Apoptosis and its related genes in renal epithelial cells of the stone-forming rat
Apoptosis and its related genes in renal epithelial cells of the stone-forming rat
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DOI：
10.1007/s00240-004-0434-6
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发表时间：
2005-02
影响因子：
--
通讯作者：
K. Miyazawa;Koji Suzuki;R. Ikeda;M. Moriyama;Y. Ueda;S. Katsuda
中科院分区：
文献类型：
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作者：
K. Miyazawa;Koji Suzuki;R. Ikeda;M. Moriyama;Y. Ueda;S. Katsuda
实验性高尿酸和草酸钙（CaOx）晶体与肾上皮损伤和细胞死亡有关。最近的一项研究表明，在体外的细胞凋亡中，尿酸盐诱导的增加，并推测这种现象可能有助于结石的形成。本研究观察了成石大鼠肾脏中凋亡细胞的发生率及凋亡相关基因的表达。雄性Wistar大鼠饮用含乙二醇的水，并灌胃1 α-OH-D3。在琼脂糖凝胶电泳中，细胞凋亡呈梯状。采用末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记法（TUNEL）对凋亡细胞进行定位。采用逆转录聚合酶链反应（RT-PCR）和免疫组化方法分析乳腺癌相关基因的表达。在对照组或给药第1天，通过TUNEL法未检测到标记，但在第3天开始在外髓质的肾小管上皮细胞中检测到标记，并且标记细胞的数量在观察期间进行性增加。2周后，大鼠肾脏中出现DNA片段的梯状条带。免疫组化显示Fas配体（Fas L）、Bax和白细胞介素1 β转换酶（ICE）在Henle袢降支和远曲小管上皮表达。ICE、c-myc、p53和Fas L基因的mRNA在结石大鼠肾脏中也有上调。
Experimental hyperoxaluria and calcium oxalate (CaOx) crystals are associated with renal epithelial injury and cell death. A recent study has demonstrated an oxalate-induced increase in cellular apoptosis in vitro, and speculates that this phenomenon may contribute to stone formation. We investigated the incidence of apoptotic cells and the expression of apoptosis related genes in the kidneys of stone-forming rats. Male Wistar rats were administrated ethylene glycol in drinking water and force fed with1α-OH-D3. Apoptosis was detected as a ladder of fragmented DNA in agarose gels of electrophoresed genomic DNA. Apoptotic cells were localized by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. The expression of apoptosis-related genes was analyzed by both reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. While no labeling was detected in the controls or on the first day of administration by the TUNEL method, labeling began to be detected in the renal tubular epithelium of the outer medulla at day 3, and the number of labeled cells increased progressively during the observation period. A ladder of DNA fragments was demonstrated in the kidneys of rats after 2 weeks. Immunohistochemical studies revealed the expression of Fas ligand (Fas L), Bax and interleukin-1 β converting enzyme (ICE) in the renal tubular epithelium of the descending limb of loop of Henle and the distal convoluted tubules. mRNA of the ICE, c-myc,p53and Fas L genes was also upregulated in the kidneys of the stone-forming rats.