Effects Of Jingqianping Granule On The Expression Of GABA_BR2 Of Anger-out Model Rats And Construction Of GABA_BR2 Eukaryotic Expression Vector

Objective: Our aim is to investigate the expression of gamma-aminobutyric acid (GABA) B receptor 2 in different brain regions of anger-out model rats, as well as to research the effects of Jingqianshu granule on the expression, and construction of GABABR2 eukaryotic expression vector.We can study the mechanisms of action of occurrence and pathogenesis of anger with the effect of Jingqianshu granule. At the same time, the construction of GABABR2 eukaryotic expression vector can provide material foundation for neuron in signal transduction.Methods: 1 We made the anger-out model rats by the method of social isolation plus resident-intruder. The model rat were treated with Jingqianshu granule, and evaluated by behavioral test. We analyzed the expression of GABABR2 in four different brain regions by the method of reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. 2 GABABR2 cDNA was amplified with mRNA from rat's brain as template by RT-PCR. The cDNA was cloned into plasmid PMD18-T, then digesting PMD18-T by restriction enzyme, and the products were cloned into eukaryotic expression plasmid pcDNA3.1-His-lacZ for constructing recombinant plasmid pcDNA3.1-His-lacZ- GABABR2. The accuracy of pcDNA3.1-His-lacZ-GABABR2 was confirmed by restrietion enzyme digestion and DNA sequencing.Results: 1 The behavioral test results: Compared with the normal group, the score of the model group rats display significant difference (P<0.05). Meanwhile, when treating with Jingqianshu graulne, the significant difference disappear .RT-PCR results: The mRNA expression of GABABR2 decreased obviously(P<0.01)in the four different brain regions comparing with the normal group. Compared with the model group, the Jingqianshu graulne group had significant improvement (P<0.01); All of them had no significant difference compared with normal group except in frontal region. Western blot findings: The expression of GABABR2 decreased obviously(P<0.01)in the four different brain regions comparing with the normal control group. Compared with the model group, the Jingqianshu graulne group had significant improvement(P<0.05). All of them had no significant difference compared with normal group. 2 By DNA sequeneing, the sequence of GABABR2 cDNA was crrect. By restriction enzymes digesting and DNA sequencing, recombinant plasmid pcDNA3.1-His-lacZ-GABABR2 was constructed correctly. Conclusion: The anger-out model rats can be prepared successfully by using the methods of resident intruder paradigm. The expressions changes of the GABABR2 in the four different brain regions are in close relation to occurrence and pathogenesis of anger. GABABR2 gene was cloned sueeessuflly, and eukaryotic expression vector pcDNA3.1-His-lacZ-GABABR2 was constructed successfully.