Basic Studies on Gene Therapy for Glomerulonephritis

1)遗传性肾脏疾病的遗传分析我们分析了Alport综合征患者的基因组DNA，其特征是肾小球基底膜的特殊形态变化，如篮状编织外观。我们已经从3例患者中鉴定出IV型胶原α5链缺失。良性家族性血尿又称薄基底膜病，以肾小球基底膜致密层弥漫性变薄为特征。我们已经进行了IV型胶原α3或α4链与…的遗传连锁分析，但没有连锁，提示与TBMD相关的基因不是这些基底膜胶原。2)大鼠实验性肾小球肾炎的转录因子。我们重点研究了原癌基因c-fos、c-jun和c-myc，以及作为TBMD的异源二聚体复合体AP-1在抗Thy 1，1肾炎和链脲佐菌素糖尿病大鼠中，c-Fos和c-jun的表达增加，c-myc的表达在抗Thy 1，1肾炎后4天或5天达到高峰，这与系膜细胞的增殖相一致。AP-1复合体在STZ糖尿病大鼠慢性肾小球分泌期有较强的表达，而不是在糖尿病诱导后表达。这些结果表明，这些转录因子在系膜增生性肾小球肾炎中起重要作用，特别是在疾病的进展期和慢性期。3)全反式维甲酸(ATRA)对系膜细胞的影响ATRA被认为是细胞分化的诱导剂。这可能表明全反式维甲酸通过诱导系膜细胞表型改变而抑制系膜细胞的生长。全反式维甲酸抑制肾小球系膜细胞生长及c-fos、c-jun和c-myc基因表达。然后用全反式维甲酸治疗抗TH1.1肾炎。ATRA处理的大鼠肾小球系膜细胞数量和Matix评分明显受到抑制。这些结果有力地表明，这些转录因子的下调可能是治疗系膜增生性肾小球肾炎的候选药物。较少

1) Genetic analysis of hereditary renal diseasesWe have analyzed the genomic DNA from Alport syndrome patients, which is characterized by specific morphological changes of glomerular basement membrane such as basket weave appearance. We have identified the deletion of type IV collagen alpha 5 chain from 3 patients. Two cases had partial deletion and another was supposed to have complete deletion of the alpha 5 and alpha 6 chains.Benign familial hematuria is also known as thin basement membrane disease (TBMD) and characterized by diffuse thinning of lamina densa of the glomerular basement membrane. We have performed the genetic linkage analysis between type IV collagen alpha 3 or alpha 4 chain and TBMD.But there was no linkage, suggesting the genes responsible for TBMD were other than these basement membrane collagens.2) Transcriptional factors in rat experimental glomerulonephritisWe have focused on protooncogenes such as c-fos, c-jun and c-myc, and AP-1 which is heterodimer complex of … More c-Fos and c-Jun, in rat anti Thy 1,1 nephritis and streptozotocin diabetic rats.mRNA expression for c-myc was maximal at 4 or 5 days after the induction anti Thy 1,1 nephritis, which phase is cosistent with mesangial cell proliferation. AP-1 complex expressed srtrongly in the chronic, glomeruloscrelotic phase of STZ diabetic rats rather than just after the induction of the diabetes. These results reveald that these transcriptional factors play an important roles in mesangial proliferative glomerulonephritis, especially in the progressive and chronic phase of the disease.3) Effects of all trans retinoic acid (ATRA) on mesangial cellsATRA is known as an inducer of cell differentiation. This may suggests that ATRA induces growth suppression of mesangial cells via the induction of phenotypic change of the cells.We have analyzed the effects of ATRA on cultured mesangial cells. ATRA suppressed the mesangial cell growth and mRNA expressions for c-fos, c-jun and c-myc. Then we administerd ATRA torats of anti Thy 1,1 nephritis. Mesangial cell number and matix score were significantly suppressed in ATRA treated rats. These results strongly suggested that the down regulation of these transcriptional factors could be a possible candidate for the treatment of mesangial proliferative glomerulonephritis. Less