异亚丙基莽草酸对慢性低灌注大鼠海马CA1区胆碱乙酰转移酶表达水平的影响  

Influence of 3,4-oxo-isopropylidene-shikimic acid on expression of choline acetyltransferase in CA1 subfield of hippocampus in rats with chronic hypoperfusion

作　　者：王晶[1] 周宁家[1] 张硕峰[1] 方芳[1] 孙建宁[1] 

机构地区：[1]北京中医药大学中药学院中药药理系,北京100102

出　　处：《北京中医药大学学报》2008年第8期529-533,577,共6页Journal of Beijing University of Traditional Chinese Medicine

基　　金："十五"国家科技攻关项目(No.2001BA701A07-14);国家留学基金资助留学研究生项目(No.2006100796)

摘　　要：目的探讨慢性低灌注大鼠海马CA1区胆碱乙酰转移酶(ChAT)的蛋白表达以及基因转录水平的改变,并观察异亚丙基莽草酸(ISA)对其影响。方法实验动物分为假手术组、模型组、喜得镇组、ISA100 mg/kg组、ISA50 mg/kg组以及ISA25 mg/kg组。将大鼠双侧颈总动脉结扎加切断后8周,经心灌注固定,制成大脑石蜡切片后进行免疫组织化学染色,测定海马CA1区ChAT定位以及相对定量表达。抽提大鼠海马CA1区组织总RNA,通过荧光定量实时PCR技术,对ChATmRNA表达水平进行测定。结果慢性低灌注65 d后,大鼠海马CA1区ChAT免疫反应阳性产物主要分布于锥体细胞的胞浆及轴突。模型组大鼠的ChAT蛋白表达水平以及ChATmRNA表达水平非常显著低于假手术组(P<0.05)。与模型组相比,各给药组的ChAT蛋白表达水平不同程度升高(P<0.05);ISA100 mg/kg组的ChATmRNA表达水平增加非常显著(P<0.05),而ISA其余各剂量以及喜得镇对ChATmRNA表达水平无显著影响(P>0.05)。结论ISA可增加慢性低灌注大鼠海马CA1区ChAT的蛋白以及基因表达水平。Objective To investigate the changes of expression and genetic transcription of choline acetyhransferase （CHAT） in CA1 subfield of hippocampus in rats with chronic hypoperfusion, and observe the influence of 3, 4-oxo-isopropylidene-shikimic acid （ISA） on these changes. Methods The rats were divided into the sham-operation group, model group, Hydergine group, ISA group 1 （100 mg/ kg）, ISA group 2 （50 mg/kg） and ISA group 3 （25 mg/kg）. The brain paraffin sections were stained by immunohistochemistry method after bilateral carotid ligation and amputation for 8 weeks and heart perfusion and fixation in rats. The ChAT position and relevant quantitative expression in CA1 subfield of hippocampus were determined. The total mRNA of CA1 subfield of hippocampus in rats was extracted and the expression of ChAT mRNA was detected by polymerase chain reaction （PCR） of fluorescent quantitative real-time technique. Results After chronic hypoperfusion for 65 days the positive products of immuno-reaction in CA1 subfield of hippocampus mostly distributed in pyramidal neuron plasma and axons. In the model group the protein expressions of ChAT and ChAT mRNA were much lower than those in the sham-operation group （P 〈 0.05 ）, while those in different ISA groups increased in varying degrees （P 〈 0.05） compared with those in the model group. In ISA group 1 the expression of ChAT mRNA increased very significantly （P 〈 0.05 ）. In other two ISA groups and Hydergine group the expression of ChAT mRNA had no obvious changes （P 〉 0.05）. Conclusion ISA can increase the protein and gene expressions of ChAT in CA1 subfield of hippocampus in rats with chronic hypoperfusion.

关 键 词：异亚丙基莽草酸 慢性低灌注 海马 胆碱乙酰转移酶 大鼠 

分 类 号：R285.5[医药卫生—中药学]