银染mRNA差异显示法分离黄芪当归肾脏保护作用相关基因

摘要

Objective To identify genes associated with the chronic progression of renal disease and astragalus and angelica (A&A)'s renal protective effects.Methods The technique of silver staining mRNA differential display (DD) was used to investigate changes of gene expression in normal, sclerotic and A&A treated sclerotic kidneys. We isolated genes differentially expressed during the progression of renal disease which could be normalized by A&A.Results Several genes related to A&A's protective effects were isolated and one of them was confirmed by Northern blot.Conclusion Silver staining mRNA differential display is a simple and effective technique for isolating differentially expressed genes. The isolated new gene may be related to the progression of chronic renal disease and contribute to A&A's protective effects.%目的分离与肾脏疾病慢性进展以及黄芪当归防治肾硬化相关的基因.方法以银染mRNA差异显示法对比正常肾脏、肾病综合征后硬化肾脏和黄芪当归治疗组肾脏基因表达,分离疾病状态下表达发生改变且能被黄芪当归治疗纠正的基因.结果分离、克隆了26个黄芪当归保护作用相关基因片段,随机选取8个进行Northern杂交验证.其中一个基因片段被证实在肾硬化过程中表达下调,黄芪当归治疗组其表达有所恢复.测序后同源性比较显示此为一个新基因.结论银染mRNA差异显示法是简便快捷地分离差异基因的方法.本实验分离的新基因可能参与了肾脏疾病慢性进展并与黄芪当归治疗作用有关.