HIF-1α与TfR mRNA在脑白质软化动物模型中的表达及相关性研究
发布时间：2019-05-27 01:18
【摘要】：【目的】 建立新生4天SD大鼠脑室周围白质软化模型（periventricular lerkomalacia,PVL）。在构建PVL模型大鼠的条件下，研究HIF-1α对TfR mRNA表达的调节，为缺氧缺血脑病发生机制与脑铁代谢之间关系的研究提供实验基础依据。 【方法】 1．4日龄新生SD大鼠通过随机数字表随机分为两组：PVL组和对照组；PVL组采用分离结扎双侧颈总动脉并予缺氧的方法建立脑室周围白质软化新生大鼠动物模型；对照组仅分离双侧颈总动脉，不予结扎及缺氧。采用HE染色法观察脑组织病理学改变，Western blot法检测MBP蛋白表达，实验动物生长发育及神经行为学等方法对动物模型进行评价。 2．设立（1）对照＋HIF-1α组（2）对照＋空白组（3）PVL＋抗HIF-1α组（4）PVL＋PBS组；及（1）对照＋PBS组（2）对照＋抗HIF-1α组（3）PVL＋PBS组（4）PVL＋抗HIF-1α组；以Western blot技术测HIF-1α表达，以Real-time PCR技术测TfR mRNA的表达。 【结果】 1．对4日龄新生SD大鼠进行双侧颈总动脉结扎并予以缺氧后，模型组与对照组大鼠相比，模型组大鼠生长缓慢（P0.01），睁眼时间延迟（21.67±1.19dVS15.54±1.60d），悬崖逃避能力及双臂悬吊能力均降低，差异有统计学意义(P＜0.001)。在水迷宫实验中，模型组大鼠在水中寻找站台的游泳速度减慢、寻找站台的所用的时间延长、撤去平台后在平台所在象限停留时间缩短(P＜0.05)。 2．脑组织HE染色，模型组术后24小时、48小时可见弥散性皮质下和脑室周白质疏松水肿。 3．Western blot法显示，HI1周后模型组MBP表达量较对照组降低，差异具有统计学意义。 4．对照＋PBS组与PVL＋PBS组对比，Western blot法检测后者HIF-1α蛋白表达增加，Real-time PCR检测TfR mRNA的表达也增加，差异有统计学意义。 5．对照＋HIF-1α组与对照＋空白组对比，Western blot法检测前者HIF-1α表达增多，Real-time PCR检测TfR mRNA的表达也增多，差异有统计学意义。 6．PVL＋抗HIF-1α组与PVL＋PBS组对比，Western blot法检测后者HIF-1α表达增多，Real-time PCR检测TfR mRNA的表达也增多，差异有统计学意义。 【结论】 1．通过对4日龄新生SD大鼠进行双侧颈总动脉结扎联合缺氧的方法，可建立与人类早产儿脑室周围白质软化病理相似的PVL动物模型。 2．缺氧缺血PVL可见HIF-1α表达增多，TfR mRNA的表达也增多； 3．PVL模型中，抑制脑组织中HIF-1α的活性，，TfR mRNA表达量随之减少，增加HIF-1α的表达，TfR mRNA表达量随之增加。即可推断HIF-1α可诱导TfR mRNA表达增加。 4．TfR mRNA表达的增加，可能是脑缺氧缺血后铁代谢失衡的一个机制。由于脑铁代谢的复杂性，HIF-1α与TfR的作用机制需进一步研究。
[Abstract]:[objective] to establish a model of periventricular leukomalacia (periventricular lerkomalacia,PVL) in neonatal SD rats. Under the condition of constructing PVL model rats, the regulation of TfR mRNA expression by HIF-1 伪 was studied, which provided an experimental basis for the study of the relationship between the pathogenesis of hypoxia-ischemic encephalopathy and brain iron metabolism. [methods] 1.4-day-old neonatal SD rats were randomly divided into two groups: PVL group and control group, and the neonatal rat model of periventricular leukomalacia was established by separating and ligating bilateral common carotid arteries and hypoxia in PVL group. In the control group, only bilateral common carotid arteries were separated without ligation and hypoxia. The pathological changes of brain tissue were observed by HE staining. The expression of MBP protein was detected by, Western blot method, and the animal model was evaluated by growth and development of experimental animals and neurobehavior. 2. Set up (1) control + HIF- 1 伪 group (2) control + blank group (3) PVL+ anti HIF- 1 伪 group (4) PVL+PBS group; And (1) control + PBS group (2) control + anti-HIF- 1 伪 group (3) PVL+PBS group (4) PVL+ anti-HIF-1 伪 group, Western blot technique was used to detect the expression of HIF-1 伪, Real-time PCR technique was used to detect the expression of TfR mRNA. [results] 1. After bilateral common carotid artery occlusion and hypoxia in 4-day-old neonatal SD rats, the growth of the model group was slower than that of the control group (P 0.01). The time of eye opening was delayed (21.67 卤1.19dVS15.54 卤1.60d), and the cliff escape ability and double arm suspension ability were decreased (P < 0.001). In the water maze experiment, the swimming speed of the model group rats looking for the platform in the water slowed down, the time used to find the platform was prolonged, and the residence time in the quadrant of the platform was shortened after the platform was removed (P < 0.05). 2. HE staining of brain tissue showed disseminated subcortical and periventricular white matter loose edema 24 hours and 48 hours after operation in the model group. 3.Western blot method showed that the expression of MBP in the model group was lower than that in the control group after HI1 week, and the difference was statistically significant. 4. Compared with PVL+PBS group, the expression of HIF-1 伪 protein in control + PBS group and PVL+PBS group was increased by, Western blot method, and the expression of TfR mRNA detected by Real-time PCR was also increased, the difference was statistically significant. 5. the expression of HIF-1 伪 and TfR mRNA detected by, Western blot in control + HIF-1 伪 group and control + blank group were significantly higher than those in control + HIF-1 伪 group and control + blank group, respectively. 6. Compared with PVL+PBS group, the expression of HIF-1 伪 in PVL + anti-HIF-1 伪 group was increased by, Western blot method, and the expression of TfR mRNA in PVL + anti-PVL 伪 group was also higher than that in PVL + anti-PVL 伪 group, the difference was statistically significant. [conclusion] 1. PVL animal model can be established by bilateral common carotid artery occlusion combined with hypoxia in 4-day-old neonatal SD rats, which is similar to the pathology of periventricular leukomalacia in human premature infants. 2. The expression of HIF-1 伪 in PVL increased and the expression of, TfR mRNA increased in 3.PVL model. In 3.PVL model, the expression of HIF-1 伪 decreased and the expression of, TfR mRNA increased with the increase of HIF-1 伪 expression in brain tissue. It can be inferred that HIF-1 伪 can induce the increase of TfR mRNA expression. The increase of 4.TfR mRNA expression may be a mechanism of iron metabolism imbalance after cerebral hypoxia and ischemia. Due to the complexity of brain iron metabolism, the mechanism of interaction between HIF-1 伪 and TfR needs to be further studied.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332
本文编号：2485785
[Abstract]:[objective] to establish a model of periventricular leukomalacia (periventricular lerkomalacia,PVL) in neonatal SD rats. Under the condition of constructing PVL model rats, the regulation of TfR mRNA expression by HIF-1 伪 was studied, which provided an experimental basis for the study of the relationship between the pathogenesis of hypoxia-ischemic encephalopathy and brain iron metabolism. [methods] 1.4-day-old neonatal SD rats were randomly divided into two groups: PVL group and control group, and the neonatal rat model of periventricular leukomalacia was established by separating and ligating bilateral common carotid arteries and hypoxia in PVL group. In the control group, only bilateral common carotid arteries were separated without ligation and hypoxia. The pathological changes of brain tissue were observed by HE staining. The expression of MBP protein was detected by, Western blot method, and the animal model was evaluated by growth and development of experimental animals and neurobehavior. 2. Set up (1) control + HIF- 1 伪 group (2) control + blank group (3) PVL+ anti HIF- 1 伪 group (4) PVL+PBS group; And (1) control + PBS group (2) control + anti-HIF- 1 伪 group (3) PVL+PBS group (4) PVL+ anti-HIF-1 伪 group, Western blot technique was used to detect the expression of HIF-1 伪, Real-time PCR technique was used to detect the expression of TfR mRNA. [results] 1. After bilateral common carotid artery occlusion and hypoxia in 4-day-old neonatal SD rats, the growth of the model group was slower than that of the control group (P 0.01). The time of eye opening was delayed (21.67 卤1.19dVS15.54 卤1.60d), and the cliff escape ability and double arm suspension ability were decreased (P < 0.001). In the water maze experiment, the swimming speed of the model group rats looking for the platform in the water slowed down, the time used to find the platform was prolonged, and the residence time in the quadrant of the platform was shortened after the platform was removed (P < 0.05). 2. HE staining of brain tissue showed disseminated subcortical and periventricular white matter loose edema 24 hours and 48 hours after operation in the model group. 3.Western blot method showed that the expression of MBP in the model group was lower than that in the control group after HI1 week, and the difference was statistically significant. 4. Compared with PVL+PBS group, the expression of HIF-1 伪 protein in control + PBS group and PVL+PBS group was increased by, Western blot method, and the expression of TfR mRNA detected by Real-time PCR was also increased, the difference was statistically significant. 5. the expression of HIF-1 伪 and TfR mRNA detected by, Western blot in control + HIF-1 伪 group and control + blank group were significantly higher than those in control + HIF-1 伪 group and control + blank group, respectively. 6. Compared with PVL+PBS group, the expression of HIF-1 伪 in PVL + anti-HIF-1 伪 group was increased by, Western blot method, and the expression of TfR mRNA in PVL + anti-PVL 伪 group was also higher than that in PVL + anti-PVL 伪 group, the difference was statistically significant. [conclusion] 1. PVL animal model can be established by bilateral common carotid artery occlusion combined with hypoxia in 4-day-old neonatal SD rats, which is similar to the pathology of periventricular leukomalacia in human premature infants. 2. The expression of HIF-1 伪 in PVL increased and the expression of, TfR mRNA increased in 3.PVL model. In 3.PVL model, the expression of HIF-1 伪 decreased and the expression of, TfR mRNA increased with the increase of HIF-1 伪 expression in brain tissue. It can be inferred that HIF-1 伪 can induce the increase of TfR mRNA expression. The increase of 4.TfR mRNA expression may be a mechanism of iron metabolism imbalance after cerebral hypoxia and ischemia. Due to the complexity of brain iron metabolism, the mechanism of interaction between HIF-1 伪 and TfR needs to be further studied.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332
【参考文献】
相关期刊论文 前1条
1 李艳伟;赵晋英;黄俊;李琳;;脑缺血诱导大鼠皮层和海马二价金属离子转运体1表达增加的研究[J];神经解剖学杂志;2011年03期
本文编号：2485785
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