Tubulointerstitial injury is exacerbated in streptozotocin-induced diabetic matrix metalloproteinase-2 knockout mice

链脲佐菌素诱导的糖尿病基质金属蛋白酶-2敲除小鼠肾小管间质损伤加剧

• 批准号: 19590970
• 负责人: FUKAMI Kei
• 金额: $ 2.75万
• 依托单位: Kurume University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19590970/
• 关键词: 糖尿病性腎症; 細胞外基質; MMP-2; 尿細管間質; 尿蛋白; Diabetic Nephropathy; AGE; podocyte

Background and Aims : Matrix metalloproteinnase-2 (MMP-2) is one of the potent metalloproteinases which can degrade various types of extracellular matrix (ECM) such as collagen IV, V and laminin. It has been suggested that diabetes-induced accumulation of ECM are, at least in part, by down-regulation of MMP-2. However there is no data whether MMP-2 activity and expression affect diabetes-induced renal damage in vivo. Therefore, we investigated the effects of MMP-2 deficiency on tubulointerstitial injury in diabetic animals.Materials and Methods : Diabetes was induced by streptozotocin (STZ)(50mg/kg) in male MMP-2 knockout mice (MMP-2 KO) and C57BL/6J mice (Ctrl). After 16 weeks, cortical MMP-2 expression and activity were measured by zymography and real-time PCR, respectively. Urinary albumin excretion (UAE) and N-acetyl-β-D-glucosaminidase (NAG) were measured by enzyme-linked immunosorbent assay (ELISA). Alfa-smooth muscle actin (α-SMA) was evaluated by western blots and immnohistoche … More mistry. Tubulointerstitial injury and fibrosis were evaluated by hematoxylin and eosin staining (HE) and Masson-trichrome staining, respectively.Results : Plasma levels of glucose and HbA1c were increased by about 2-3-folds in 16-week diabetic mice compared with non-diabetic Ctrl mice (plasma glucose ; 490.3+/-7.5mg/dl, HbA1c ; 9.91+/-0.22% in 16-week diabetic mice). MMP-2 expression and activity in the total kidney cortex of diabetic mice were increased in zymography and RT-PCR analysis. Serum levels of BUN, creatinine (Cr) and UAE were significantly increased in MMP-2 KO DM mice compared with Ctrl DM mice (BUN ; 23.2+/-1.7 vs 39.8+/-6.0mg/dl, p<0.01, Cr ; 0.07+/-0.01 vs 0.17+/-0.04mg/dl, p<0.01, UAE ; 0.08+/-0.01 vs 0.16+/-0.03mg/mgCr, p<0.05). Further, tubulointerstitial injury and cortical α-SMA expression were enhanced in DM mice, which were further increased in MMP-2 KO DM mice.Conclusions : We demonstrate for the first time that tubulointerstitial injury and fibrosis were exacerbated in diabetic MMP-2 knockout mice, even though MMP-2 levels were increased in diabetic mice. The present study suggests that the decrease in other matrix-degrading enzymes may be involved in the tubulointerstitial injury and fibrosis in diabetic nephropathy. Less

背景和目的：基质金属蛋白酶-2（MMP-2）是潜在的金属蛋白酶之一，可以降解各种类型的细胞外基质（ECM），例如胶原蛋白IV，V，V和椎板蛋白。已经提出，糖尿病诱导的ECM的积累至少部分通过下调MMP-2。但是，没有数据是否会影响糖尿病诱导的体内肾脏损伤。因此，我们研究了MMP-2缺乏对糖尿病动物中肾小管间质损伤的影响。材料和方法：糖尿病是由链蛋白酶（STZ）（STZ）（50mg/kg）诱导的雄性MMP-2基因敲除小鼠（MMP-2 KO）和C57BL/6J小鼠（C57BL）。 16周后，分别通过Zymography和实时PCR测量皮质MMP-2表达和活性。尿白蛋白排泄（UAE）和N-乙酰基-β-D-葡萄糖酰胺酶（NAG）通过酶联免疫吸附测定法（ELISA）测量。通过Western印迹和IMMNO-HERTOCHE评估Alfa-Smooth肌肉肌动蛋白（α-SMA）……更糟糕。分别通过苏木精和曙红染色（HE）和群质染色评估了肾小管间质损伤和纤维化，分别是16周糖尿病小鼠的血浆葡萄糖和HBA1C的血浆水平增加了约2-3倍，与非糖尿病ctrl小鼠相比; plasma glucose; plasma glucose; 490+/dn90; 490.490 l.490+/d。 HBA1C; 16周糖尿病小鼠中的9.91 +/- 0.22％。在Zymography和RT-PCR分析中，糖尿病小鼠总肾脏皮层中的MMP-2表达和活性增加。 Serum levels of BUN, creatinine (Cr) and UAE were significantly increased in MMP-2 KO DM mice compared with Ctrl DM mice (BUN; 23.2+/-1.7 vs 39.8+/-6.0mg/dl, p<0.01, Cr; 0.07+/-0.01 vs 0.17+/-0.04mg/dl, p<0.01, UAE; 0.08+/-0.01 vs 0.16 +/- 0.03mg/mgcr，p <0.05）。此外，在MMP-2 KO DM小鼠中，DM小鼠中的微管区间损伤和皮质α-SMA表达增强了。结论：我们首次证明，在糖尿病MMP-2基因敲除小鼠中，MMP-2级别在糖尿病中增强了糖尿病的损伤和纤维化的损伤和纤维化。本研究表明，其他基质降解酶的降低可能与糖尿病性肾病中的肾上头损伤和纤维化有关。较少的

项目成果

Administration of pigment epithelium-derived factor (PEDF) reduces proteinuria by suppressing decreased nephrin and increased VEGF expression in the glomeruli of adriamycin-iniected rats.

给予阿霉素感染大鼠肾小球中色素上皮衍生因子（PEDF）可通过抑制去氧肾上腺素减少和血管内皮生长因子表达增加来减少蛋白尿。

• 发表时间: 2008
• 期刊: Nephrol Dial Transplant. 24
• 作者: Fujimura T;Fukami K
• 通讯作者: Fukami K

Olmesartan blocks advanced glycation end products (AGEs)-induced angiogenesis in vitro by suppressing receptor for AGES (RAGE) expression

• DOI: 10.1016/j.mvr.2007.05.001
• 发表时间: 2008-01-01
• 期刊: MICROVASCULAR RESEARCH
• 影响因子: 3.1
• 作者: Yamagishi, Sho-ichi;Matsui, Takanori;Imaizumi, Tsutomu
• 通讯作者: Imaizumi, Tsutomu

Inhibition of NADPH oxidase prevents advanced glycation end product-mediated damage in diabetic nephropathy through a protein kinase C-alpha-dependent pathway.

NADPH 氧化酶的抑制可通过蛋白激酶 C-α 依赖性途径预防糖尿病肾病中晚期糖基化终产物介导的损伤。

• 发表时间: 2008
• 期刊: Forbes JM. Diabetes. 57
• 作者: Thallas-Bonke V;Thorpe SR;Coughlan MT;Fukami K;Yap FY;Sourris KC;Penfold SA;Bach LA;Cooper ME
• 通讯作者: Cooper ME

Possible involvement of AGE and MMP-2 in podocyte detachment and loss.

AGE 和 MMP-2 可能参与足细胞脱离和丢失。

• 发表时间: 2008
• 作者: Kei Fukami;Sho-ichi Yamagishi;Seiji Ueda;Seiya Okuda;Fukami K
• 通讯作者: Fukami K

Development of Enzyme-Linked Immunosorbent Assay System for PEDF and its Clinical Utility

• DOI: 10.2174/156652410791065318
• 发表时间: 2010-04-01
• 期刊: CURRENT MOLECULAR MEDICINE
• 影响因子: 2.5
• 作者: Fukami, K.;Yamagishi, S. -I.;Okuda, S.
• 通讯作者: Okuda, S.