少突胶质前体细胞在癫痫发生中的作用研究

发布时间：2018-06-05 08:58

本文选题：癫痫发生 + 脱髓鞘　；参考：《重庆医科大学》2014年博士论文

【摘要】：第一部分癫痫发生不同时期髓鞘及少突胶质细胞的动态变化目的：探索大鼠癫痫发生不同时期海马内髓鞘及少突胶质细胞的动态变化。方法：选用成年SD大鼠106只,分为正常对照组16只，实验组90只。实验组采用氯化锂-匹罗卡品腹腔注射诱导惊厥持续状态（StatusConvulsion,SC），利用视频联合无线遥测脑电监测SC后癫痫发生不同时期脑电图变化;采用免疫荧光染色检测SC后癫痫发生不同时期大鼠海马髓鞘蛋白MBP的形态变化；免疫荧光染色成熟少突胶质细胞标记物CC1,观察成熟少突胶质细胞数目变化；western blot测定SC后癫痫发生不同时期海马内髓鞘蛋白MBP及CNPase含量变化。结果：(1)大鼠SC后不同时期脑电图改变符合癫痫发生不同时期脑电图改变；(2)大鼠海马MBP、CNPase表达在SC后癫痫发生的过程中呈现动态变化，在癫痫形成的急性期MBP、CNPase表达即开始下降(P0.01)，并于SC后癫痫发生潜伏期、慢性期及慢性晚期均显著低于正常对照组，并呈进行性下降趋势（P0.01）；（3）大鼠SC后癫痫发生急性期CC1阳性细胞数目开始下降(P0.01)，SC后癫痫发生潜伏期、慢性期及慢性晚期均显著低于正常对照组（P0.01），与MBP、CNPase呈一致性变化。结论：SC后癫痫发生不同时期髓鞘及少突胶质细胞持续受损，该损伤贯彻于癫痫发生整个过程。第二部分少突胶质前体细胞在SC后癫痫发生中的作用目的：探索大鼠癫痫发生过程中少突胶质前体细胞及其调控因子的变化。方法：选用成年SD大鼠，实验组采用氯化锂-匹罗卡品腹腔注射诱导惊厥持续状态，应用视频联合无线遥测脑电监测大鼠SC后自发性发作情况。利用免疫荧光染色技术检测NG2蛋白表达情况，了解OPC在癫痫发生不同时期的动态变化；western blot测定PDGFR-α、olig1，olig2在癫痫发生不同时期的动态变化，了解OPC及调控OPC分化的关键转录因子在癫痫发生中的变化情况。结果：（1）OPC在SC后癫痫发生急性期开始活化增值，在癫痫发生潜伏期、癫痫发生慢性期显著高于正常对照组(P0.01)，在癫痫发生慢性晚期低于正常对照组(P0.01)。（2）转录因子olig1、olig2在SC后癫痫发生急性期开始升高(P0.01)，癫痫发生潜伏期达到峰值，随后在癫痫发生慢性期急剧下降，低于正常对照组(P0.01)，，持续到慢性晚期，维持在较低水平。结论：大鼠SC后癫痫发生早期出现OPC的活化，增值能力增强，可能具有修复髓鞘受损的作用；调控OPC增值分化的转录因子olig1、olig2在SC后癫痫发生早期上调可能促进OPC增值分化，参与髓鞘内源性修复，而在慢性期的急剧下降可能是导致髓鞘持续受损的原因之一。第三部分地塞米松在癫痫发生过程中的作用目的：探索地塞米松是否可以通过干预SC后癫痫发生过程中的髓鞘损伤，影响癫痫形成。方法：选用成年SD大鼠68只，随机分为正常对照组，DEX对照组，及实验组。实验组采用氯化锂-匹罗卡品腹腔注射诱导惊厥持续状态，SC90分钟后存活的大鼠再随机分为DEX干预组与模型组；DEX干预组于SC90分钟后腹腔给予地塞米松0.5mg/Kg.d,连用4天；DEX对照组采用等量的地塞米松腹腔注射。利用视频联合无线遥测脑电检测SC后各组大鼠癫痫自发性发作情况。在癫痫发生的慢性期采用免疫荧光染色检测各组海马区MBP表达情况；western blot测定MBP、CNPase、PDGFR-α蛋白含量。结果：地塞米松可以延长大鼠癫痫发生的静默期（P0.05），减少大鼠慢性期SRS次数（P0.05）；地塞米松对正常大鼠髓鞘、成熟少突胶质细胞及OPC无明显影响（P0.05）；地塞米松可以提高癫痫发生慢性期大鼠海马髓鞘蛋MBP含量（P0.05）；地塞米松可以降低慢性期大鼠海马PDGFR-α蛋白含量（P0.05）、提高CNPase蛋白含量（P0.05）。结论：地塞米松可能通过促进大鼠海马OPC分化为成熟少突胶质细胞，促进髓鞘修复,进而发挥其在氯化锂-匹罗卡品诱导的癫痫发生过程的抗痫作用。
[Abstract]:Dynamic Changes of myelin and Oligodendrocyte in the First Part of Epilepsy

Objective : To explore the dynamic changes of myelin sheath and microglial cells in hippocampus of rats during different periods of epilepsy .

Methods : 106 adult SD rats were divided into normal control group ( n = 16 ) and experimental group ( 90 rats ) .
The number of mature microglial cells was observed by immunofluorescence staining .
The content of MBP and CNPase in hippocampus of hippocampus after SC was determined by western blot .

Results : ( 1 ) EEG changes in different periods after SC were consistent with EEG changes in different periods of epilepsy .
( 2 ) The expression of MBP and CNPase in hippocampus of rats was dynamically changing after SC . In the acute stage of epilepsy , the expression of MBP and CNPase began to decrease ( P0.01 ) .
( 3 ) After SC , the number of positive cells of CC1 was decreased ( P0.01 ) , and the latency , chronic stage and late stage of epilepsy after SC were significantly lower than that of the normal control group ( P0.01 ) .

Conclusion : After SC seizures , the myelination and microglia cells continued to be damaged in different periods , and the damage was carried out in the whole process of epilepsy .

Role of Oligodendrocyte precursor cells in the pathogenesis of epilepsy after SC

Objective : To explore the changes of glial precursor cells and their regulatory factors in rats with epilepsy .

Methods : Adult SD rats were induced by intraperitoneal injection of lithium chloride - pilocarpine in the experimental group . The spontaneous seizures in rats were monitored with video combined with wireless telemetry . The expression of NG2 protein was detected by immunofluorescence staining technique , and the dynamic changes of OPC in different periods of epilepsy were investigated .
The dynamic changes of PDGFR - 伪 , bcl - 1 and bcl - 2 during different stages of epilepsy were measured by western blot . The changes of the key transcription factors in the development of epilepsy were investigated .

Results : ( 1 ) In the acute phase of epilepsy , the time of epilepsy onset was significantly higher than that of the normal control group ( P0.01 ) .

Conclusion : The activation of OPC and the enhancement of value - added ability of post - SC epilepsy may have the effect of repairing myelin damage .
The early - regulation of post - SC seizures may promote the differentiation of OPC value - added differentiation , participate in the endogenic repair of myelin sheath , and the acute decrease in chronic phase may be one of the causes of the continued damage of myelin sheath .

The Role of Dexamethasone in the Process of Epilepsy

Objective : To explore whether dexamethasone can affect the formation of epilepsy by intervening the myelin sheath injury in the process of post - SC seizure .

Methods : 68 adult SD rats were randomly divided into normal control group , DEX control group and experimental group .
After SC90 minutes , dexamethasone 0.5 mg / kg 路 d was intraperitoneally administered to DEX group .
DEX control group was injected intraperitoneally with an equal amount of dexamethasone . In the chronic phase of epilepsy , the expression of MBP was detected by immunofluorescence staining .
The contents of MBP , CNPase and PDGFR - 伪 were determined by western blot .

Results : Dexamethasone could prolong the silent period of epilepsy in rats ( P0.05 ) , and reduce the number of SRS ( P0.05 ) .
Dexamethasone has no significant effect on normal rat myelin sheaths , mature microglial cells and OPC ( P0.05 ) .
Dexamethasone could increase the content of MBP in hippocampus of rats with chronic epilepsy ( P0.05 ) .
Dexamethasone could reduce the content of PDGFR - 伪 protein in hippocampus of rats with chronic stage ( P0.05 ) , and increase the content of CNPase protein ( P0.05 ) .

Conclusion : Dexamethasone may play an important role in epilepsy induced by lithium chloride - pilocarpine by promoting the differentiation of OPC into mature microglial cells and promoting the repair of myelin sheath .
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R742.1

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