吡非尼酮对大鼠血液及脑脊液中神经元特异性烯醇化酶的影响
发布时间：2019-05-20 05:12
【摘要】：目的：吡非尼酮是近年来发现的抗纤维化新药，其抗纤维化作用效果明确，因而其在纤维化机制导致的慢性脑积水的治疗上有很大的应用前景，其安全性尚无论证，本研究通过测定喂食吡非尼酮后大鼠血清及脑脊液中神经元特异性烯醇化酶的含量，以观察其对神经的损伤作用。 方法： 1大鼠分组及处理：挑选清洁级SD健康成年雄性大鼠50只，体重220±10g，随机分为10组，每组5只，分为空白对照组、50mg/kg组、100mg/kg组、300mg/kg组、500mg/kg组、700mg/kg组、900mg/kg组、1100mg/kg组、1300mg/kg组、1500mg/kg组，各组大鼠分别在灌胃前1天、灌胃后1小时、6小时、12小时、24小时经眼眶采血，在灌胃后24小时采集脑脊液标本，采集脑脊液标本后均灌注取脑。 2大鼠灌胃：各组大鼠灌胃前均禁食12小时，用大鼠灌胃器抽取配制好的PFD混悬液，给予大鼠灌胃处理。 3大鼠行为学检测：各组大鼠于灌胃前一天、当天、后一天，分别用平衡木行走试验、转W试验对大鼠行为能力进行评分。 4血清标本取材：各大鼠于预定时间经大鼠眼眶后静脉丛取血标本，用ELISA法检测血清中NSE的含量。 5脑脊液标本取材：各大鼠于预定时间用枕大池穿刺法取脑脊液标本，用ELISA法检测脑脊液中NSE的含量。 6脑标本取材：各大鼠于预定时间点灌注后断头取脑，脑标本石蜡包埋后留存待检。 7染色处理：用镀银染色法观察大鼠脑皮层组织病理学改变。 8数据分析：应用IBM SPSS Statistics version21统计软件进行统计学分析。 结果： 1一般状态变化：空白组大鼠实验期间，一般状态良好，精神好，进食好，被毛有光泽，活动灵活，未见明显异常状态，体重无明显变化。实验组大鼠中，低剂量组大鼠一般状态与空白组大鼠情况相近，随剂量越大，大鼠反应越重，一般状态越差，精神萎靡、饮食摄水减少，活动减少，随着时间延长，状态渐恢复，但仍不及实验前表现。各组实验大鼠均未出现死亡情况。 2各组大鼠体重比较：各组体重灌胃后较实验前均有所下降，但各组前后体重差异均无统计学意义（P0.05）。其体重变化考虑为实验刺激应激所致。 3各组大鼠平衡木试验比较：各组大鼠行非参数性检验，灌胃前一天各组大鼠间比较均无统计学差异(P0.05)。在每一组的大鼠灌胃均有统计学显著差异（P＜0.05）。灌胃后一天各组大鼠间比较均有统计学差异(P0.05)。各组大鼠组内不同时相间进行比较，空白组、50mg/kg组、100mg/kg组大鼠不同时间点之间平衡木试验记分差异有统计学意义（P0.05），300mg/kg组、500mg/kg组、700mg/kg组、1100mg/kg组、1300mg/kg组、1500mg/kg组各组大鼠在不同时间点之间其平衡木实验记分差别均有统计学意义（P0.05）。 4各组大鼠转圈试验比较：各组大鼠记录结果用非参数检验进行分析，灌胃前一天各组大鼠之间差异没有统计学意义（P0.05）。灌胃当天和灌胃后一天，各组大鼠之间的比较，差别均有统计学意义（P0.05）。空白组、50mg/kg组、100mg/kg组、300mg/kg组、500mg/kg组大鼠分别比较不同时相间差异，其差异均无统计学意义（P0.05），700mg/kg组、900mg/kg组、1100mg/kg组、1300mg/kg组、1500组大鼠分别比较不同时相间差异，其差异均有统计学意义（P0.05）。 5各组大鼠脑组织重量及密度比较：各组大鼠脑组织重量（g）组间两两进行比较，大剂量组1100mg/kg组、1300mg/kg组、1500mg/kg组比其小剂量组50mg/kg组、100mg/kg组、300mg/kg组及空白组脑重量均小，且差异有统计学意义（P0.05）。各组大鼠脑组织体积（ml）组间两两进行比较，大剂量组1100mg/kg组、1500mg/kg组比小剂量组300mg/kg组及空白组脑体积小，差异有统计学意义（P0.05），其它组间比较差异无统计学意义（P0.05）。各组大鼠脑组织密度（g/ml）组间两两进行比较，差异无统计学意义（P0.05）。 6各组大鼠血清NSE比较：各组大鼠灌胃前一天血清中NSE含量组间两两进行比较差异没有统计学意义（P0.05）。各组大鼠灌胃后1小时血清中NSE含量，空白组、50mg/kg组、100mg/kg组、300mg/kg组之间差异没有统计学意义（P0.05），500mg/kg组与700mg/kg组之间差异没有统计学意义（P0.05），900mg/kg组与1100mg/kg组之间差异没有统计学意义（P0.05），1500mg/kg组与其它组之间两两进行比较差异有统计学意义（P0.05）。各组大鼠灌胃后6小时血清中NSE含量（ng/ml），空白组、50mg/kg组、100mg/kg组、300mg/kg组之间差异没有统计学意义（P0.05），其余组两两组间进行对比差别均有统计学意义（P0.05）。各组大鼠灌胃后12小时血清中NSE含量，空白组、50mg/kg组、100mg/kg组、300mg/kg组之间差异没有统计学意义（P0.05），900mg/kg组与1100mg/kg组之间差异没有统计学意义（P0.05），其余各组与之两两进行比较差异有统计学意义（P0.05）。各组大鼠灌胃后24小时血清中NSE含量，各组间两两进行比较差异有统计学意义（P0.05）。空白组、50mg/kg组、100mg/kg组、300mg/kg组大鼠各时间点血清中NSE含量差别没有统计学差异（P0.05）。500mg/kg组、700mg/kg组、1100mg/kg组大鼠各时间点血清中NSE含量相比较，6小时、12小时最高，1小时、24小时次之，灌胃前最低，差异有统计学意义（P0.05）。900mg/kg组大鼠各时间点血清中NSE含量相比较，12小时最高，6小时次之，1小时、24小时再次之，灌胃前最低，差异有统计学意义（P0.05）。1300mg/kg组大鼠各时间点血清中NSE含量相比较，各时间点差异有统计学意义（P0.05）。1500mg/kg组大鼠各时间点血清中NSE含量相比较，各时间点两两不相同，，差异有统计学意义（P0.05），由小到大依次为灌胃前、1小时、24小时、6小时、12小时。 7各组大鼠脑脊液NSE比较：各组大鼠灌胃后24小时脑脊液中NSE含量，各组间两两进行比较差异有统计学意义（P0.05）。将其与24小时血清中NSE含量进行比较，其含量要大，且差异有统计学意义（P0.05）。 8各组大鼠大脑镀银染色比较：空白对照组大鼠脑组织神经细胞数量及形态结构分布正常。给予灌胃PFD的大鼠，小剂量组（50mg/kg、100mg/kg、300mg/kg）其脑组织神经细胞数量及形态结构分布与正常大鼠相近，大剂量组（500mg/kg）大鼠脑组织神经可见坏死，随剂量增大，坏死程度加重，正常神经元结构减少。 结论： 1NSE可作为大鼠脑损伤的重要检测指标，且其含量与损伤程度有相关性； 2NSE在大鼠脑损伤后有其分布规律，脑脊液中含量要高于血清中含量； 3PFD对大鼠脑神经有一定损伤作用； 4PFD对大鼠脑神经损伤有其剂量规律，50mg/kg、100mg/kg、300mg/kg对大鼠脑神经的作用不明显，500mg/kg以及更大剂量对大鼠脑神经的损伤作用呈正相关性；
[Abstract]:Objective: To study the effect of unifenone in the treatment of chronic hydrocephalus which has been found in recent years, and its anti-fibrosis effect is clear, so it has a great application prospect in the treatment of chronic hydrocephalus caused by fibrosis mechanism. In this study, the content of neuron-specific enolase in the serum and cerebrospinal fluid of rats fed with non-nitrone was determined to observe the damage to the nerve. square Method:1 rat group and treatment:50 healthy adult male rats with clean-grade SD were selected, weighing 220 to 10 g, and randomly divided into 10 groups, each group was divided into a blank control group, a 50 mg/ kg group, a 100 mg/ kg group, a 300 mg/ kg group, a 500 mg/ kg group, a 700 mg/ kg group, a 900 mg/ kg group, a 1100 mg/ kg group, a 1300 mg/ kg group, and a 1500 mg/ kg group. The rats were divided into four groups:1 day,1 hour,6 hours,12 hours and 24 hours after the intragastric administration respectively. The samples of the cerebrospinal fluid were collected 24 hours after the intragastric administration, and the cerebrospinal fluid samples were collected and the samples were collected. injection of the brain. Rats were given intragastric administration: the rats were fasted for 12 hours before the intragastric administration, and the prepared PFD suspension was extracted with the rats in the rat, and the rats were given a large amount Rats were treated by intragastric administration.3 The behavior of rats: the day before and after the administration of the rats, the day of the day, the day after the administration of the rats, the balance wood walking test was used for each group, and the rats were treated with the W test. The ability was scored.4 serum samples: the rats were collected from the rat's orbit after the scheduled time, and the blood samples were collected by ELISA. The content of NSE in serum was measured in 5 CSF specimens. The content of NSE in the cerebrospinal fluid (CSF) was measured. and the paraffin embedded in the brain specimen is stored for the to-be-tested.7 dyeing treatment: dyeing with silver-plated Methods To observe the pathological changes of the cerebral cortex of the rat.8 Data analysis: The application of the IBM SPSS Statistics ver Sio Statistical analysis of n21 statistical software. Results:1 General status change: during the experimental period of the blank group rats, the normal condition was good, the spirit was good, the food was good, and the hair was lustrous. In the experimental group, the normal state of the rats in the low-dose group was similar to that of the blank group. The larger the dosage, the more the rats reacted, the worse the general state, the listlessness, the decrease of the diet and the decrease of the activity. The time is extended, the state is gradually restored, but it still does not And before and after the experiment, there was no death in each group of experimental rats. The weight of each group was compared with that of each group. However, there was no significant difference in body weight before and after each group ( (P0.05). The weight change of each group was considered to be due to the experimental stimulation of stress. There was no statistical difference between the rats in the day before and after the stomach (P0.05). There was a statistically significant difference in each group of rats (P <0.05). There was no statistical difference between each group of rats in the day after the stomach (P0.05). There was no significant difference in the scores of the balance wood between the groups in the group, the blank group, the 50 mg/ kg group and the 100 mg/ kg group (P0.05), the 300 mg/ kg group, the 500 mg/ kg group and the 700 mg/ kg group. Rats in the kg group,1100 mg/ kg group,1300 mg/ kg group and 1500 mg/ kg group at different time points There was a significant difference in the experimental scores of the balance wood in each group (P0.05). There was no significant difference in the difference between the rats in the previous day (P0.05). The day of intragastric administration and the day after the gastric administration There was no significant difference between the rats in the blank group, the 50 mg/ kg group, the 100 mg/ kg group, the 300 mg/ kg group and the 500 mg/ kg group, respectively. The difference was not significant (P0.05), the 700 mg/ kg group,900 mg/ kg group,1100 mg/ kg group,1300 mg/ kg group and 1500 rats were respectively The difference of brain tissue weight and density of each group was statistically significant (P0.05). The weight and density of brain tissue in each group were compared with that of the group. The weight and density of brain tissue in each group were compared with that of the group of the rats in the group of 100 mg/ kg,1300 mg/ kg and 1500 mg/ kg in the group of 50mg/ kg, 100mg/ kg and 300mg/ kg. The weight of the brain in the kg group and the blank group was small, and the difference was significant (P0.05). (P0.05). There was no significant difference between the other groups (P0.05). The density of brain tissue in each group (P0.05). There was no significant difference between the two groups (P0.05). There was no significant difference between the two groups (P0.05). The levels of NSE in serum, blank group,50 mg/ kg group,100 mg/ kg group and 300 mg/ kg group were not statistically significant (P0.05). There was no statistical difference between the group of 500 mg/ kg and the group of 700 mg/ kg (P0.05), and there was no statistical difference between the group of 900 mg/ kg and the group of 1100 mg/ kg (P0.05), and 1500 mg. There was no significant difference (P> 0.05) between the/ kg group and the other groups (P <0.05). The serum NSE content (ng/ ml), the blank group, the 50 mg/ kg group, the 100 mg/ kg group and the 300 mg/ kg group were not statistically significant (P0. There was no significant difference between the two groups (P0.05). The levels of NSE, blank,50 mg/ kg,100 mg/ kg and 300 mg/ kg in the group were not significant (P0.05), and there was no statistical difference between the group of 900 mg/ kg and 1100 mg/ kg (P0.05). 05) There was a significant difference between the two groups (P0.05). The levels of NSE in the serum of the rats in the blank group, the 50 mg/ kg group, the 100 mg/ kg group and the 300 mg/ kg group were not statistically different (P0.05). The NSE content in the serum of the rats at the time points in the 500 mg/ kg group, the 700 mg/ kg group and the 1100 mg/ kg group was not statistically significant (P0.05). The levels of NSE in the serum of the rats in the 900 mg/ kg group were the highest and the difference was statistically significant (P0.05). The level of NSE in the serum of the rats at the 900 mg/ kg group was the highest, followed by 6 hours,1 hour and 24 hours, and the difference was statistically significant (P0. The levels of NSE in the serum of the rats at all time points in the 1300 mg/ kg group were statistically significant (P0.05). The levels of NSE in the serum of the 1500 mg/ kg group were not the same at each time point, and the difference was statistically significant (P0.05). 5) The levels of NSE in the cerebrospinal fluid of each group were as follows:1 hour,24 h,6 h and 12 h from small to large. The level of NSE in the spinal fluid and the difference between the two groups were statistically significant (P0.05). It was compared with the 24-hour serum. The content of NSE was higher and the difference was significant (P0.05). The results showed that the number of nerve cells and the morphological structure of the rats in the blank control group were normal, and the number of nerve cells and the morphological structure of the rats were similar to those of the normal rats and the large-dose group (500 mg/ kg). ) Rats The neurovisible necrosis of the brain tissue increases with the increase of the dose, the degree of necrosis is increased, and the normal neuronal structure is reduced. Conclusion: 1NSE can be used as an important test index for brain injury in rats, and its content and damage degree of correlation; NSE in rat brain The distribution of the brain in rats was higher than that in the serum, and the 3-D PFD had a certain damage to the rat's cranial nerve. The dose of the pfd to the rat's cranial nerve was 50 mg/ kg,100 mg/ kg and 300 mg/ kg.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
本文编号：2481380
[Abstract]:Objective: To study the effect of unifenone in the treatment of chronic hydrocephalus which has been found in recent years, and its anti-fibrosis effect is clear, so it has a great application prospect in the treatment of chronic hydrocephalus caused by fibrosis mechanism. In this study, the content of neuron-specific enolase in the serum and cerebrospinal fluid of rats fed with non-nitrone was determined to observe the damage to the nerve. square Method:1 rat group and treatment:50 healthy adult male rats with clean-grade SD were selected, weighing 220 to 10 g, and randomly divided into 10 groups, each group was divided into a blank control group, a 50 mg/ kg group, a 100 mg/ kg group, a 300 mg/ kg group, a 500 mg/ kg group, a 700 mg/ kg group, a 900 mg/ kg group, a 1100 mg/ kg group, a 1300 mg/ kg group, and a 1500 mg/ kg group. The rats were divided into four groups:1 day,1 hour,6 hours,12 hours and 24 hours after the intragastric administration respectively. The samples of the cerebrospinal fluid were collected 24 hours after the intragastric administration, and the cerebrospinal fluid samples were collected and the samples were collected. injection of the brain. Rats were given intragastric administration: the rats were fasted for 12 hours before the intragastric administration, and the prepared PFD suspension was extracted with the rats in the rat, and the rats were given a large amount Rats were treated by intragastric administration.3 The behavior of rats: the day before and after the administration of the rats, the day of the day, the day after the administration of the rats, the balance wood walking test was used for each group, and the rats were treated with the W test. The ability was scored.4 serum samples: the rats were collected from the rat's orbit after the scheduled time, and the blood samples were collected by ELISA. The content of NSE in serum was measured in 5 CSF specimens. The content of NSE in the cerebrospinal fluid (CSF) was measured. and the paraffin embedded in the brain specimen is stored for the to-be-tested.7 dyeing treatment: dyeing with silver-plated Methods To observe the pathological changes of the cerebral cortex of the rat.8 Data analysis: The application of the IBM SPSS Statistics ver Sio Statistical analysis of n21 statistical software. Results:1 General status change: during the experimental period of the blank group rats, the normal condition was good, the spirit was good, the food was good, and the hair was lustrous. In the experimental group, the normal state of the rats in the low-dose group was similar to that of the blank group. The larger the dosage, the more the rats reacted, the worse the general state, the listlessness, the decrease of the diet and the decrease of the activity. The time is extended, the state is gradually restored, but it still does not And before and after the experiment, there was no death in each group of experimental rats. The weight of each group was compared with that of each group. However, there was no significant difference in body weight before and after each group ( (P0.05). The weight change of each group was considered to be due to the experimental stimulation of stress. There was no statistical difference between the rats in the day before and after the stomach (P0.05). There was a statistically significant difference in each group of rats (P <0.05). There was no statistical difference between each group of rats in the day after the stomach (P0.05). There was no significant difference in the scores of the balance wood between the groups in the group, the blank group, the 50 mg/ kg group and the 100 mg/ kg group (P0.05), the 300 mg/ kg group, the 500 mg/ kg group and the 700 mg/ kg group. Rats in the kg group,1100 mg/ kg group,1300 mg/ kg group and 1500 mg/ kg group at different time points There was a significant difference in the experimental scores of the balance wood in each group (P0.05). There was no significant difference in the difference between the rats in the previous day (P0.05). The day of intragastric administration and the day after the gastric administration There was no significant difference between the rats in the blank group, the 50 mg/ kg group, the 100 mg/ kg group, the 300 mg/ kg group and the 500 mg/ kg group, respectively. The difference was not significant (P0.05), the 700 mg/ kg group,900 mg/ kg group,1100 mg/ kg group,1300 mg/ kg group and 1500 rats were respectively The difference of brain tissue weight and density of each group was statistically significant (P0.05). The weight and density of brain tissue in each group were compared with that of the group. The weight and density of brain tissue in each group were compared with that of the group of the rats in the group of 100 mg/ kg,1300 mg/ kg and 1500 mg/ kg in the group of 50mg/ kg, 100mg/ kg and 300mg/ kg. The weight of the brain in the kg group and the blank group was small, and the difference was significant (P0.05). (P0.05). There was no significant difference between the other groups (P0.05). The density of brain tissue in each group (P0.05). There was no significant difference between the two groups (P0.05). There was no significant difference between the two groups (P0.05). The levels of NSE in serum, blank group,50 mg/ kg group,100 mg/ kg group and 300 mg/ kg group were not statistically significant (P0.05). There was no statistical difference between the group of 500 mg/ kg and the group of 700 mg/ kg (P0.05), and there was no statistical difference between the group of 900 mg/ kg and the group of 1100 mg/ kg (P0.05), and 1500 mg. There was no significant difference (P> 0.05) between the/ kg group and the other groups (P <0.05). The serum NSE content (ng/ ml), the blank group, the 50 mg/ kg group, the 100 mg/ kg group and the 300 mg/ kg group were not statistically significant (P0. There was no significant difference between the two groups (P0.05). The levels of NSE, blank,50 mg/ kg,100 mg/ kg and 300 mg/ kg in the group were not significant (P0.05), and there was no statistical difference between the group of 900 mg/ kg and 1100 mg/ kg (P0.05). 05) There was a significant difference between the two groups (P0.05). The levels of NSE in the serum of the rats in the blank group, the 50 mg/ kg group, the 100 mg/ kg group and the 300 mg/ kg group were not statistically different (P0.05). The NSE content in the serum of the rats at the time points in the 500 mg/ kg group, the 700 mg/ kg group and the 1100 mg/ kg group was not statistically significant (P0.05). The levels of NSE in the serum of the rats in the 900 mg/ kg group were the highest and the difference was statistically significant (P0.05). The level of NSE in the serum of the rats at the 900 mg/ kg group was the highest, followed by 6 hours,1 hour and 24 hours, and the difference was statistically significant (P0. The levels of NSE in the serum of the rats at all time points in the 1300 mg/ kg group were statistically significant (P0.05). The levels of NSE in the serum of the 1500 mg/ kg group were not the same at each time point, and the difference was statistically significant (P0.05). 5) The levels of NSE in the cerebrospinal fluid of each group were as follows:1 hour,24 h,6 h and 12 h from small to large. The level of NSE in the spinal fluid and the difference between the two groups were statistically significant (P0.05). It was compared with the 24-hour serum. The content of NSE was higher and the difference was significant (P0.05). The results showed that the number of nerve cells and the morphological structure of the rats in the blank control group were normal, and the number of nerve cells and the morphological structure of the rats were similar to those of the normal rats and the large-dose group (500 mg/ kg). ) Rats The neurovisible necrosis of the brain tissue increases with the increase of the dose, the degree of necrosis is increased, and the normal neuronal structure is reduced. Conclusion: 1NSE can be used as an important test index for brain injury in rats, and its content and damage degree of correlation; NSE in rat brain The distribution of the brain in rats was higher than that in the serum, and the 3-D PFD had a certain damage to the rat's cranial nerve. The dose of the pfd to the rat's cranial nerve was 50 mg/ kg,100 mg/ kg and 300 mg/ kg.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965
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