摘要
目的 分离与肾脏疾病慢性进展相关的基因 ,以及黄芪、当归药物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以及黄芪、当归治疗组的肾脏基因表达 ,并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 2 6个黄芪、当归对肾脏具有保护作用的相关基因片段 ,随机选取 8个进行Northern杂交验证 ,其一被证实在肾硬化过程中表达下调 ;黄芪、当归治疗组其表达恢复。测序后同源性比较显示为一新基因。结论 银染mRNA差异显示法是简便快捷的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的慢性进展 ,并与黄芪、当归的治疗作用有关。
Objective To isolate genes associated with chronic progression of renal diseases and renoprotective effects of Astragalus and Angelica(A & A). Methods Chronic puromycin nephropathy, which is characterized by massive proteinuria and progressive glomerular sclerosis and tubulointerstitial fibrosis, was induced in SD rats by repeated peritoneal injections of puromycin.Rats were randomly divided into three groups: (1) normal control (n=7), (2) puromycin nephropathy without A&A treatment (n=7), (3) puromycin nephropathy A & A treatment (3 ml/d, n=7). After 12 weeks, serum, urine and renal tissue were collected for study. The technique of silver staining mRNA differential display (DD) was used to investigate the changes of gene expression in kidneys of normal and nephropathy with and without A&A treatment rats. Genes expressing differently during the progression of renal diseases were isolated and the progression could be normalized by A&A. Results Twenty-six candidate cDNA fragments were isolated with DD. Northern blot analysis of eight randomly chosen candidate cDNA fragments demonstrated one exhibiting reduced mRNA expression in the fibrogenic process but upregulated by A&A treatment. Besides kidney, the gene was also expressed in lung, heart, liver and spleen. Whether it takes part in fibrogenic processes of these organs needs further study. Homology analysis through BLAST revealed that this gene was a novel one. Conclusion Silver staining mRNA differential display is a simple and quick technique for isolating differently expressed genes. The downregulated expression of the isolated new gene may contribute significantly to the fibrogenic process and be associated with A&A′s protective effects.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2000年第10期677-681,共5页
Chinese Journal of Internal Medicine
基金
国家自然科学基金!资助项目 (39970929)
