康欣口服液对老年小鼠脑线粒体DNA缺失突变影响的实验研究

Experimental Study on the Effect of KangXin oral solution on brain mitochondrial DNA deletion mutation in aged Balb/c mice

目的:研究康欣口服液对Balb/c小鼠脑线粒体DNA(mtDNA)缺失突变的影响。方法:近交系Balb/c小鼠,分为青年组(6周),中年组(6个月),老年组(14个月)各10只,提取小鼠脑组织的mtDNA,采用聚合酶链反应(PCR)技术扩增缺失型的mtDNA片段,以证实老年组小鼠脑组织中存在mtDNA缺失。近交系Balb/c老年小鼠分别灌胃给予生理盐水(老年空白对照组),康欣口服液(老年康欣口服液组),连续四个月后处死,提取脑组织的mtDNA,PCR技术分别扩增野生型和缺失型的mtDNA片段,运用凝胶成像仪进行光密度扫描,比较两组缺失型mtDNA/野生型mtDNA的光密度比值。结果:近交系老年Balb/c小鼠脑mtDNA中存在明显的片段缺失,而在中年组与青年组中未检测到相同片段的缺失。与老年空白对照组相比,康欣口服液能显著降低老年Balb/c小鼠mtDNA的缺失率(P<0.001)。结论:mtDNA的缺失突变随年龄而积累,康欣口服液可以抑制老年小鼠脑mtDNA的缺失。

Objective： To explore the effect of KangXin oral solution on brain mitochondrial DNA （mtDNA） deletion mutation in aged Balb/c mice. Methods： The Balb/c mice were divided into the young group （6weeks）, middle-aged group （6months） and aged group （14 months）, each group has 10 mice. Brain mtDNA were obtained and polymerase chain reaction （PCR） technique was used to examine the fragment deletion of brain mtDNA, thus, to confirm there was deletion of mtDNA in aged mice, s brain. The aged Balb/c mice were divided into two groups： the aged blank control group being given 0.9% normal sodium, KangXin oral solution group. After being treated for four months, the brain mtDNA were obtained and polymerase chain reaction （PCR） technique was used to amplify wild-type and deletion from of mtDNA. Gel imaging meter was used to detect optical density, then, to compare the optical density ratios of deletion from mtDNA/wide type mtDNA in two groups. Results： There were 304bp mtDNA deletion in brain mitochondrial of aged Balb/c mice, but same mtDNA deletions were not detected in brain mitochondrial of young and middle-aged mice. Compared with aged blank control group, the mtDNA deletion of aged Balb/c mice in KangXin oral solution group decreased obviously （P〈0.001）. Conclusion： mtDNA deletion mutation accumulates with the increase of age. KangXin oral solution can inhibit mtDNA deletion of aged mice.