A Study for Elucidation of Functional Mechanism of High Molecular Weight Substances in Urine on Calcium Oxalate Stone Formation

阐明尿液中高分子物质对草酸钙结石形成作用机制的研究

基本信息

批准号：
12307032
负责人：
YACHIKU Sunao
金额：
$ 25.45万
依托单位：
Asahikawa Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

1. Study of crystal-cell interactions: An experimental model of calcium oxalate (CaOx) stones, suitable for investigation of crystal-cell interactions, has been established. It was confirmed that the phosphatidylserine exposure on the outer leaflet of the cell membrane of renal collecting duct cells could be evaluated quantitatively by using this model. Increased phosphatidylserine expressing cells and activation of the caspase family (intracellular signal transduction systems for apoptosis) were noted with increasing urinary oxalate excretion and calcium oxalate crystals. 2. Purification of stone-related proteins and their molecular biological analysis: Osteopontin (OPN) and calprotectin were purified. The adhesion of CaOx crystals to MDCK cells was promoted by OPN, while it was suppressed by thrombin and antisense oligonucleotide. It seems likely that OPN-DNA polymorphism in patients with urinary tract stones is involved in mutation of the enhancer regions that are related to synthes … More is of OPN protein. 3. OPN antisense gene transfer: The introduction of OPN antisense gene to renal tubular cells suppressed OPN expression and CaOx stone formation in the kidneys. The addition of OPN and vitronectin to renal tubular cells in culture elevated the intracellular expression of OPN and promoted the phosphorylation of focal adhesion kinase. Catechin suppressed the formation of CaOx crystals through inhibiting apoptosis induced by oxidative stress. 4. Study of urinary tract stone inhibitors using antisense gene: Human syndecan-1-expressing renal tubular cells were established. Heparan sulfate (HS) and HS proteoglycan were found to suppress the cellular injury by oxalate exposure and the adhesion of CaOx crystals to the cell surface. It was suggested that HS prevented the cell injury by serving as a charge barrier that inhibited the influx of oxalate ions into the cells. 5. Effects to stone formation by controlling the expression of stone-related protein mRNA: Increased expression of renal prothrombin F-1 (RPTF-1) mRNA in rat kidneys with CaOx was confirmed. In the macro-array analysis, it was noted that RPTF-1 protein, OPN gene, and the multiple genes encoding inflammatory systems were increased after CaOx crystal exposure to the renal tubular cells. Less

1。晶体 - 细胞相互作用的研究：已经建立了适合晶体 - 细胞相互作用的投资的草酸钙（CAOX）结石的实验模型。可以通过使用该模型来定量评估肾脏收集管细胞细胞膜外叶的磷脂酰丝氨酸暴露。增加的磷脂酰丝氨酸表达细胞和caspase家族的激活（2. 2的细胞内信号转导系统。与石材相关蛋白质的纯化及其分子生物学分析：骨桥蛋白（OPN）（OPN）和钙骨蛋白纯化，并通过OPN抑制了Caox Crysil的粘附于Opn和protsn。尿道结石患者的OPN-DNA多态性可能与合成的增强子区域的突变有关……更多的是OPN蛋白。 OPN的细胞内表达并促进了局灶性粘合剂激酶的磷酸化。 4。使用反义基因研究尿路抑制剂：人类Syndecan-1表达肾结通细胞。发现硫酸肝素（HS）和HS蛋白聚糖可通过草酸盐暴露和CAOX晶体粘附在细胞表面，从而抑制细胞损伤。有人提出，HS通过用作抑制草酸根离子对细胞影响的电荷屏障来防止细胞损伤。 5。通过控制结石相关蛋白mRNA的表达对石材形成的影响：用CAOX大鼠肾脏中肾脏蛋白F-1（RPTF-1）mRNA的表达增加。在宏观阵列分析中，注意到RPTF-1蛋白，OPN基因和编码炎症系统的多个基因在CAOX晶体暴露于肾小管细胞后增加。较少的