Inhibited proliferation of fibroblasts derived from chronic diabetic wounds and normal dermal fibroblasts treated with high glucose is associated with increased formation L-lactate
Inhibited proliferation of fibroblasts derived from chronic diabetic wounds and normal dermal fibroblasts treated with high glucose is associated with increased formation L-lactate
复制标题
DOI：
10.1046/j.1524-475x.1998.60207.x
复制
发表时间：
1998-03-01
影响因子：
2.9
通讯作者：
Hansson, Anders
中科院分区：
文献类型：
--
作者：
Hehenberger, Karin;Heilborn, Johan D.;Hansson, Anders
糖尿病伴有伤口愈合延迟和肉芽组织形成不足，可能是由于成纤维细胞功能的缺陷。我们之前的研究表明，来自慢性糖尿病足溃疡的成纤维细胞比来自未受伤皮肤的成纤维细胞增殖更低。本研究旨在探讨非胰岛素依赖型糖尿病慢性伤口成纤维细胞和高糖培养的正常成纤维细胞增殖受损的可能机制。研究了两组患者的成纤维细胞：非糖尿病合并慢性静脉淤积性溃疡患者和非胰岛素依赖型糖尿病合并慢性糖尿病伤口患者。来自同一患者的未损伤皮肤和伤口活检作为成纤维细胞的来源。采用荧光法测定DNA含量，采用乳酸氧化酶分光光度法测定乳酸水平。我们发现加入非胰岛素依赖型糖尿病伤口成纤维细胞的条件培养基对正常成纤维细胞增殖有剂量依赖性的抑制作用。从这些细胞中提取的条件培养基显示L-乳酸水平升高，为6.3 +- 0.7 mmol/L，而非糖尿病细胞提取的培养基为2.1 +- 0.3 mmol/L (p < 0.01)，糖尿病未损伤皮肤成纤维细胞提取的培养基为3.5 +- 0.6 mmol/L，慢性非糖尿病伤口成纤维细胞提取的培养基为2.9 +- 0.3 mmol/L。在未损伤的正常成纤维细胞中添加6 mmol/L L-乳酸可降低DNA含量（58 +- 7%,p < 0.01）。以前我们已经表明，高葡萄糖浓度抑制成纤维细胞增殖并诱导生长因子抵抗。当培养基中d -葡萄糖的量增加时，所有细胞类型的l -乳酸水平都增加了。未损伤的正常细胞经β -羟基丁酸处理后，总DNA含量降低42 +- 5% (p < 0.05)， l -乳酸水平无显著升高。这些观察结果表明，l -乳酸的产生可能对体外成纤维细胞增殖很重要，并可能在体内成纤维细胞增殖中发挥作用。
Diabetes is accompanied by delayed wound healing and insufficient granulation tissue formation, possibly because of a defect in fibroblast function. We have previously shown that fibroblasts derived from chronic diabetic foot ulcers have lower proliferation compared with those from uninjured skin. The aim of this study was to investigate possible mechanisms explaining the impaired fibroblast proliferation observed in fibroblasts from non-insulin-dependent diabetes mellitus chronic wounds and normal fibroblasts cultured in high glucose. Fibroblasts from two groups of patients were studied: nondiabetic patients with chronic venous stasis ulcers and non-insulin-dependent diabetes mellitus patients with chronic diabetic wounds. Biopsies from both uninjured skin and wounds were taken from the same patients to serve as sources of fibroblasts. A fluorometric method was used to determine DNA content, and a spectrophotometric lactate oxidase method was used for lactate level analysis. We found a dose-dependent inhibition of normal fibroblast proliferation when adding conditioned media from non-insulin-dependent diabetes mellitus wound fibroblasts. The conditioned medium, from these cells showed elevated L-lactate levels, 6.3 +- 0.7 mmol/L compared with media derived from nondiabetic, 2.1 +- 0.3 mmol/L (p < 0.01), and diabetic uninjured skin fibroblasts, 3.5 +- 0.6 mmol/L and from chronic nondiabetic wound fibroblasts 2.9 +- 0.3 mmol/L. Addition of 6 mmol/L L-lactate to uninjured normal fibroblasts resulted in decreased DNA content (58 +- 7%, p < 0.01). Previously we have shown that high glucose concentrations inhibit fibroblast proliferation and induce growth factor resistance. When increasing the amount of D-glucose in the media, L-lactate levels increased in all cell types. When the uninjured normal cells were treated with beta-hydroxybutyrate, the total DNA content decreased by 42 +- 5% (p < 0.05), with no significant increase in the L-lactate levels. These observations indicate that L-lactate production may be of importance for fibroblast proliferation in vitro and may play a role in fibroblast proliferation in vivo.