高浓度腺苷对脑缺血再灌注线粒体编码基因表达的影响

The effects of high concentration Adenosine on the expression of gene coded by mitchondia after reperfusion

目的针对线粒体的半自主特性,采用高浓度腺苷对脑缺血再灌注线粒体自体编码基因表达进行干预,探讨高浓度腺苷对缺血再灌注神经元能量代谢环节的影响。方法颈动脉分流法制备大鼠脑缺血再灌注模型,提取各实验组脑线粒体,采用Fernandez-Silva的细胞器体外3H-UTP掺入法建立脑线粒体体外RNA转录体系,根据最新线粒体基因序列进行引物设计,用PCR仪对RNA转录体系产物进行PCR扩增及定性、定量分析。结果在腺苷浓度2mmol/L的情况下线粒体编码基因-Cox Ⅰ mRNA、Cox Ⅱ mRNA、ATPase6 mRNA的相对表达量与缺血再灌注组的Cox Ⅰ、Cox Ⅱ、Cox Ⅲ、mRNA相对表达量相比均有明显降低(P<0.01,P<0.05,P<0.05)。在腺苷浓度2mmol/L的情况下线粒体编码基因-Cox ⅢmRNA、ATPase8 mRNA相对表达量与缺血再灌注后相对表达量差别无统计学意义(P>0.05)。结论脑缺血再灌注后,过高浓度的胞外腺苷会导致线粒体自身编码基因的不正常表达,可能会导致脑能量代谢障碍加重。

Objective Focus on the half-self character of mitchondia, we use high concentration adenosine interfering the expression of gene coded by mitchondia to study high concentration adenosine, s effects on brain cell energy metabolism. Methods Using the methods of crotid shunt make models of rat brain isehemia and reperfusion, using the methods of Fernandez-Silva build RNA synthesis system. According to Human mitochondrial gene serial design primer, using the methods of PCR, examine the expression of gene Cox Ⅰ , Ⅱ, Ⅲ and ATPase 6,ATPase9 coded by mitochondria. Results The expression of Cox Ⅰ ,Cox Ⅱ ,ATPase6 with condition adenosine 2 mmol/L descend comparing with reperfusion, s （ P 〈 0.01, P 〈 0.05, P 〈 0.05 ）. The expression of Cox m, ATPase8 with condition adenosine 2 mmoL/L has no change comparing with reperfusion, s （ P 〉 0.05）. Conclusion High concentration adenosine can results abnormal exression of gene coded by mitchondia, perhaps, big dose adenosine aggrevate brain injury.