Oseto Susumu, Moriyama Toshiki, Kawada Noritaka, Nagatoya Katsuyuki, Takeji Masanobu, Ando Akio, Yamamoto Tadashi, Imai Enyu, Hori Masatsugu
Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Osaka, Japan.
Kidney Int. 2003 Oct;64(4):1241-52. doi: 10.1046/j.1523-1755.2003.00219.x.
All-trans retinoic acid (ATRA) has antiproliferative and anti-inflammatory effects and is currently used in the treatment of leukemia and dermatologic diseases. We tested the therapeutic potential of ATRA on anti-glomerular basement membrane (GBM) glomerulonephritis rats.
Glomerulonephritis was induced in male Wistar-Kyoto rats on day 0 by an intravenous injection of antirat GBM antibody. On day 14 after the induction of anti-GBM glomerulonephritis, some rats were sacrificed (N = 5). Another 10 rats were divided into two groups: the vehicle group (N = 5) and the ATRA treated group (N = 5). ATRA was orally administrated from day 14 to day 27 after disease induction. Blood pressure, body weight, urinary protein excretion, and blood chemistry was determined on days 1, 14, 21, and 27. Kidney samples were obtained on day 28. The kidneys were examined with periodic acid-Schiff staining (PAS) and immunohistochemistry using antibodies against the proliferative cell nuclear antigen (PCNA), rat monocyte and macrophage (ED-1), and alpha-smooth muscle actin (alpha-SMA). Glomerular RNA was extracted from isolated glomeruli, and reverse transcription (RT) followed by polymerase chain reaction (PCR) was performed.
ATRA administration produced a 55% reduction of proteinuria in glomerulonephritis rats. Light microscopic analysis revealed severe necrosis/crescent formation (>50% of the glomerulus) affecting 34% of glomeruli in vehicle rats, whereas ATRA treatment reduced the glomeruli showing severe change to 14%. ATRA also significantly reduced PCNA-positive cells, ED-1-positive cells and alpha-SMA-positive area in the glomeruli. RT-PCR analyses revealed that a wide variety of genes including inflammation related [tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and CCAAT enhancer-binding protein delta (C/EBPdelta)], cell proliferation-related [platelet-derived growth factor (PDGF)] and fibrosis-related [transforming growth factor-beta1 (TGF-beta1), type I collagen, and alpha-SMA) genes were suppressed in the glomeruli of ATRA-treated rats.
ATRA administration significantly reduced severe necrosis/crescent formation and urinary protein excretion in glomerulonephritis rats. Suppression of a wide variety of gene expression may partly explain the mechanism of ATRA's antiproliferative and anti-inflammatory effects. These data suggest a novel therapeutic application of ATRA toward glomerulonephritis.
全反式维甲酸（ATRA）具有抗增殖和抗炎作用，目前用于治疗白血病和皮肤病。我们测试了ATRA对抗肾小球基底膜（GBM）肾小球肾炎大鼠的治疗潜力。
在第0天通过静脉注射抗大鼠GBM抗体，诱导雄性Wistar-Kyoto大鼠发生肾小球肾炎。在诱导抗GBM肾小球肾炎后的第14天，处死部分大鼠（N = 5）。另外10只大鼠分为两组：溶剂对照组（N = 5）和ATRA治疗组（N = 5）。从疾病诱导后的第14天至第27天口服给予ATRA。在第1、14、21和27天测定血压、体重、尿蛋白排泄和血液生化指标。在第28天获取肾脏样本。使用针对增殖细胞核抗原（PCNA）、大鼠单核细胞和巨噬细胞（ED-1）以及α-平滑肌肌动蛋白（α-SMA）的抗体，通过过碘酸-希夫染色（PAS）和免疫组织化学检查肾脏。从分离的肾小球中提取肾小球RNA，并进行逆转录（RT）随后进行聚合酶链反应（PCR）。
给予ATRA可使肾小球肾炎大鼠的蛋白尿减少55%。光学显微镜分析显示，溶剂对照组大鼠中34%的肾小球出现严重坏死/新月体形成（>50%的肾小球），而ATRA治疗使出现严重病变的肾小球减少至1·4%。ATRA还显著减少了肾小球中PCNA阳性细胞、ED-1阳性细胞和α-SMA阳性区域。RT-PCR分析显示，在ATRA治疗的大鼠肾小球中，包括炎症相关基因[肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和CCAAT增强子结合蛋白δ（C/EBPδ）]、细胞增殖相关基因[血小板衍生生长因子（PDGF）]以及纤维化相关基因[转化生长因子-β1（TGF-β1）、I型胶原和α-SMA]在内的多种基因表达受到抑制。
给予ATRA可显著减少肾小球肾炎大鼠的严重坏死/新月体形成和尿蛋白排泄。多种基因表达的抑制可能部分解释了ATRA抗增殖和抗炎作用的机制。这些数据提示ATRA在肾小球肾炎治疗中的新应用。
