血管生成及胶质细胞增生程度基本一致。4、BDNF、IL-10蛋白的表达:(1)对照组大鼠右侧大脑组织海马CA3区BDNF蛋白表达量极少,模型组和各运动训练组BDNF蛋白表达量均较对照组显著增高(p0.05);3个运动训练组BDNF蛋白表达量均较模型组明显增高(p0.05);3个运动训练组中,3d组和24h组的BDNF蛋白表达量无明显差异(p0.05),但均明显高于7d组(p0.05);(2)对照组大鼠右侧大脑组织海马CA3区IL-10蛋白表达量极少,模型组和各运动训练组IL-10蛋白表达量均较对照组显著增高(p0.05);3个运动训练组IL-10蛋白表达量均较模型组明显增高(p0.05);3个运动训练组中以3d组的IL-10蛋白表达量最高,而24h组和7d组两组之间IL-10蛋白表达量无明显差异(p0.05);5、bax m RNA、bcl-2 m RNA的表达:(1)bax m RNA的表达:对照组大鼠右侧大脑组织海马CA3区bax基因表达量极少,模型组与各运动训练组bax基因表达量均较对照组明显增加(p0.05);模型组bax基因表达量较3个运动训练组增加更为明显(p0.05);3个运动训练组中,以7d组bax基因表达量增高最为明显(p0.05),24h组和3d组的bax基因表达量无明显差异(p0.05)。(2)bcl-2 m RNA的表达:对照组大鼠右侧大脑组织海马CA3区bcl-2基因表达量较低,模型组与各运动训练组bcl-2基因表达量均较对照组增加(p0.05);3个运动训练组bcl-2基因表达量较模型组增加更为明显(p0.05);3个运动训练组中以3d组的bcl-2基因表达量最高,其次为24h组,最后为7d组(p0.05)。(3)bcl-2 m RNA/bax m RNA:运动训练组bcl-2 m RNA/bax m RNA比值均较对照组与模型组明显增加(p0.05);模型组与对照组的bcl-2 m RNA/bax m RNA比值无明显差异(p0.05);3个运动训练组中以3d组bcl-2 m RNA/bax m RNA比值最高,其次为24h组,再次为7d组(p0.05)。结论:1、运动训练能显著改善脑出血模型大鼠的认知功能,且在生命体征平稳,无严重并发症,神经功能障碍症状没有继续进展的情况下,脑出血后3天开始介入运动训练对大鼠认知功能的改善最有益;2、运动训练改善脑出血模型大鼠认知功能的机制可能与促进脑出血模型大鼠海马CA3区BDNF蛋白、IL-10蛋白及Bcl-2 m RNA的表达,抑制Bax m RNA表达,抑制海马CA3区神经元凋亡有关。
[Abstract]:Objective: To explore the effects of different time window intervention training on cognitive function and hippocampal plasticity in experimental cerebral hemorrhage rats, and to provide basis and guidance for clinical treatment. Methods: 1, modeling and intervention methods: injection of type I collagenase and hepatin sodium into the right caudate nucleus of rats to establish experimental cerebral hemorrhage model in the control group. The same part of the rat was injected into the same volume of physiological saline instead of collagenase and heparin sodium. The improved neural function defect score (Modified Neurological Severity Score, m NSS) was made after the model, and the positron emission computed tomography / computer tomography (Positron Emission Tomography/Computed Tomography PET/CT) was performed in 12h after the operation. Examination, score more than 8.5 points and PET/CT examination confirmed the existence of hematoma was a successful model, and the exercise training group was divided into 24 hours (24h) group, 3 days (3D) group, 7 days (7D) group according to the time of beginning intervention exercise training. After the model success, 24h, 3D, 7d began to exercise training, the total training time was 3 weeks, the control group and the model group did not exercise. Training.2, index observation: (1) after the training, Y- labyrinth test was carried out to rats in each group to evaluate the learning ability of rats and one time passive avoidance response test in order to evaluate the retention ability of the memory. (2) the pathological changes in the hemorrhage area of cerebral hemorrhage model rats were observed by HE staining method; (3) the immunohistochemical method was used to detect the brain out of the rats. The protein expression of endogenous neurotrophic factor (Brain derived neurotrophic factor, BDNF) and interleukin -10 (Interleukin 10, IL-10) in the hippocampal CA3 region of the right brain tissue of the rat's right blood model was expressed by the mean photosensitivity (Optical density, OD), and (4) the hippocampus of the right brain tissue of the rat model of cerebral hemorrhage was detected by the RT-PCR method. The gene expression of region Bax and Bcl-2 expressed.3 with gray value. Statistical analysis: the data processing of this experiment was used to analyze the IBM SPSS Statitistics 17 software, and the average number + standard deviation (? X + s) used in the experimental data was expressed, the comparison of the detection indexes between the rats in each group, the comparison of the average optical density value of the BDNF, and the Bax M The comparison of Cl-2 m RNA/bax m RNA grayscale ratio was compared with single factor analysis of variance (one-way ANOVA). The difference was statistically significant with P0.05. Results: 1, learning and memory ability test: the score of discrimination learning ability of the exercise training group was lower than that of the model group, 24h group, 3D group, 7d group met the standard of learning. The number of 74.25 + 10.10,66.25 + 9.67,77.50 + 10.88 in the model group was 104.92 + 12.25 (P0.05). There was no significant difference between the group 24h and the 7d group in the exercise training group (P0.05), but all were significantly higher than that of the 3D group (P0.05), and the memory retention of the exercise training group. All the 24h, 3D and 7d groups were significantly better than the model group. The latency period (step-through latency, STL) was 272.7s, 283.1s, 242.3s, and STL was 132.8s (P0.05) after the electric shock of the model group, and there was no significant difference in the memory retention ability of the exercise group and the exercise group, but all were significantly higher than the memory retention ability of the group. Score (P0.05).2, brain tissue morphology observation: in the control group, the brain tissue of the control group did not have hematoma in the right caudate nucleus, and the successful rat model showed that there was an irregular bleeding area in the right caudate nucleus, and the swelling of the brain tissue around the hematoma was obvious on the 1 day after the operation, and the hematoma was basically absorbed on the 28 day after the operation, leaving one of the basic absorption of the hematoma after the operation. The cystoid lacunae and the swelling of the brain tissue around the capsule obviously subsided. The hematoma absorption in the exercise training group was slightly faster than the model group.3, HE staining: the model group and the exercise training group had a typical pathological change compared with the control group. The brain tissue structure of the right caudate nucleus in the control group was uniform, the staining was all clear, the cytoplasm was rich, and the nucleus was in the middle. No obvious glial cell proliferation and neovascularization were found in the model group. The hematoma in the lesion area of the model group was incomplete, the surrounding tissue was loose, the nonhomogeneous distribution, the inhomogeneous and uneven vacuoles were arranged, with a small amount of glial cell proliferation and neovascularization. Compared with the model group, the hematoma absorption in the 24h exercise training group was over. There was a slight improvement in the degree of looseness in the surrounding tissue. The vacuoles of different sizes were still visible, accompanied by obvious glial cell proliferation and neovascularization. Compared with the 24h exercise training group, the hematoma in the 3D group and the 7d group was absorbed completely, the degree of porosity in the surrounding tissue was obviously improved, the vacuoles decreased obviously, the neovascularization and the degree of glial cell proliferation were basic. The expression of.4, BDNF and IL-10 protein: (1) the expression of BDNF protein in the hippocampus CA3 area of the right brain tissue of the control group was very little. The expression of BDNF protein in the model group and the exercise training group was significantly higher than that in the control group (P0.05), and the expression of BDNF protein in the 3 exercise groups was significantly higher than that in the model group (P0.05); the 3 exercise training groups, 3D group and 24 were in 3 exercise training groups. There was no significant difference in the expression of BDNF protein in group H (P0.05), but obviously higher than that in group 7d (P0.05). (2) the expression of IL-10 protein in the hippocampus CA3 region of the right brain tissue of the control group was very few. The expression of IL-10 protein in the model group and the exercise training group was significantly higher than that in the control group (P0.05), and the expression of IL-10 protein in the 3 exercise group was more than that of the model group. Xian Zenggao (P0.05); the expression of IL-10 protein in group 3D was the highest in 3 exercise groups, but there was no significant difference in the expression of IL-10 protein between group 24h and 7d group (P0.05); 5, Bax m RNA, bcl-2 m,: (1) the expression of gene expression in the hippocampus of the right large brain tissue of the control group was very few, model group and exercise training The expression of Bax gene in the practice group was significantly higher than that in the control group (P0.05); the expression of Bax gene in the model group was more obvious than that in the 3 exercise group (P0.05); the highest expression of Bax gene in the 7d group was the most obvious in the 3 exercise training groups (P0.05), and the Bax gene expression of 24h and 3D groups was not significantly different (P0.05). (2) the expression of Bcl-2 The expression of bcl-2 gene in the hippocampal CA3 region of the right brain tissue of the rats was lower, and the expression of bcl-2 gene in the model group and the exercise training group increased (P0.05), and the bcl-2 gene expression in the 3 exercise groups was more obvious than that in the model group (P0.05); the bcl-2 gene expression in the 3D group was the highest in the 3 exercise group, followed by the 24h group. At last, 7d group (P0.05). (3) Bcl-2 m RNA/bax m RNA ratio in Bcl-2 m RNA/bax m RNA: sports training group increased significantly than that of the control group and the model group, and the ratio of the model group and the control group was not significant. 7d group (P0.05). Conclusion: 1, exercise training can significantly improve the cognitive function of cerebral hemorrhage model rats, and in the condition of stable life signs, no serious complications, and the symptoms of nerve dysfunction do not continue to progress, 3 days after cerebral hemorrhage, intervention exercise training is the most beneficial to improve the recognition function of rats; 2, exercise training to improve cerebral hemorrhage model. The mechanism of cognitive function of type rats may be related to promoting the expression of BDNF protein, IL-10 protein and Bcl-2 m RNA, inhibiting the RNA expression of Bax m and inhibiting the apoptosis of hippocampal CA3 region neurons in the hippocampus CA3 area of cerebral hemorrhage model rats.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.34
本文编号：2171380
[Abstract]:Objective: To explore the effects of different time window intervention training on cognitive function and hippocampal plasticity in experimental cerebral hemorrhage rats, and to provide basis and guidance for clinical treatment. Methods: 1, modeling and intervention methods: injection of type I collagenase and hepatin sodium into the right caudate nucleus of rats to establish experimental cerebral hemorrhage model in the control group. The same part of the rat was injected into the same volume of physiological saline instead of collagenase and heparin sodium. The improved neural function defect score (Modified Neurological Severity Score, m NSS) was made after the model, and the positron emission computed tomography / computer tomography (Positron Emission Tomography/Computed Tomography PET/CT) was performed in 12h after the operation. Examination, score more than 8.5 points and PET/CT examination confirmed the existence of hematoma was a successful model, and the exercise training group was divided into 24 hours (24h) group, 3 days (3D) group, 7 days (7D) group according to the time of beginning intervention exercise training. After the model success, 24h, 3D, 7d began to exercise training, the total training time was 3 weeks, the control group and the model group did not exercise. Training.2, index observation: (1) after the training, Y- labyrinth test was carried out to rats in each group to evaluate the learning ability of rats and one time passive avoidance response test in order to evaluate the retention ability of the memory. (2) the pathological changes in the hemorrhage area of cerebral hemorrhage model rats were observed by HE staining method; (3) the immunohistochemical method was used to detect the brain out of the rats. The protein expression of endogenous neurotrophic factor (Brain derived neurotrophic factor, BDNF) and interleukin -10 (Interleukin 10, IL-10) in the hippocampal CA3 region of the right brain tissue of the rat's right blood model was expressed by the mean photosensitivity (Optical density, OD), and (4) the hippocampus of the right brain tissue of the rat model of cerebral hemorrhage was detected by the RT-PCR method. The gene expression of region Bax and Bcl-2 expressed.3 with gray value. Statistical analysis: the data processing of this experiment was used to analyze the IBM SPSS Statitistics 17 software, and the average number + standard deviation (? X + s) used in the experimental data was expressed, the comparison of the detection indexes between the rats in each group, the comparison of the average optical density value of the BDNF, and the Bax M The comparison of Cl-2 m RNA/bax m RNA grayscale ratio was compared with single factor analysis of variance (one-way ANOVA). The difference was statistically significant with P0.05. Results: 1, learning and memory ability test: the score of discrimination learning ability of the exercise training group was lower than that of the model group, 24h group, 3D group, 7d group met the standard of learning. The number of 74.25 + 10.10,66.25 + 9.67,77.50 + 10.88 in the model group was 104.92 + 12.25 (P0.05). There was no significant difference between the group 24h and the 7d group in the exercise training group (P0.05), but all were significantly higher than that of the 3D group (P0.05), and the memory retention of the exercise training group. All the 24h, 3D and 7d groups were significantly better than the model group. The latency period (step-through latency, STL) was 272.7s, 283.1s, 242.3s, and STL was 132.8s (P0.05) after the electric shock of the model group, and there was no significant difference in the memory retention ability of the exercise group and the exercise group, but all were significantly higher than the memory retention ability of the group. Score (P0.05).2, brain tissue morphology observation: in the control group, the brain tissue of the control group did not have hematoma in the right caudate nucleus, and the successful rat model showed that there was an irregular bleeding area in the right caudate nucleus, and the swelling of the brain tissue around the hematoma was obvious on the 1 day after the operation, and the hematoma was basically absorbed on the 28 day after the operation, leaving one of the basic absorption of the hematoma after the operation. The cystoid lacunae and the swelling of the brain tissue around the capsule obviously subsided. The hematoma absorption in the exercise training group was slightly faster than the model group.3, HE staining: the model group and the exercise training group had a typical pathological change compared with the control group. The brain tissue structure of the right caudate nucleus in the control group was uniform, the staining was all clear, the cytoplasm was rich, and the nucleus was in the middle. No obvious glial cell proliferation and neovascularization were found in the model group. The hematoma in the lesion area of the model group was incomplete, the surrounding tissue was loose, the nonhomogeneous distribution, the inhomogeneous and uneven vacuoles were arranged, with a small amount of glial cell proliferation and neovascularization. Compared with the model group, the hematoma absorption in the 24h exercise training group was over. There was a slight improvement in the degree of looseness in the surrounding tissue. The vacuoles of different sizes were still visible, accompanied by obvious glial cell proliferation and neovascularization. Compared with the 24h exercise training group, the hematoma in the 3D group and the 7d group was absorbed completely, the degree of porosity in the surrounding tissue was obviously improved, the vacuoles decreased obviously, the neovascularization and the degree of glial cell proliferation were basic. The expression of.4, BDNF and IL-10 protein: (1) the expression of BDNF protein in the hippocampus CA3 area of the right brain tissue of the control group was very little. The expression of BDNF protein in the model group and the exercise training group was significantly higher than that in the control group (P0.05), and the expression of BDNF protein in the 3 exercise groups was significantly higher than that in the model group (P0.05); the 3 exercise training groups, 3D group and 24 were in 3 exercise training groups. There was no significant difference in the expression of BDNF protein in group H (P0.05), but obviously higher than that in group 7d (P0.05). (2) the expression of IL-10 protein in the hippocampus CA3 region of the right brain tissue of the control group was very few. The expression of IL-10 protein in the model group and the exercise training group was significantly higher than that in the control group (P0.05), and the expression of IL-10 protein in the 3 exercise group was more than that of the model group. Xian Zenggao (P0.05); the expression of IL-10 protein in group 3D was the highest in 3 exercise groups, but there was no significant difference in the expression of IL-10 protein between group 24h and 7d group (P0.05); 5, Bax m RNA, bcl-2 m,: (1) the expression of gene expression in the hippocampus of the right large brain tissue of the control group was very few, model group and exercise training The expression of Bax gene in the practice group was significantly higher than that in the control group (P0.05); the expression of Bax gene in the model group was more obvious than that in the 3 exercise group (P0.05); the highest expression of Bax gene in the 7d group was the most obvious in the 3 exercise training groups (P0.05), and the Bax gene expression of 24h and 3D groups was not significantly different (P0.05). (2) the expression of Bcl-2 The expression of bcl-2 gene in the hippocampal CA3 region of the right brain tissue of the rats was lower, and the expression of bcl-2 gene in the model group and the exercise training group increased (P0.05), and the bcl-2 gene expression in the 3 exercise groups was more obvious than that in the model group (P0.05); the bcl-2 gene expression in the 3D group was the highest in the 3 exercise group, followed by the 24h group. At last, 7d group (P0.05). (3) Bcl-2 m RNA/bax m RNA ratio in Bcl-2 m RNA/bax m RNA: sports training group increased significantly than that of the control group and the model group, and the ratio of the model group and the control group was not significant. 7d group (P0.05). Conclusion: 1, exercise training can significantly improve the cognitive function of cerebral hemorrhage model rats, and in the condition of stable life signs, no serious complications, and the symptoms of nerve dysfunction do not continue to progress, 3 days after cerebral hemorrhage, intervention exercise training is the most beneficial to improve the recognition function of rats; 2, exercise training to improve cerebral hemorrhage model. The mechanism of cognitive function of type rats may be related to promoting the expression of BDNF protein, IL-10 protein and Bcl-2 m RNA, inhibiting the RNA expression of Bax m and inhibiting the apoptosis of hippocampal CA3 region neurons in the hippocampus CA3 area of cerebral hemorrhage model rats.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R743.34
【参考文献】
相关期刊论文 前10条
1 吕顺丽;杨敏;;脑出血后认知功能障碍研究新进展[J];西南军医;2017年02期
2 刘宇明;邓燕华;许治强;梁燕玲;胡佳佳;林永强;;脑出血患者血清NSE、GFAP、BDNF水平变化与认知障碍相关性研究[J];脑与神经疾病杂志;2016年10期
3 李红兵;谭子虎;;血管性痴呆的超微病理学研究进展[J];中西医结合心脑血管病杂志;2016年16期
4 杨淑梅;;奥拉西坦辅助治疗脑卒中认知功能障碍的临床疗效观察[J];世界最新医学信息文摘;2016年28期
5 章田野;;评价胞磷胆碱钠片治疗急性脑出血的效果及其安全性[J];中国医药指南;2016年08期
6 吴晓丹;;老年脑卒中患者记忆力障碍及其危险因素[J];中国老年学杂志;2016年05期
7 陈炜;黄树武;高玉广;何青;何乾超;黄德庆;张元侃;刘泰;;胶原酶与自体血建立脑出血模型的比较研究[J];药学研究;2015年12期
8 安宁;杨印东;解锦鼎;杨晓帆;;神经肌肉电刺激对脑卒中患者生活能力及抑郁状态的影响分析[J];中国药物依赖性杂志;2015年06期
9 王慧霞;;脑卒中后痴呆的预防和治疗[J];临床医药文献电子杂志;2015年35期
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