Role of SGLT3 in diabetes-mediated increased renal sodium reabsorption

SGLT3 在糖尿病介导的肾钠重吸收增加中的作用

• 批准号: 8282840
• 负责人: NILOOFAR M TABATABAI
• 金额: $ 24.98万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-15 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8282840
• 关键词: Affinity; Age; Agonist; Amino Acid Sequence; Amino Acid Sequence Homology; Animal Model; Animals; Antibodies; Apical; Binding; Blood Pressure; Brain; C57BL/6 Mouse; COS-7 Cell; Cadmium; Carrier Proteins; Cell Line; Cell membrane; Cells; Chronic; Colon; Confocal Microscopy; Custom; Data; Development; Diabetes Mellitus; Diabetic Nephropathy; Diabetic mouse; End stage renal failure; Epithelial Cells; Exposure to; Family; Family suidae; Fluorescence; Fluorescent Antibody Technique; Genes; Glucose; Glucose Transporter; Goals; Health; Human; Hyperglycemia; Hypertension; Immunoblotting; In Vitro; Intestines; Investigation; Ion Channel; Ions; Kidney; Knock-out; Knowledge; Malignant Epithelial Cell; Measures; Mediating; Mediator of activation protein; Membrane Protein Traffic; Messenger RNA; Microscopy; Modeling; Mouse Strains; Mus; Muscle Fibers; Na(+)-K(+)-Exchanging ATPase; Non-Insulin-Dependent Diabetes Mellitus; Oocytes; Ovary; Phlorhizin; Physiological Processes; Play; Property; Protein Isoforms; Proteins; Proximal Kidney Tubules; Rattus; Renal carcinoma; Research Design; Risk Factors; Role; Secondary to; Side; Small Intestines; Sodium; Sodium Chloride; Techniques; Testing; Time; Tissues; United States; Water; Xenopus laevis; Xenopus oocyte; absorption; base; design; diabetic; diabetic patient; glucose analog; glucose transport; glucose uptake; in vivo; kidney cell; knock-down; mouse model; non-diabetic; novel; public health relevance; response; small hairpin RNA; solute; sugar; uptake; young adult

DESCRIPTION (provided by applicant): Diabetic nephropathy is the leading cause of end-stage renal disease in the United States. Hyperglycemia has been recognized as a major risk factor in the development and progression of diabetic nephropathy. Hyperglycemia-induced increase in proximal tubule (PT) Na+ retention has been shown in diabetic patients and in animal models of diabetes. This enhanced Na+ reabsorption may play a role in the development of hypertension, which is an additional contributing factor in the development of diabetic nephropathy and the end stage renal disease. A role for a PT sodium-glucose cotransporter (SGLT) in hyperglycemia-induced increased sodium retention has been suggested but this transporter has not yet been identified. Kidney expresses SGLT1 and SGLT2 on the apical side of the PT cells. These proteins play crucial roles in sodium-dependent uptake of glucose from the glomerular filtrate. SGLT3 mRNA has been found in the human kidney carcinoma cells and in the pig kidney. When over-expressed in Xenopus oocytes, human SGLT3 did not transport glucose but in response to glucose, it mediated inward flux of sodium. Mouse has two genes encoding SGLT3a and 3b and we have shown that their mRNAs were expressed in the kidney and in the cultured kidney cells from mouse. We also showed while mRNA levels of SGLT3s in cultured mouse primary kidney cells exposed to cadmium were several folds higher than in their levels in untreated cells, the sodium-dependent uptake of glucose in Cd-treated cells had decreased supporting that SGLT3 is not a glucose transporter. Based on the above and additional preliminary studies with the potent agonist of SGLT3, deoxynojirimycin (DNJ), we hypothesize that kidney SGLT3 serves as a novel glucose-stimulated Na+ transporter in the PT that may play role in hyperglycemia-induced Na+ retention in diabetes. To test our hypothesis, we propose: (1) To localize the SGLT3 protein in human and mouse kidneys, (2) To determine the role of SGLT3 in glucose-mediated Na+ uptake in PT cells in vitro, (3) To determine the role of SGLT3 in hyperglycemia-mediated PT Na+ reabsorption in vivo. The proposed in vitro and in vivo studies are designed to investigate the role of SGLT3 as a novel glucose-stimulated Na+ transporter that may play role in the enhanced Na+ reabsorption in diabetes. PUBLIC HEALTH RELEVANCE: One of the major adverse health effects of diabetes is damage to the kidneys. Increased kidney salt reabsorption in diabetes may play a role in the development of high blood pressure, which in turn can cause damage to the kidneys. The mechanism of diabetes-induced increased salt reabsorption is not known. We have evidence that a novel sodium transporter may be a mediator. The goal of this study is to show that human kidneys express this protein and also to show that glucose stimulates sodium uptake by this transporter. This study is designed to identify the mechanism for the enhanced salt retention in diabetes.

描述(申请人提供)：糖尿病肾病是美国终末期肾病的主要原因。高血糖已被认为是糖尿病肾病发生发展的主要危险因素。在糖尿病患者和糖尿病动物模型中，高血糖诱导的近端小管(PT)钠离子滞留增加。这种增强的Na+重吸收可能在高血压的发展中起作用，高血压是糖尿病肾病和终末期肾脏疾病发展的另一个促成因素。PT钠-葡萄糖共转运体(SGLT)在高血糖诱导的钠滞留增加中的作用已被提出，但该转运体尚未被确定。肾脏在PT细胞顶侧表达SGLT1和SGLT2。这些蛋白在钠依赖的肾小球滤液葡萄糖摄取中起着关键作用。已在人肾癌细胞和猪肾中发现了SGLT3mRNA。当人SGLT3在非洲爪哇卵母细胞中过表达时，它不转运葡萄糖，但对葡萄糖的反应，它介导了钠的内流。小鼠有两个编码SGLT3a和3b的基因，我们已经证明它们的mRNAs在小鼠肾脏和培养的小鼠肾脏细胞中都有表达。我们还发现，虽然镉暴露培养的小鼠原代肾细胞SGLT3s的mRNA水平比未处理的细胞高出数倍，但镉处理细胞对葡萄糖的钠依赖摄取减少了，这支持了SGLT3不是葡萄糖转运体。基于上述研究和对SGLT3激动剂脱氧诺吉霉素(DNJ)的初步研究，我们推测肾脏SGLT3在糖尿病高血糖诱导的Na+滞留中可能是一种新的葡萄糖刺激的PT中的Na+转运体。为了验证我们的假设，我们建议：(1)定位SGLT3蛋白在人和小鼠肾脏中的位置；(2)确定SGLT3在体外葡萄糖介导的PT细胞Na+吸收中的作用；(3)确定SGLT3在体内高血糖介导的PT Na+重吸收中的作用。这项体外和体内研究旨在探讨SGLT3作为一种新的葡萄糖刺激的Na+转运体在糖尿病患者增强Na+重吸收中的作用。 公共卫生相关性：糖尿病对健康的主要不利影响之一是对肾脏的损害。糖尿病患者肾脏盐重吸收增加可能在高血压的发展中起作用，而高血压反过来又会对肾脏造成损害。糖尿病引起的盐重吸收增加的机制尚不清楚。我们有证据表明，一种新的钠转运体可能是一种介体。这项研究的目的是证明人类肾脏表达这种蛋白质，并证明葡萄糖通过这种转运蛋白刺激钠的摄取。这项研究旨在确定糖尿病患者食盐滞留增加的机制。