腺苷A1受体基因敲除小鼠戊四氮致痫后大脑皮层蛋白组学的研究

摘要

目的 观察腺苷A1受体基因敲除小鼠在戊四氮致痫后大脑皮层蛋白质的表达变化,探讨腺苷A1受体的抗癫痫和脑保护作用机制.方法 采用PCR技术鉴定腺苷A1受体基因敲除小鼠与野生型小鼠,应用戊四氮(30 mg/kg)腹腔注射制作癫痫模型,利用蛋白组学双向电泳检测癫痫发作后大脑皮层蛋白质表达变化,选取部分差异点进行质谱分析.结果 点燃组癫痫模型点燃成功后24 h、1个月,分别与对照组小鼠相比,大脑皮层多种蛋白质表达差异具有统计学意义.腺苷A1受体基因敲除小鼠与野生型小鼠相比较,对照组两者之间与点燃组两者之间多种蛋白质表达差异具有统计学意义.癫痫模型点燃成功后腺苷A1受体基因敲除小鼠与野生型小鼠相比,大脑皮层内表达下调蛋白为谷氨酰胺合成酶、醛缩酶A及甘氨酸受体;表达上调蛋白为蛋白激酶C及3-磷酸甘油醛脱氢酶.结论腺苷A1受体可能通过改变大脑皮层内多种蛋白质表达发挥其抗癫痫和脑保护作用.%Objective To investigate the anti-epileptic mechanism of A1 adenosine receptor by observing the variation of protein in cerebral cortex during pentylenetetrazol(PTZ)-induced kindling in A1 adenosine receptor knockout mice. Methods The A1 adenosine receptor knockout mice and the wild type mice were identified by PCR. The mice were given PTZ(30 mg/kg) by intraperitoneal injection. The variations of protein in cerebral cortex of mice were observed by two-dimensional electrophore-sis(2-DK) ; the part of these disparate points was taken for the mass chromatographic analysis. Results There was significant difference in the variation of protein in cerebral cortex at 24 h and 1 month after the mice were induced in the kindle groups as compared with the control groups. There was significant difference in the variation of protein in cerebral cortex between the A1 adenosine receptor knockout mice and the wild type mice in the control and kindle groups. In the kindle groups, glutaminesyn-thetase,aklolase A and glycine receptor were down-regulated,and protein kinase C and glyceraldehyde-3-phosphate dehydrogen-ase were up-regulated in the A1 adenosine receptor knockout mice as compared with the wild type mice. Conclusion The anti-epileptic mechanism of the A1 adenosine receptor may be contributed to the changes of protein in cerebral cortex regulated by the A1 adenosine receptor.