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，许多分子参与了这个级联反应。新形成的晶体附着在肾上皮细胞上似乎是这个级联发展的关键步骤。膜联蛋白是钙结合分子之一，受到草酸钙晶体的上调。膜联蛋白V通常被用作探针来显示暴露在凋亡细胞表面的磷脂酰丝氨酸。为了更详细地了解膜联蛋白对尿石症和肾上皮损伤的影响，我们检测了结石形成大鼠肾脏中凋亡细胞的发生率和凋亡相关基因的表达。材料与方法：雄性Wistar大鼠饮水中加入乙二醇（EG），灌喂α - oh - d_3。在电泳基因组DNA的琼脂糖凝胶中检测到细胞凋亡是DNA片段的阶梯。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（TUNEL）法定位凋亡细胞。采用逆转录聚合酶链反应（RT-PCR）和免疫组织化学分析凋亡相关基因的表达。结果：TUNEL法在对照组和给药第1天未检测到标记，第3天外髓质肾小管上皮开始检测到标记，观察期间标记细胞数量逐渐增加。给药2周后，大鼠肾脏出现DNA片段阶梯状排列。免疫组化结果显示，肾小管上皮内Fas配体（Fas L）、bax和白细胞介素-1 β转换酶（ICE）在Henle环降肢和远曲小管中表达。结石形成模型大鼠肾脏中ICE、c-myc、p53和Fas L基因mRNA表达也上调。结论：EG和1 α - oh - d_3致高血氧血症大鼠肾结石形成后，肾上皮细胞出现凋亡，尤以肾大袢降支和远曲小管细胞凋亡最明显。RT-PCR和免疫组化分析显示Fas-L、ICE、Bax、c-myc和p53表达上调。上述结果提示，EG和1 α - oh - d_3喂养后大鼠肾上皮损伤可能诱导这些凋亡相关基因的表达，并且在结石形成大鼠模型中不同的通路可能诱导细胞凋亡。少

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

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

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

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

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