抑制p38MAPK信号通路调控癫痫大鼠脑组织中腺苷A1受体及ENT1表达的研究

发布时间：2018-07-05 04:39

本文选题：癫痫 + 腺苷A1受体　；参考：《遵义医学院》2017年硕士论文

【摘要】：目的:探索抑制p38MAPK信号通路后,癫痫大鼠的行为学和脑组织内腺苷A1受体及ENT1的表达变化。方法:健康成年雄性SD大鼠随机分为:正常对照组(n=5)、癫痫组(0h、24h、72h、1W、2W五个时间点,n=5×5)、SB203580组(n=5)及DMSO组(n=5)。建立氯化锂-匹鲁卡品癫痫大鼠模型,应用Racine评分标准观察各组大鼠首次癫痫发作潜伏期、1小时内癫痫发作次数;HE染色观察各组大鼠脑组织病理形态学改变;免疫组化、免疫荧光及Western blot检测腺苷A1受体在各组大鼠海马及颞叶新皮质中的表达情况;Western blot检测ENT1在各组大鼠海马中的表达情况。结果:1.行为学结果示:癫痫组的大鼠首次癫痫发作潜伏期为25.89±1.27 min,DMSO组为25.65±1.56 min,SB203580组为37.53±1.73 min;癫痫组的1小时内大鼠癫痫发作次数为7.4±1.07,DMSO组为7.0±0.83,SB203580组为3.0±0.70。大鼠首次癫痫发作潜伏期和1小时内癫痫发作次数在癫痫组及DMSO组之间比较,差异无统计学意义(P0.05);与上述两组比较,SB203580组大鼠首次癫痫发作潜伏期明显延长,1小时内癫痫发作次数减少,且差异具有统计学意义(P0.05)。2.HE染色结果示:正常对照组海马组织结构清晰,未见病理性改变;癫痫组海马组织可见大量神经细胞凋亡、坏死;SB203580组海马组织亦有部分神经细胞变性坏死,但程度较癫痫组轻。3.Western blot检测腺苷A1受体表达的结果示:正常对照组大鼠的海马及颞叶新皮质中腺苷A1受体有基础水平的表达;氯化锂-匹鲁卡品癫痫模型大鼠中,腺苷A1受体在癫痫造模成功后24h表达较正常对照组上调,72h表达达到高峰,与正常对照组相比,差异具有统计学意义(P0.05);在应用p38MAPK抑制剂SB203580干预后,癫痫造模后第72h,SB203580组大鼠腺苷A1受体的表达较癫痫组(72h)显著下调,差异具有统计学意义(P0.05);免疫组化检测腺苷A1受体表达的结果示:正常对照组大鼠海马组织腺苷A1受体阳性细胞数为5.33±1.45,癫痫组为25.33±3.18,SB203580组为12.33±1.45。与正常对照组相比,癫痫组腺苷A1受体的阳性细胞数明显增多,且着色增强,差异具有统计学意义(P0.05);与癫痫组相比,SB203580组的腺苷A1受体阳性细胞数减少,且着色较浅,差异具有统计学意义(P0.05)。免疫荧光检测腺苷A1受体表达的结果示:腺苷A1受体主要表达于神经元的胞膜及胞质中。4.Western blot检测ENT1表达的结果示:SB203580组(24h)大鼠海马组织中ENT1的表达较癫痫组(24h)显著下调,差异具有统计学意义(P0.05)。结论:p38MAPK抑制剂SB203580可减轻大鼠海马神经元病理损害,延长大鼠首次癫痫发作潜伏期和降低大鼠癫痫发作程度,并同时降低腺苷A1受体和ENT1的表达。
[Abstract]:Aim: to explore the changes of behavior and expression of adenosine A1 receptor and ENT1 in brain tissue of epileptic rats after inhibition of p38 MAPK signaling pathway. Methods: healthy adult male Sprague-Dawley rats were randomly divided into normal control group (n = 5), epilepsy group (n = 5) and DMSO group (n = 5). To establish the rat model of lithium-pilocarpine epilepsy, observe the number of epileptic seizures within 1 hour of the incubation period of the first seizure and observe the pathological and morphological changes of brain tissue of rats in each group by HE staining, and observe the pathological changes of brain tissue of rats in each group by using Racine scoring standard. The expression of adenosine A1 receptor in hippocampus and temporal lobe neocortex of rats was detected by immunofluorescence and Western blot. Western blot was used to detect the expression of ENT1 in hippocampus of rats. The result is 1: 1. The behavioral results showed that the latency of the first seizure was 25.89 卤1.27 min in DMSO group, 37.53 卤1.73 min in DMSO group (25.65 卤1.56 min SB203580), and 7.0 卤0.83SB203580 (3.0 卤0.70) in epileptic group within 1 hour. The latency of the first seizure and the number of seizures within 1 hour were compared between the epileptic group and the DMSO group. There was no significant difference between the two groups (P0.05); compared with the above two groups, the number of epileptic seizures in SB203580 group was significantly prolonged within 1 hour, and the difference was statistically significant (P0.05) .2.HE staining results showed that the hippocampal tissue structure of normal control group was clear. No pathological changes were observed in the hippocampus of epilepsy group, and a large number of neuronal apoptosis was observed in hippocampal tissue of epileptic group, and some nerve cells were denatured and necrotized in hippocampus tissue of group SB203580. The expression of adenosine A1 receptor in hippocampal and temporal lobe neocortex of normal control rats was detected by Western blot, and the expression of adenosine A1 receptor in hippocampal and temporal lobe neocortex of normal control group was detected by Western blot, and the expression of adenosine A1 receptor in rat model of epilepsy induced by lithium-pilocarpine was observed. The expression of adenosine A1 receptor reached its peak at 24 hours after epileptic modeling compared with the normal control group at 72 hours, and the difference was statistically significant (P0.05), after the treatment of p38 MAPK inhibitor SB203580, the expression of adenosine A1 receptor reached its peak at 72 h after treatment with p38 MAPK inhibitor SB203580. The expression of adenosine A1 receptor in SB203580 group was significantly lower than that in epileptic group (72 h). The expression of adenosine A1 receptor in hippocampus of normal control group was 5.33 卤1.45, and that of epileptic group was 25.33 卤3.18 SB203580 (12.33 卤1.45). Compared with the normal control group, the number of adenosine A1 receptor positive cells in epileptic group was significantly increased, the difference was statistically significant (P0.05), and the number of adenosine A1 receptor positive cells in SB203580 group was lower than that in epileptic group. The difference was statistically significant (P0.05). The expression of adenosine A1 receptor was mainly detected in the membrane and cytoplasm of neurons. 4. The expression of ENT1 was detected by Western blot. The results showed that the expression of ENT1 in hippocampus of rats in the group of W SB 203580 (24 h) was significantly lower than that in the group of epilepsy (24 h). The difference was statistically significant (P0.05). Conclusion SB203580, a inhibitor of p38 MAPK, can attenuate the pathological damage of hippocampal neurons, prolong the latency of first seizure and decrease the degree of epileptic seizure in rats, and decrease the expression of adenosine A1 receptor and ENT1 at the same time.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742.1

【相似文献】