Molecular analysis of glucose transporter in glomerular mesangial cells

肾小球系膜细胞葡萄糖转运蛋白的分子分析

• 批准号: 03671143
• 负责人: KIKKAWA Ryuichi
• 金额: $ 0.32万
• 依托单位: Shiga University of Medical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03671143/
• 关键词: Diabetic nephropathy; Mesangial cells; Glucose transporter; IGF-I; Immunoblotting; Northern blot analysis; Immunoblotting

It has been known that the abnormalities of mesangial area in glomeruli play a crucial role in the progression of diabetic nephropathy, and the mesangial cells might participate in the evolution of these abnormalities. In order to clarify the role of mesangial cells, the cells were investigated under high concentration of glucose in vitro and various metabolic derangements have been detected, which might be related to the mesangial abnormalities seen in vivo. The glucose appears to cause various.metabolic abnormalities in mesangial cells after transported intracellularly via specific glucose transporter(GLUT). In this study, the glucose transporter in mesangial cells was characterized by western blot and northern blot analyses. The western blotting using monoclonal antibodies against GLUTs 1 and 4 showed the presence of GLUT 1 but failed to detect GLUT 4. The northern blot analysis using cDNAs for GLUT 1,2,3,4,and 5 revealed the presence of GLUT 1 mRNA only, which was compatible with the results of western blotting. Furthermore, insulin-like growth factor 1 (IGF-1), which was able to increase the glucose uptake in mesangial cells, was found to stimulate the gene expression of GLUT 1. Since the mesangial cells possesses an abundant amount of receptors for IGF-1 and the IGF-1 levels in renal tissue is increased in diabetic state, the amount of glucose transport in mesangial cells may be regulated by IGF-1 and be increased in diabetes mellitus. In summary, GLUT 1 is a main glucose transporter in mesangial cells, regulated by IGF-1, and may play a role in the evolution of metabolic derangements found in diabetes mellitus.

肾小球系膜区的异常在糖尿病肾病的发生、发展中起着重要作用，系膜细胞可能参与了这些异常的发生、发展过程。为了阐明系膜细胞的作用，在体外高浓度葡萄糖下研究了细胞，并检测到各种代谢紊乱，这可能与体内观察到的系膜异常有关。葡萄糖经细胞内特异性葡萄糖转运蛋白（GLUT）转运后，可引起系膜细胞多种代谢异常。在这项研究中，系膜细胞的葡萄糖转运蛋白的特点是通过蛋白质印迹和北方印迹分析。使用抗GLUT 1和4的单克隆抗体的蛋白质印迹法显示存在GLUT 1，但未能检测到GLUT 4。使用GLUT 1、2、3、4和5的cDNA进行的北方印迹分析显示仅存在GLUT 1 mRNA，这与蛋白质印迹的结果一致。此外，胰岛素样生长因子1（IGF-1），这是能够增加肾小球系膜细胞中的葡萄糖摄取，被发现刺激GLUT 1的基因表达。由于系膜细胞具有大量的IGF-1受体，并且在糖尿病状态下肾组织中的IGF-1水平增加，所以系膜细胞中的葡萄糖转运量可能受IGF-1调节，并且在糖尿病中增加。总之，GLUT 1是系膜细胞中的主要葡萄糖转运蛋白，受IGF-1调节，并且可能在糖尿病中发现的代谢紊乱的演变中发挥作用。

项目成果

梶原 信之,吉川 隆一,羽田 勝計,古家 大祐,戸川 雅樹,堀出 直樹,宇津 貴,杉本 俊郎,繁田 幸: "培養ラットメサンギウム細胞のブドウ糖輸送体機構の研究" 日本腎臓学会誌. 33. 1287 (1991)

Nobuyuki Kajiwara、Ryuichi Yoshikawa、Katsuke Hada、Daisuke Furuya、Masaki Tokawa、Naoki Horide、Takashi Utsu、Toshiro Sugimoto、Yuki Shigeta：“培养大鼠系膜细胞中葡萄糖转运机制的研究”日本肾脏病学会杂志 33. 1287。 (1991)

Masakazu Haneda, Ryuichi Kikkawa, Masaki Togawa, Daisuke Koya, Nobuyuki Kajiwara, Yukio Shigeta.: "Metabolic actions of insulin-like growth factor 1 in cultured glomerular mesangial cells." Metabolism. 40. 1311-1316 (1991)

Masakazu Haneda、Ryuichi Kikkawa、Masaki Tokawa、Daisuke Koya、Nobuyuki Kajiwara、Yukio Shigeta.：“胰岛素样生长因子 1 在培养的肾小球系膜细胞中的代谢作用。”

R.Kikkawa,N.Kajiwara,M.Haneda,Y.Shigeta,K.Tsuda,H.Yano,Y.Seino: "GLUT 1 is a main glucose transporter in rat mesangial cells" J Am Soc Nephrol. 3. 830 (1992)

R.Kikkawa、N.Kajiwara、M.Haneda、Y.Shigeta、K.Tsuda、H.Yano、Y.Seino：“GLUT 1 是大鼠系膜细胞中的主要葡萄糖转运蛋白”J Am Soc Nephrol。

M.Haneda,R.Kikkawa,M.Togawa,D.Koya,N.Kajiwara,Y.Shigeta: "Metabolic of insulin-like growth factor 1 in cultured glomerular mesangial cells" Metabolism. 40. 1311-1316 (1991)

M.Haneda，R.Kikkawa，M.Tokawa，D.Koya，N.Kajiwara，Y.Shigeta：“培养的肾小球系膜细胞中胰岛素样生长因子1的代谢”代谢。

前田 士郎,吉川 隆一,羽田 勝計 杉本 俊郎,古家 大祐,梶原 信之 宇津 貴,繁田 幸男: "ラット培養メサンギウム細胞における糖輸送担体(GLUTI)の調節構構" 日本腎臓学会誌.

Shiro Maeda、Ryuichi Yoshikawa、Katsuke Hada、Toshiro Sugimoto、Daisuke Furuya、Nobuyuki Kajiwara、Takashi Utsu、Yukio Shigeta：“培养大鼠系膜细胞中葡萄糖转运载体（GLUTI）的调节结构”日本肾病学会杂志。