免疫组化、Western blot蛋白印迹和免疫荧光染色观察磷酸鞘氨醇受体2大鼠海马中的表达变化  

Immunohistochemistry, Western blotting and immunofluorescence staining to observe the expression of sphingosine phosphate receptor 2 in rat hippocampus

作　　者：王柳毅[1] 王乐[1] 董治燕[1] 钟红平[2] Wang Liuyi;Wang Le;Dong Zhiyan;Zhong Hongping(Department of Neurology,the Second Affiliated Hospital of Yanan University,Shanxi 716000,China)

机构地区：[1]延安大学附属医院神经内科,陕西716000 [2]延安大学附属医院小儿科,陕西716000

出　　处：《脑与神经疾病杂志》2020年第2期101-104,共4页Journal of Brain and Nervous Diseases

摘　　要：目的观察磷酸鞘氨醇受体2(S1PR2)基因在颞叶癫痫大鼠海马中的表达变化。方法将72只健康雄性Wistar大鼠随机均分为实验组(36例)和对照组(36例)。实验组使用氯化锂—匹罗卡品进行颞叶癫痫诱导制模,成功后持续大发作1h终止待检,对照组以相同剂量生理盐水注射,然后以进行免疫组化、Western blot印迹和免疫荧光染色检测并进行统计学分析。结果免疫组化检测结果显示,两组1d、3d、7d、28d、56d海马S1PR2阳性细胞计数相比有统计学意义(P<0.05);Western blot检测结果显示,两组3d、7d、28d、56d海马S1PR2蛋白表达水平相比有统计学意义(P<0.05);免疫荧光染色检测显示,S1PR2蛋白表达阳性为红色,星形胶质细胞为绿色,细胞核为蓝色。结论颞叶癫痫大鼠海马中S1PR2表达水平显著下降,说明颞叶癫痫发病机制可能与神经元功能、星形胶质细胞增生受到S1PR2影响有关。Objective To observe the expression of sphingosine phosphate receptor 2(S1PR2)in hippocampus of temporal lobe epilepsy rats.Methods 72 healthy male Wistar rats were randomly divided into experimental group(36 cases)and control group(36 cases).In the experimental group,lithium chloride-pilocarpine was used to induce temporal lobe epilepsy and the model was terminated for 1 hour after the successful onset.The control group was injected with the same dose of saline,and then detected by immunohistochemistry,Western blotting and immunofluorescence staining,and statistical analysis was carried out.Results Immunohistochemical results showed that there was significant difference in the number of hippocampal S1PR2 positive cells between the two groups on the 1st,3rd,7th,28th and 56th day(P<0.05).Western blotting results showed that the expression of S1PR2 protein in hippocampus of the two groups on the 3rd,7th,28th and 56th days was significantly different(P<0.05).Immunofluorescence staining showed that the positive expression of S1P2 was red,astrocyte was green and nucleus was blue.Conclusions The expression of S1PR2 in hippocampus of rats with temporal lobe epilepsy decreased significantly,suggesting that the pathogenesis of temporal lobe epilepsy may be related to the influence of S1PR2 on neuron function and astrocyte proliferation.

关 键 词：磷酸鞘氨醇受体2 海马 免疫组化 Western blot 免疫荧光染色 

分 类 号：R742.1[医药卫生—神经病学与精神病学]