Basic studies for an effective therapy for Alport syndrome

1.用Col 4a 4基因缺陷小鼠进行药物治疗Alport综合征的研究与未治疗组和咪唑立宾治疗组相比，环孢素A治疗组和依那普利治疗组的蛋白尿和新月体形成明显减少。依那普利能显著延长突变小鼠的生存期，而环孢素A和咪唑立宾则不能。总之，血管紧张素转换酶抑制剂被认为在Alport综合征小鼠模型中有效减缓向肾衰竭的进展。Col 4a 4、tenascin双缺陷小鼠的分析与Col 4a 4缺陷小鼠相比，Col 4a 4、tenascin双缺陷小鼠的尿蛋白和生存期明显改善。Col 4a 4基因缺陷小鼠肾小球基底膜tenascin基因表达缺失可能参与了Alport综合征的病理改变.利用Col 4a 4转基因小鼠对Col 4a 4基因缺陷小鼠表型的挽救研究将小鼠与Col 4a 4缺陷小鼠交配。我们现在正在研究Col 4a 4缺陷小鼠的表型是否被拯救。电穿孔法基因治疗Alport综合征的研究将质粒pCAG-Col 4a 4-LacZ经腹主动脉或肾包膜电穿孔至小鼠肾脏。虽然经腹主动脉导入DNA的效率很低，但相当数量的肾皮质细胞通过电穿孔从包膜下途径导入质粒。包膜下电穿孔法可用于Alport综合征等肾脏疾病的基因治疗。

1. Study for the efficiency of drug therapy for Alport syndrome using Col4a4 deficient miceCompared with untreated and mizoribine-treated groups, proteinuria and crescent formation were significantly reduced in cyclosporine A-treated and enalapril-treated groups. Furthermore, enalapril could significantly prolong the survival period of mutant mice, although cyclosporine A and mizoribine could not. Taken together, angiotensin converting enzyme inhibitor was thought to be effective in slowing down the progression towards renal failure in a mouse model of Alport syndrome.2. Analysis of Col4a4, tenascin double deficient miceCompared with Col4a4 deficient mice, proteinuria and survival period were significantly improved in Col4a4, tenascin double deficient mice. Elavation of tenascin gene expression observed at the glomerular basement membrane of Col4a4 deficient mice was thought to play a role for promote the pathological change of Alport syndrome.3. Rescue study of the phenotype of Col4a4 deficient mice by Col4a4 transgenic miceCol4a4 transgenic mice carrying CAG promoter and LacZ reporter gene were produced. The mice were mated with Col4a4 deficient mice. We are now examining whether the phenotype of Col4a4 deficient mice is rescued or not.4. Study of gene therapy for Alport syndrome using electroporationThe plasmid pCAG- Col4a4-LacZ was electroporated into mouse kidneys by either abdominal aorta route or renal capsule route. Although efficiency of DNA introduction via abdominal aorta was very low, considerable number of cells in renal cortex were introduced the plasmid by electroporation from subcapsular route. The electroporation from subcapsular route may be available for gene therapy for Alport syndrome and other renal diseases.