Cellular dysfunction in the diabetic fibroblast - Impairment in migration, vascular endothelial growth factor production, and response to hypoxia
Cellular dysfunction in the diabetic fibroblast - Impairment in migration, vascular endothelial growth factor production, and response to hypoxia
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DOI：
10.1016/s0002-9440(10)63821-7
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发表时间：
2003-01-01
影响因子：
6
通讯作者：
Gurtner, GC
中科院分区：
文献类型：
--
作者：
Lerman, OZ;Galiano, RD;Gurtner, GC
虽然已知全身性疾病（如糖尿病）会导致伤口愈合受损，但这种损伤的机制尚不清楚。由于成纤维细胞是必不可少的伤口修复，我们比较了体外培养的成纤维细胞的行为糖尿病，瘦素受体缺陷（db/db）小鼠与野生型成纤维细胞从小鼠相同的遗传背景中的过程中重要的组织修复。与正常成纤维细胞相比，糖尿病小鼠成纤维细胞迁移率降低75%（P < 0.001），缺氧（1%O-2）对迁移率无明显刺激作用，而野生型成纤维细胞迁移率在缺氧条件下上调近2倍（P < 0.05）。明胶酶谱法和酶联免疫吸附试验均显示糖尿病成纤维细胞基质金属蛋白酶-9原的表达量是正常成纤维细胞的2倍（P < 0.05）。与野生型成纤维细胞相比，成年糖尿病成纤维细胞在血管内皮生长因子（VEGF）的产生方面表现出七倍的损害（4.5 +/-1.3pg/ml对34.8 +/-3.3pg/ml，P < 0.001）。此外，野生型成纤维细胞VEGF的产生在缺氧时增加了三倍，而糖尿病成纤维细胞VEGF的产生在缺氧条件下没有上调（P < 0.001）。为了解决这些差异是否由慢性高血糖症或缺乏瘦素受体引起的问题，在糖尿病发作之前（4至5周龄）从新生db/db小鼠收获成纤维细胞。这些成纤维细胞在基础或缺氧条件下VEGF的产生没有受损，证实了成熟小鼠中db/db成纤维细胞的结果是由糖尿病状态引起的，而不是因为瘦素-瘦素受体轴的改变。细胞活力的标志物，包括增殖和衰老，糖尿病和野生型成纤维细胞之间没有显着差异。我们的结论是，在体外，糖尿病成纤维细胞表现出选择性损伤离散的细胞过程中至关重要的组织修复，包括细胞迁移，血管内皮生长因子的生产，和缺氧的反应。VEGF异常与高血糖症的发作同时发生。并且在血糖正常的瘦素受体缺陷型db/db小鼠中没有观察到。这些观察结果支持成纤维细胞功能障碍在人类糖尿病患者中观察到的受损伤口愈合中的作用，并且还表明糖尿病患者缺血性损伤后发生的不良临床结局的机制。
Although it is known that systemic diseases such as diabetes result in impaired wound healing, the mechanism for this impairment is not understood. Because fibroblasts are essential for wound repair, we compared the in vitro behavior of fibroblasts cultured from diabetic, leptin receptor-deficient (db/db) mice with wild-type fibroblasts from mice of the same genetic background in processes important during tissue repair. Adult diabetic mouse fibroblast migration exhibited a 75% reduction in migration compared to normal fibroblasts (P < 0.001) and was not significandy stimulated by hypoxia (1% O-2), whereas wildtype fibroblast migration was up-regulated nearly twofold in hypoxic conditions (P < 0.05). Diabetic fibroblasts produced twice the amount of pro-matrix metalloproteinase-9 as normal fibroblasts, as measured by both gelatin zymography and enzyme-linked immunosorbent assay (P < 0.05). Adult diabetic fibroblasts exhibited a sevenfold impairment In vascular endothelial growth factor (VEGF) production (4.5 +/- 1.3 pg/ml versus 34.8 +/- 3.3 pg/ml, P < 0.001) compared to wild-type fibroblasts. Moreover, wild-type fibroblast production of VEGF increased threefold in response to hypoxia, whereas diabetic fibroblast production of VEGF was not up-regulated in hypoxic conditions (P < 0.001). To address the question whether these differences resulted from chronic hyperglycemia or absence of the leptin receptor, fibroblasts were harvested from newborn db/db mice before the onset of diabetes (4 to 5 weeks old). These fibroblasts showed no impairments in VEGF production under basal or hypoxic conditions, confirming that the results from db/db fibroblasts in mature mice resulted from the diabetic state and were not because of alterations in the leptin-leptin receptor axis. Markers of cellular viability including proliferation and senescence were not significantly different between diabetic and wild-type fibroblasts. We conclude that, in vitro, diabetic fibroblasts show selective impairments in discrete cellular processes critical for tissue repair including cellular migration, VEGF production, and the response to hypoxia. The VEGF abnormalities developed concurrently with the onset of hyperglycemia. and were not seen in normoglycemic, leptin receptor-deficient db/db mice. These observations support a role for fibroblast dysfunction in the impaired wound healing observed in human diabetics, and also suggest a mechanism for the poor clinical outcomes that occur after ischemic injury in diabetic patients.