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目的探讨高热惊厥(febrile seizures,FS)对发育期大鼠脑内γ氨基丁酸B受体(γ- aminobutyric acid B receptor,GABABR)亚基GABABR1与GABABR2表达及结合改变的远期影响。方法采用热水浴诱导大鼠反复惊厥。隔日诱导惊厥1次,共诱导10次。发育期大鼠随机分为正常对照组(n=64)和高热处理组(n=268)。正常对照组大鼠置于37℃水中,高热处理组大鼠置于44．8℃热水中。高热处理组大鼠按其是否出现惊厥又分为高热对照组(H,n=64)和高热惊厥组。隔日诱导惊厥1次,共10次。惊厥组大鼠取惊厥1次(FS1,n=64)和10次(FS10,n=64)者作为实验对象。采用免疫双标记检测GABABR1与GABABR2共位点表达的改变;采用竞争RT—PCR检测GABABR1与GABABR2定量表达的改变;采用免疫共沉淀／Western Blot检测GABABR1与GABABR2结合的改变。结果(1)10次高热处理后,大鼠海马GABABR1和GABABR2的表达明显降低。(2)H组和FS1组大鼠GABABR1和GABABR2表达量及二者结合量在末次处理后7-15 d恢复正常,而FS10组未恢复正常。(3)FS10组在末次处理后30 d GABABR1表达恢复正常,而GABABR2及二者结合量未恢复正常。结论发育期大鼠反复高热惊厥可导致GABABR亚单位GABABR1与GABABR2表达及二者结合量的远期改变。
Objective To investigate the long-term effects of febrile seizures (FS) on the expression and binding changes of GABABR1 and GABABR2 in the brain of developing GABABR subunits in rats. Methods Hot water bath was used to induce repeated convulsions in rats. Convulsion induction every other day, a total of 10 induction. Developmental rats were randomly divided into normal control group (n = 64) and hyperthermia group (n = 268). Normal control rats were placed in water at 37 ℃, high heat treatment rats were placed in 44.8 ℃ hot water. High heat-treated rats were divided into high fever control group (H, n = 64) and febrile seizure group according to whether they had seizures or not. Convulsion induction every other day, a total of 10 times. The seizure group took convulsions once (FS1, n = 64) and 10 times (FS10, n = 64) as experimental subjects. Immunoblotting was used to detect the change of co-site expression of GABABR1 and GABABR2. The changes of quantitative expression of GABABR1 and GABABR2 were detected by competitive RT-PCR. The co-localization of GABABR1 and GABABR2 was detected by co-immunoprecipitation and Western Blot. Results (1) After high heat treatment for 10 times, the expression of GABABR1 and GABABR2 in hippocampus of rats decreased significantly. (2) The expression of GABABR1 and GABABR2 in group H and group FS1 and the combination between the two groups returned to normal at 7-15 d after the last treatment, while the group at FS10 did not return to normal. (3) The expression of GABABR1 in FS10 group returned to normal 30 days after the last treatment, but the combination of GABABR2 and the two did not return to normal. Conclusion The development of rats with recurrent febrile seizures can lead to the GABABR subunits GABABR1 and GABABR2 expression and long-term changes in the combination of the two.