Gene expression and molecular mechanism of Annexins on urolithiasis

膜联蛋白在尿石症中的基因表达及分子机制

• 批准号: 16591627
• 负责人: MIYAZAWA Katsuhito
• 金额: $ 2.37万
• 依托单位: Kanazawa Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591627/
• 关键词: urolithiasis; calcium oxalate; calcium binding molecule; annexins; PAR; 蓚酸カルシウム; カルシウム結合蛋白遺伝子群; PARs; レーザーマイクロダイセクション

Purpose : Kidney stone formation is a complex process, and numerous molecules participate in this cascade. Attachment of newly formed crystals to renal epithelial cells appears to be a critical step in the development of this cascade. Annexins are one of the calcium binding molecules and up-regulated by calcium oxalate crystals. Annexin V is routinely used as a probe to demonstrate phosphatidylserine exposed on the surface of apoptotic cells. To obtain more detailed insight into annexins on urolithiasis and renal epithelial injury, we examined an incidence of apoptotic cells and expression of apoptosis related genes in kidneys of stone-forming rat.Materials and Methods : Male Wistar rats were administrated ethylene glycol (EG) in drinking water and force fed withl alpha-OH-D_3. 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 labe … More ling (TUNEL) method. Expression of apoptosis-related genes was analyzed by both reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry.Results : While no labeling was detected in the control or 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 administrated after 2 weeks. Immunohistochemical studies revealed the expressions of Fas ligand (Fas L), bax and interleukin-1 beta converting enzyme (ICE) in the renal tubular epithelium of the descending limb of loop of Henle and distal convoluted tubules. mRNA of ICE, c-myc, p53 and Fas L genes were also up-regulated in the kidneys of stone forming-rat model.Conclusion : In the hyperoxaluric, subsequent stone-forming rat kidney caused by the administration of EG and 1 alpha-OH-D_3, apoptosis was detected in the renal epithelial cells, especially in those of the descending limb of the loop of Henle and distal convoluted tubules. RT-PCR and immunohistochemical analyses revealed up-regulations of Fas-L, ICE, Bax, c-myc and p53. These findings indicate that renal epithelial injury in the stone-forming rat model following EG and 1 alpha-OH-D_3 feeding may induce the expression of those apoptosis-related genes, and different pathways in the stone-forming rat model may induce apoptosis. Less

目的：肾结石的形成是一个复杂的过程，许多分子参与了该级联。新形成的晶体附着在肾上皮细胞上似乎是该级联反应的关键步骤。膜联蛋白V通常用作探针，以证明在凋亡细胞表面暴露的磷脂酰丝氨酸。为了获得有关尿石症和肾上皮损伤的融合蛋白的更详细的见解，我们检查了凋亡细胞的事件，并在结石大鼠的肾脏中的凋亡相关基因的表达。材料和方法：雄性Wistar大鼠在饮用水中施用了乙烯甘氨酸（EG）的雄性Wistar大鼠（例如）在饮用水和抗水allpha-harpha-hapa-1-d_-d_-harpha-13中。在电泳基因组DNA的琼脂糖凝胶中，检测到凋亡是碎片DNA的梯子。凋亡细胞通过末端脱氧核苷酸转移介导的dutp-biotin nick End标签进行定位……更多的ling（TUNEL）方法。通过逆转录聚合酶链反应（RT-PCR）和免疫组织化学分析了与凋亡相关基因的表达：虽然在对照组中或通过TUNEL方法的管理中未检测到标记，但在肾小管上的肾小管上表皮中，在肾小管上的肾小球上均开始逐渐发现，在肾小管上的肾小球上，在肾小球上均开始进行标记。两周后，在大鼠行政的肾脏中证明了DNA碎片的梯子。免疫组织化学研究揭示了FAS配体（FAS L），BAX和白介素-1ββ（ICE）在Henle和远端备纤尿管下降的肾小管上皮的肾小管上皮中转化酶（ICE）的表达。冰，c-myc，p53和fas l基因的mRNA也被上调，在石头形成鼠标的孩子中。结论：在高氧钙钙疗法的，随后的石材形成大鼠肾脏中，由Eg的给药和1个alpha-oh-oh-d_3引起的，在肾上皮细胞中，尤其是在肾上皮细胞中检测到的，尤其是在那些renal上皮细胞中，属于这些疾病的范围，这些疾病是在这些范围内的;小管。 RT-PCR和免疫组织化学分析揭示了FAS-L，ICE，BAX，C-MYC和P53的上调。这些发现表明，例如和1个α-OH-D_3喂养后，石材形成大鼠模型中的肾上皮损伤可能诱导这些与凋亡相关的基因的表达，而石材形成大鼠模型中的不同途径可能诱导凋亡。较少的

项目成果

腎尿細管上皮細胞におけるProtease-activated receptorの発現

肾小管上皮细胞中蛋白酶激活受体的表达

• 发表时间: 2005
• 期刊: 日尿結石誌 4・2
• 作者: Yamashita M;Kajiyama H;Kikkawa F;Oomori M;宮澤 克人
• 通讯作者: 宮澤 克人

Apoptosis and its related genes in renal epithelial cells of the stone-forming rat

• DOI: 10.1007/s00240-004-0434-6
• 发表时间: 2005-02
• 期刊: Urological Research
• 作者: K. Miyazawa;Koji Suzuki;R. Ikeda;M. Moriyama;Y. Ueda;S. Katsuda

尿路結石の再発予防法

如何预防尿路结石复发

• 发表时间: 2006
• 期刊: 総合臨床 55-3
• 作者: Grover PK;Miyazawa K et al.;Haraguchi CM;Shoda T;宮澤 克人 他
• 通讯作者: 宮澤 克人 他

Cyclooxgenase 2およびプロスタグランジンE2の蓚酸カルシウム結晶付着の影響について

草酸钙晶体附着对环氧化酶 2 和前列腺素 E2 的影响

• 发表时间: 2004
• 期刊: 日尿結石誌 3・2
• 作者: 宮澤 克人;森山 学;鈴木 孝治
• 通讯作者: 鈴木 孝治

The preventive treatment of patients with urinary stone disease.

泌尿系结石病患者的预防性治疗。

• 发表时间: 2006
• 期刊: Sogorinsyo 55
• 作者: 鈴木孝太;笠井 剛;藤江道子;エンフマダバサンブー;鈴木真梨子;島 崇;島津由加里;和田麻美子;三宅麻喜;正田朋子;永井聖一郎;平田修司;星 和彦;Kurioka Hiroko;Shoda T;KATSUHITO MIYAZAWA;Wada M;Nagai S;Goto Sakae;Takuya Mitsui;KATSUHITO MIYAZAWA
• 通讯作者: KATSUHITO MIYAZAWA