腦缺血大鼠血清生長(zhǎng)因子的動(dòng)態(tài)變化及重組人生長(zhǎng)激素對(duì)血管性癡呆模型大鼠作用的研究
發(fā)布時(shí)間：2019-04-03 11:27
【摘要】：目的:觀察慢性腦缺血大鼠血清及腦組織生長(zhǎng)激素及血清生長(zhǎng)因子的動(dòng)態(tài)變化;以及重組人生長(zhǎng)激素(recombinant human growth hormone,rhGH)對(duì)血管性癡呆(vascular dementia,VD)大鼠血清生長(zhǎng)因子及學(xué)習(xí)記憶的影響,探討重組人生長(zhǎng)激素對(duì)VD腦保護(hù)作用的機(jī)制。方法:1、采用Morris水迷宮篩選學(xué)習(xí)記憶能力正常的雄性SD大鼠143只,隨機(jī)分入各實(shí)驗(yàn)組,采用間斷性結(jié)扎大鼠雙側(cè)頸總動(dòng)脈法制備慢性腦缺血所致VD大鼠模型,術(shù)后,給藥組頸部皮下注射rhGH,給藥28d后,進(jìn)行Morris水迷宮實(shí)驗(yàn),觀察各組大鼠學(xué)習(xí)記憶能力情況;2、在腦缺血后各時(shí)間點(diǎn)分別從相應(yīng)實(shí)驗(yàn)組選取8只大鼠,采用酶聯(lián)免疫吸附法(ELISA)檢測(cè)血清、皮質(zhì)和海馬組織勻漿中的血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)、胰島素樣生長(zhǎng)因子-1(insulin-likegrowthfactor-1,IGF-1)和生長(zhǎng)激素(growth hormone,GH)的水平變化;另一部分大鼠斷頭取腦制作腦組織石蠟切片,進(jìn)行HE染色和TUNEL凋亡染色,觀察海馬CA1區(qū)病理學(xué)變化和神經(jīng)元的凋亡程度,采用Image Pro Plus圖像處理軟件處理TUNEL凋亡染色圖片,統(tǒng)計(jì)IOD值。結(jié)果:1、腦缺血大鼠VEGF、IGF-1和GH活性水平測(cè)定分析,與正常組相比,模型組血清、皮質(zhì)及海馬vegf、igf-1及gh活性水平隨腦缺血時(shí)間延長(zhǎng)均降低且有顯著差異性(p0.05);并且血清、皮質(zhì)和海馬vegf、igf水平和gh水平之間存在正相關(guān)性;在he染色中,模型組大鼠隨缺血時(shí)間增加海馬ca1區(qū)細(xì)胞排列紊亂,分布稀疏,細(xì)胞形狀不規(guī)則,細(xì)核變形,出現(xiàn)核固縮、溶解,而正常組無此改變;2、在水迷宮實(shí)驗(yàn)中,隨著訓(xùn)練天數(shù)的增加,各組逃避潛伏期均縮短。定位航行實(shí)驗(yàn)第六天,假手術(shù)組、模型組(28d)、重組人生長(zhǎng)激素組(28d)逃避潛伏期分別為5.29±3.41s、23.45±3.58s和7.30±5.39s,空間探索實(shí)驗(yàn)穿越平臺(tái)次數(shù)分別為5.87±3.27、1.32±1.19、3.14±1.50,可以發(fā)現(xiàn):與正常組比較,模型組(28d)大鼠學(xué)習(xí)記憶能力顯著下降,差異有統(tǒng)計(jì)學(xué)意義(p0.05),說明模型制備成功。與模型組(28d)比較,重組人生長(zhǎng)激素組(28d)學(xué)習(xí)記憶能力顯著增加,差異有統(tǒng)計(jì)學(xué)意義(p0.05),較正常組差異無統(tǒng)計(jì)學(xué)意義。重組人生長(zhǎng)激素組大鼠vegf、igf-1和gh水平測(cè)定分析,與模型組相比,重組人生長(zhǎng)激素組血清、皮質(zhì)及海馬vegf、igf-1及gh活性水平增高且有顯著差異性(p0.05);tunel凋亡染色觀察發(fā)現(xiàn),與模型組比較,重組人生長(zhǎng)激素組海馬ca1區(qū)神經(jīng)元陽(yáng)性染色減少且分布密度有所增加;與模型組比較,重組人生長(zhǎng)激素組iod值降低,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。結(jié)論:慢性腦缺血導(dǎo)致血清、皮質(zhì)和海馬igf-1及vegf均減少,并且與血清、皮質(zhì)和海馬gh水平具有直線相關(guān)性。rhgh可以增加vd大鼠gh、igf-1及vegf水平,改善其學(xué)習(xí)記憶及認(rèn)知功能障礙,減輕海馬神經(jīng)元凋亡,提示rhgh對(duì)vd大鼠慢性缺血性腦損傷具有保護(hù)作用。
[Abstract]:Objective: to observe the dynamic changes of serum and brain growth hormone and serum growth factor in rats with chronic cerebral ischemia. And the effect of recombinant human growth hormone (recombinant human growth hormone,rhGH on serum growth factor and learning and memory in rats with vascular dementia (vascular dementia,VD), and to explore the mechanism of the protective effect of recombinant human growth hormone on the brain of VD. Methods: 1143 male SD rats with normal learning and memory ability were screened by Morris water maze and randomly divided into each experimental group. Chronic cerebral ischemia-induced VD rats were established by intermittent ligation of bilateral common carotid artery. Twenty-eight days after subcutaneous injection of rhGH, into the neck of the treatment group, Morris water maze test was performed to observe the learning and memory ability of the rats in each group. 2. At each time point after cerebral ischemia, 8 rats were selected from the corresponding experimental group, and the levels of vascular endothelial growth factor (vascular endothelial growth factor,VEGF in serum, cortex and hippocampus homogenate were detected by enzyme-linked immunosorbent assay (ELISA). Changes of insulin-like growth factor-1 (insulin-likegrowthfactor-1,IGF-1) and growth hormone (growth hormone,GH) levels; The other part of rats were decapitated to make paraffin sections of brain tissue, HE staining and TUNEL apoptosis staining were used to observe the pathological changes of hippocampal CA1 region and the degree of neuron apoptosis, and Image Pro Plus image processing software was used to process the images of TUNEL apoptosis staining. Statistical IOD value. Results: 1. The activity levels of VEGF,IGF-1 and GH in cerebral ischemia rats were measured and analyzed. Compared with normal group, vegf, in serum, cortex and hippocampus of model group were measured and analyzed. The activity levels of igf-1 and gh decreased with the prolongation of cerebral ischemia time and showed significant difference (p0.05). There was a positive correlation between the levels of vegf,igf and gh in serum, cortex and hippocampus. In he staining, with the increase of ischemia time, the cells in the ca1 region of hippocampus in the model group were disarranged and distributed sparsely, the shape of the cells was irregular, the fine nucleus was deformed, the nucleus was condensed and dissolved, but there was no such change in the normal group. 2. In the water maze test, the escape latency was shortened with the increase of training days. The escape latencies of sham operation group, model group (28 days) and recombinant human growth hormone group (28 days) were 5.29 鹵3.41 s, 23.45 鹵3.58 s and 7.30 鹵5.39 s, respectively. The times of crossing the platform in spatial exploration experiment were 5.87 鹵3.27,1.32 鹵1.19,3.14 鹵1.50respectively. It was found that the learning and memory ability of the model group (28d) was significantly lower than that of the normal group (p0.05), and there was a significant difference between the two groups (p0.05). The results showed that the model was prepared successfully. Compared with the model group (28d), the learning and memory ability of the recombinant human growth hormone group (28d) was significantly increased (p0.05), but there was no significant difference compared with the normal group (p0.05). The levels of vegf,igf-1 and gh in the recombinant human growth hormone group were higher than those in the model group. Compared with the model group, the levels of vegf,igf-1 and gh in the serum, cortex and hippocampus in the recombinant human growth hormone group were significantly higher than those in the model group (p0.05). Tunel apoptosis staining showed that compared with the model group, the positive staining of hippocampal ca1 neurons in the recombinant human growth hormone group decreased and the distribution density increased. Compared with the model group, the OD value of the recombinant human growth hormone group was significantly lower (p 0.05). Conclusion: chronic cerebral ischemia can decrease the levels of igf-1 and vegf in serum, cortex and hippocampus, and have linear correlation with serum, cortex and hippocampus gh levels. Rhgh can increase gh,igf-1 and vegf levels in vd rats. It is suggested that rhgh has protective effect on chronic ischemic brain injury in vd rats by improving learning, memory and cognitive impairment and alleviating apoptosis of hippocampal neurons.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R965
[Abstract]:Objective: to observe the dynamic changes of serum and brain growth hormone and serum growth factor in rats with chronic cerebral ischemia. And the effect of recombinant human growth hormone (recombinant human growth hormone,rhGH on serum growth factor and learning and memory in rats with vascular dementia (vascular dementia,VD), and to explore the mechanism of the protective effect of recombinant human growth hormone on the brain of VD. Methods: 1143 male SD rats with normal learning and memory ability were screened by Morris water maze and randomly divided into each experimental group. Chronic cerebral ischemia-induced VD rats were established by intermittent ligation of bilateral common carotid artery. Twenty-eight days after subcutaneous injection of rhGH, into the neck of the treatment group, Morris water maze test was performed to observe the learning and memory ability of the rats in each group. 2. At each time point after cerebral ischemia, 8 rats were selected from the corresponding experimental group, and the levels of vascular endothelial growth factor (vascular endothelial growth factor,VEGF in serum, cortex and hippocampus homogenate were detected by enzyme-linked immunosorbent assay (ELISA). Changes of insulin-like growth factor-1 (insulin-likegrowthfactor-1,IGF-1) and growth hormone (growth hormone,GH) levels; The other part of rats were decapitated to make paraffin sections of brain tissue, HE staining and TUNEL apoptosis staining were used to observe the pathological changes of hippocampal CA1 region and the degree of neuron apoptosis, and Image Pro Plus image processing software was used to process the images of TUNEL apoptosis staining. Statistical IOD value. Results: 1. The activity levels of VEGF,IGF-1 and GH in cerebral ischemia rats were measured and analyzed. Compared with normal group, vegf, in serum, cortex and hippocampus of model group were measured and analyzed. The activity levels of igf-1 and gh decreased with the prolongation of cerebral ischemia time and showed significant difference (p0.05). There was a positive correlation between the levels of vegf,igf and gh in serum, cortex and hippocampus. In he staining, with the increase of ischemia time, the cells in the ca1 region of hippocampus in the model group were disarranged and distributed sparsely, the shape of the cells was irregular, the fine nucleus was deformed, the nucleus was condensed and dissolved, but there was no such change in the normal group. 2. In the water maze test, the escape latency was shortened with the increase of training days. The escape latencies of sham operation group, model group (28 days) and recombinant human growth hormone group (28 days) were 5.29 鹵3.41 s, 23.45 鹵3.58 s and 7.30 鹵5.39 s, respectively. The times of crossing the platform in spatial exploration experiment were 5.87 鹵3.27,1.32 鹵1.19,3.14 鹵1.50respectively. It was found that the learning and memory ability of the model group (28d) was significantly lower than that of the normal group (p0.05), and there was a significant difference between the two groups (p0.05). The results showed that the model was prepared successfully. Compared with the model group (28d), the learning and memory ability of the recombinant human growth hormone group (28d) was significantly increased (p0.05), but there was no significant difference compared with the normal group (p0.05). The levels of vegf,igf-1 and gh in the recombinant human growth hormone group were higher than those in the model group. Compared with the model group, the levels of vegf,igf-1 and gh in the serum, cortex and hippocampus in the recombinant human growth hormone group were significantly higher than those in the model group (p0.05). Tunel apoptosis staining showed that compared with the model group, the positive staining of hippocampal ca1 neurons in the recombinant human growth hormone group decreased and the distribution density increased. Compared with the model group, the OD value of the recombinant human growth hormone group was significantly lower (p 0.05). Conclusion: chronic cerebral ischemia can decrease the levels of igf-1 and vegf in serum, cortex and hippocampus, and have linear correlation with serum, cortex and hippocampus gh levels. Rhgh can increase gh,igf-1 and vegf levels in vd rats. It is suggested that rhgh has protective effect on chronic ischemic brain injury in vd rats by improving learning, memory and cognitive impairment and alleviating apoptosis of hippocampal neurons.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R965
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