Successful formation of calcium oxalate crystal deposition in mouse kidney by intraabdominal glyoxylate injection
Successful formation of calcium oxalate crystal deposition in mouse kidney by intraabdominal glyoxylate injection
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DOI：
10.1007/s00240-007-0082-8
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发表时间：
2007-04-01
影响因子：
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通讯作者：
Kohri, Kenjiro
中科院分区：
文献类型：
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作者：
Okada, Atsushi;Nomura, Shintaro;Kohri, Kenjiro
实验动物模型的建立将有助于研究肾结石的形成机制。目前已采用乙二醇法诱导大鼠钙性肾结石模型进行研究，但为了探讨影响肾结石形成的遗传学基础，从而有助于预防医学的发展，建立小鼠肾结石模型是必不可少的。本研究提出了在正常小鼠肾内诱导草酸钙结石的最佳条件。作为初步研究，不同剂量的草酸前体乙二醇酯、乙醇酸和乙二醇酯分别以自由饮用或腹腔注射的方式给药2个月。用光学显微镜、偏振光光学显微镜和电子显微镜检测结石的形成。采用X-射线衍射仪对石材成分进行检测。用免疫组织化学染色、原位杂交和定量逆转录聚合酶链式反应检测结石相关蛋白骨桥蛋白(OPN)的表达。即使在最高剂量的LD50下，乙二醇组或乙醇酸组也没有检测到肾结石。而乙醛(>60 mg/kg)组小鼠在给药后第3、6、9天发现大量肾结石。12天时，肾结石数量逐渐减少，15天时几乎检测不到结石。结石成分进一步分析为一水草酸钙。给予乙醛后，骨桥蛋白的表达显著增加。本研究建立了小鼠肾结石实验系统。诱发肾结石的难度表明，与大鼠相比，小鼠在高草酸应激下具有更强的内在防止结石形成的能力。小鼠和大鼠对高草酸应激的不同反应可能有助于肾结石形成的分子机制，并将有助于未来的预防医学。
The establishment of an experimental animal model would be useful to study the mechanism of kidney stone formation. A calcium kidney stone model in rats induced by ethylene glycol has been used for research; however, to investigate the genetic basis affecting kidney stone formation, which will contribute to preventive medicine, the establishment of a kidney stone model in mice is essential. This study indicates the optimum conditions for inducing calcium oxalate stones in normal mouse kidney. Various doses of oxalate precursors, ethylene glycol, glycolate and glyoxylate, were administered either by free drinking or intraabdominal injection for 2 months as a preliminary study. Stone formation was detected with light microscopy, polarized light optical microscopy and electron microscopy. Stone components were detected with X-ray diffraction analysis. The expression of osteopontin (OPN), a major stone-related protein, was detected with immunohistochemical staining, in situ hybridization and quantitative reverse transcriptase polymerase chain reaction. Kidney stones were not detected in ethylene glycol- or glycolate-treated groups even at the highest dose of LD50. Whereas, numerous kidney stones were detected in glyoxylate-treated mice (more than 60 mg/kg) at 3, 6 and 9 days after glyoxylate were administered intraabdominally. However, the number of kidney stones decreased gradually at day 12, and was hardly detected at day 15. The stone component was further analyzed as calcium oxalate monohydrate. A dramatic increase in the expression of OPN was observed by the administration of glyoxylate. We established a mouse kidney stone experimental system in this study. The difficulty of inducing kidney stones suggested that mice have greater intrinsic ability to prevent stone formation with hyperoxaluric stress than rats. The differing response to hyperoxaluric stress between mice and rats possibly contributes to the molecular mechanism of kidney stone formation and will aid preventive medicine in the future.