RESEACH FOR THE PATHOGENIC MECHANISM OF DIABETIC NEUROPATHY

糖尿病神经病发病机制的研究

• 批准号: 12671107
• 负责人: NAKAMURA Jiro
• 金额: $ 2.18万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671107/
• 关键词: Diabetic Neuropathy; Schwannoma Celis; Polyol Pahtway; Protein Kinase C; Aldosb Reduclase Inhibitior; プロティンキナーゼC

Aims : Polyol pathway hyperactivity and altered PKC activity have been proposed as the pathogenic mechanism of diabetic neuropathy. However, the relationship between polyol pathway and PKC activity has not been precisely investigated. The present study was conducted to investigate the effects of high glucose and polyol pathway on the PKC-MAPK cascade and cell growth using.cultured rat Schwannoma cells (JS-1 cells).Methods : JS-1 cells were cultured in 5.5 or 20 mM glucose (HG) with or without epalrestat (Ep ; 1μM) for 14 days, or treated with an antisense against PKC-α (AS) or a p38 MAPK specific inhibitor, SB-203580 (SB). The proliferation activity by assay of [^3H]-thyraidine uptake (% of control), PKCα activity by its protein expression in membrane fraction, and p38 activity by the ratio of phosphorylated to total p38 protein expression were measured.Results : 1) PKC-a and p38 activityies were decreased under the HG condition, which were ameliorated by Ep. 2) With AS treatment, the protein expression of PKC-α and p3S activity were decreased in a time- and dose-dependent fashion. 3) Proliferation activity was decreased by both AS and SB in a dose-dependent fashion. Conclusions: These results suggest that PKC-α and p38 MAPK activities are decreased by high glucose through the alclose reductase-sensit.ive pathway, and that PKC-α-p38 cascade would play an important- role in proliferation of neural cells, indicating,that glucose-induced polyol. Pathway hyperactivity would deteriorate the proI.ifernIion of Sdiwmm cells through' the decreased activities of PKC-α and p38 MAPK, leading t.o diabetic neuropathy.

目的：多元醇途径过度活跃和PKC活性改变被认为是糖尿病神经病变的发病机制。然而，多元醇途径与PKC活性之间的关系还没有得到准确的研究。以培养的大鼠神经鞘瘤细胞(JS-1细胞)为研究对象，观察高糖和多元醇途径对PKC-MAPK级联反应和细胞生长的影响。方法：将培养的JS-1细胞置于含或不含依帕司他(EP；1μM)的5.5mM或20 mM葡萄糖中培养14天，或用反义PKC-α(AS)或p38MAPK特异性抑制剂SB-203580(SB)处理。结果：1)在HG条件下，PKC-a和p38活性降低，EP可使HG条件下PKC-a和p38活性降低，而α活性则由膜蛋白表达水平决定。2)AS处理后，PKC-α蛋白表达和P3S活性均呈时间和剂量依赖性降低。3)As和Sb均呈剂量依赖性抑制细胞增殖活性。结论：高糖可通过ALCLO还原酶敏感性途径降低PKC-α和p38MAPK活性，PKC-α-p38级联通路在神经细胞增殖中可能起重要作用，提示葡萄糖诱导的神经细胞增殖是由葡萄糖诱导的。通路的过度活跃会通过PKC-α和p38MAPK活性的降低而恶化SDIwmm细胞的增殖，导致T.O糖尿病神经病变。

项目成果

Nakamura J.: "Glucose-induced hyperproliferation of cultured rat aortic smooth muscle cells through polyol pathway hyperactivity"Diabetologia. 44. 480-487 (2001)

Nakamura J.：“通过多元醇途径过度活跃，葡萄糖诱导培养的大鼠主动脉平滑肌细胞过度增殖”Diabetologia。

Yasuda Y: "Role of PKC and TGF-β receptor in glucose-induced proliferation of smooth muscle cells"Biochem Biophys Res Commun.. 281. 71-77 (2001)

安田 Y：“PKC 和 TGF-β 受体在葡萄糖诱导的平滑肌细胞增殖中的作用”Biochem Biophys Res Commun. 281. 71-77 (2001)

Jiro Nakamura: "Physiological and morphometric analysis of neuropathy in sucrose-fed OLETF rats"Elsevier Diabeles Research and Clinical Practice. 51. 9-20 (2001)

Jiro Nakamura：“蔗糖喂养的 OLETF 大鼠神经病变的生理和形态测量分析”Elsevier 糖尿病研究和临床实践。

Naruse K: "Aldose reductase inhibition prevents glucose-induced apoptosis in cultured bovine retinal microvascular pericytes"Exp Eye Res. 71. 309-315 (2000)

Naruse K：“醛糖还原酶抑制可防止培养的牛视网膜微血管周细胞中葡萄糖诱导的细胞凋亡”Exp Eye Res。

Nakamura J: "Glucose-induced hyperproliferation of cultured rat aortic smooth muscle cells through polyol pathway hyperactivity"Diabetologia. 44. 480-487 (2001)

Nakamura J：“通过多元醇途径过度活跃，葡萄糖诱导培养的大鼠主动脉平滑肌细胞过度增殖”Diabetologia。