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) 是一种有效的金属蛋白酶，可以降解各种类型的细胞外基质 (ECM)，如胶原蛋白 IV、V 和层粘连蛋白。有人认为，糖尿病引起的 ECM 积累至少部分是由于 MMP-2 的下调所致。然而，尚无MMP-2活性和表达是否影响糖尿病引起的体内肾损伤的数据。因此，我们研究了MMP-2缺乏对糖尿病动物肾小管间质损伤的影响。 材料和方法：用链脲佐菌素(STZ)(50mg/kg)诱导雄性MMP-2敲除小鼠(MMP-2 KO)和C57BL/6J小鼠(Ctrl)患糖尿病。 16周后，分别通过酶谱法和实时PCR测量皮质MMP-2表达和活性。通过酶联免疫吸附测定（ELISA）测量尿白蛋白排泄（UAE）和N-乙酰基-β-D-氨基葡萄糖苷酶（NAG）。通过蛋白质印迹和免疫组织化学评估阿尔法平滑肌肌动蛋白 (α-SMA)。分别通过苏木精和伊红染色 (HE) 和马森三色染色评估肾小管间质损伤和纤维化。 结果：与非糖尿病 Ctrl 小鼠相比，16 周糖尿病小鼠的血浆葡萄糖和 HbA1c 水平增加约 2-3 倍（血浆葡萄糖；490.3+/-7.5mg/dl， 糖化血红蛋白 ; 16 周糖尿病小鼠中为 9.91+/-0.22%）。酶谱分析和 RT-PCR 分析显示，糖尿病小鼠总肾皮质中 MMP-2 的表达和活性均增加。与 Ctrl DM 小鼠相比，MMP-2 KO DM 小鼠的 BUN、肌酐 (Cr) 和 UAE 血清水平显着升高（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 对比 0.16+/-0.03mg/mgCr，p<0.05）。此外，DM 小鼠中肾小管间质损伤和皮质 α-SMA 表达增强，而 MMP-2 KO DM 小鼠中肾小管间质损伤和皮质 α-SMA 表达进一步增加。结论：我们首次证明，尽管糖尿病小鼠中 MMP-2 水平增加，但糖尿病 MMP-2 敲除小鼠中肾小管间质损伤和纤维化加剧。本研究提示其他基质降解酶的减少可能与糖尿病肾病的肾小管间质损伤和纤维化有关。较少的

项目成果

Possible involvument 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: 2007
• 期刊: Journal of the American Society of Nephrology 18
• 作者: Kei Fukami;Sho-ichi Yamagishi;Seiji Ueda;Hiroshi Kawachi;Masavoshi Takeuchi;Seiya Okuda
• 通讯作者: Seiya Okuda

Circulating Matrix Metalloproteinase-2 Is an Independent Correlate of Proteinuria in Patients with Chronic Kidney Disease

• DOI: 10.1159/000151439
• 发表时间: 2009-01-01
• 期刊: AMERICAN JOURNAL OF NEPHROLOGY
• 影响因子: 4.2
• 作者: Nagano, Makio;Fukami, Kei;Okuda, Seiya
• 通讯作者: Okuda, Seiya

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

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