The Detection Of Different Genes Expression From Renal Tissue Of ATSN Rats And The Effects Of PDTC On The Pathologic Changes Of Rats And The Effects Of PDTC On The Pathologic Changes Of Rats With ATSN

AIMS: (1) To reproduce rat model of mesangial proliferative glomerulonephritis (MPGN), namely, anti-thymocyte serum induced nephritis (ATSN), and to observe the pathologic changes of ATSN in different stages. (2) To detect expressive genes from renal tissue of rats with ATSN in apoptotic and protiferative phases and measure the mRNA level of two apoptosis-related genes (nerve growth factor induced protein I-B, NGFI-B and growth and DNA damage 45 gamma, Gadd45γ) for exploring the reason of glomerular mesangial cell (MC) apoptosis. (3) To study the expression of glomerular nuclear factor-KB ( NF-kB ) and effect of pyrrolidine dithiocarbamate (PDTC ) as the specific inhibitor of NF-kB on the pathologic changes of ATSN rats at different time. METHODS: (1) The ATSN model of rat was reproduced with anti-thymocyte serum(ATS). The pathologic changes such as MC apoptosis, lysis and proliferation were examined by light microscope(LM), electron microscope(EM), immunofluorescence(IF), immunohistochemical staining, and Tdt-mediated X-dUTP nick end labeling (Tunel) at experimental 40min, 24h and 7d after injecting ATS or normal serum. (2) The changes of gene expression profiles from renaltissue of rats with ATSN at 40min and 5d were detected by genechip technique, then, up-regulated genes were analyzed in Genbank in order to find some functions. The NGFI-B and Gadd45y were chosen and their mRNA level was measured by reverse transcription PCR (RT-PCR) and real-time PCR. (3) The 24 rats were divided into three groups, namely, ATSN group, ATSN+PDTC group and Control group. The expression of glomerular NF-kB p65 and the inhibitory effect of PDTC on pathologic changes of ATSN were examined by immunohistochemical staining, light microscope, electron microscope and Tunel at experimental 40min, 24h and 7d. RESULTS: (1) MC showed lysis, and glomerular cell numbers decreased at 24h, then increased in ATSN group at 7d under light microcope. In addition, the deposition of rabbit IgG and rat C3, the expression of PCNA and ct-SMA were seen in the glomeruli of rats with ATSN at different time. Under electron microcope, glomerular MC showed apoptosis at 40min, lysis at 24h, proliferation and extracellular matrix (ECM) accumulation at 7d. The apoptotic cell positive staining was increased in glomeruli by Tunel, and with the progess of disease, positive cells were decreased gradually. The mentioned-above results indicated that MC apoptosis, lysis and secondary proliferation were indeed appeared in ATSN model rats as the Shimizu's reports. (2) The 341 up-regulated expression genes (including apoptosis, proliferation and signal transduction related genes) and 392 down-regulated expression...