Human arterial smooth muscle cells rapidly deplete cell culture media of glucose.
Human arterial smooth muscle cells rapidly deplete cell culture media of glucose.
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人动脉平滑肌细胞迅速耗尽细胞培养基中的葡萄糖。
DOI：
10.1007/s001250100598
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发表时间：
2001
期刊：
影响因子：
8.2
通讯作者：
Bornfeldt,KE
中科院分区：
文献类型：
--
作者：
Renard,CB;Bornfeldt,KE
致编者：动脉平滑肌细胞有助于动脉粥样硬化和心血管疾病病变的进展。我们最近在一个新的猪糖尿病加速动脉粥样硬化模型中发现，动脉平滑肌细胞在动脉粥样硬化病变处的积累和增殖明显受到糖尿病的刺激。因此，增加对动脉平滑肌生物学的了解对糖尿病心血管并发症的研究具有重要意义。培养的动脉平滑肌细胞为这一领域的研究提供了一个有用的模型。葡萄糖的消耗速率在不同的细胞类型和组织之间差别很大。在这项研究中，我们使用未经转化的二倍体平滑肌细胞，通过外植体方法[1]从人主动脉分离，以确定这些细胞的葡萄糖消耗率。培养细胞传代过程中的葡萄糖消耗（3ą9）通过组织培养基中葡萄糖的损失来测量，使用标准酶法（Sigma, St. Louis, Mo.， USA）。只要葡萄糖浓度高于2 mmol/l，葡萄糖消耗速率在数天内呈线性（数据未显示）。结果表明，在含有10% FCS和5.6 mmol/l葡萄糖的DMEM中培养的增殖的人主动脉平滑肌细胞的葡萄糖消耗量高于增殖淋巴细胞[2]和许多其他类型的细胞。葡萄糖消耗量为；分别为0.54 0.02 pmol glucose´hą1´cellą1, n= 8)、0.62 0.06 n= 6)和0.63 0.03 pmol glucose´hą1´cellą1 n= 3)。N表示在一次有代表性的实验中分析每个供体的板数。动脉平滑肌的高葡萄糖消耗率与恶性细胞相似，在有氧条件下，恶性细胞的葡萄糖摄取和糖酵解率增加了约10倍[3,4]。有趣的是，动脉平滑肌在有氧条件下也具有较高的糖酵解率[1,5]。在这方面，培养的正常人动脉平滑肌细胞类似于肿瘤细胞和一些其他类型的细胞，如增殖淋巴细胞[2]和胸腺细胞[6]。具有高增殖能力的细胞所需增加的能量可能主要由糖酵解而不是氧化葡萄糖分解提供。在这种情况下，有人提出糖酵解虽然在ATP生产方面对细胞非常不利，但可以作为一种减少氧化应激的保护策略。人体动脉平滑肌细胞的高葡萄糖消耗率对研究不同葡萄糖浓度对这些细胞的影响具有重要意义。在相对较短的时间内，在24孔板中保存的细胞培养物在10% FCS（每孔1 ml培养基）的存在下迅速耗尽培养基中的葡萄糖，除非经常更换培养基。研究开始时含有5.6 mmol/l葡萄糖的培养基在培养1天后仅含有2.7 0.13 mmol/l葡萄糖，2天后完全无葡萄糖（图1）。人类动脉平滑肌细胞可以
To the Editor: Arterial smooth muscle cells contribute to the progression of lesions of atherosclerosis and cardiovascular disease. We have recently shown that accumulation and proliferation of arterial smooth muscle cells in atherosclerotic lesions are markedly stimulated by diabetes in a new porcine model of diabetes-accelerated atherosclerosis [1]. Thus, increased understanding of arterial smooth muscle biology is important for studies on cardiovascular complications of diabetes. Cultured arterial smooth muscle cells provide a useful model that is frequently used for studies in this area. The rate of glucose consumption varies widely between different cell types and tissues. In this study, we used non-transformed diploid smooth muscle cells isolated by an explant method [1] from the human aorta to determine glucose consumption rates in these cells. Glucose consumption in cultured cells passage 3ą9) was measured as loss of glucose from tissue culture media, using a standard enzymatic method Sigma, St. Louis, Mo., USA). The rate of glucose consumption was linear for several days as long as glucose concentrations were higher than 2 mmol/l data not shown). The results show that glucose consumption in proliferating human aortic smooth muscle cells incubated in DMEM containing 10% FCS and 5.6 mmol/l glucose is high compared to that of proliferating lymphocytes [2] and many other cell types. Glucose consumption is; 0.54 0.02 pmoles glucose ī hą1 ī cellą1 means SD, n= 8), 0.62 0.06 n= 6) and 0.63 0.03 pmoles glucose ī hą1 ī cellą1 n= 3), respectively, in human smooth muscle cells isolated from three donors. n represents the number of plates analysed from each donor in one representative experiment. The high rate of glucose consumption in arterial smooth muscle is similar to that of malignant cells that are known to exhibit about a tenfold increase in glucose uptake and high glycolysis under aerobic conditions [3, 4]. Of interest, arterial smooth muscle also has a high rate of glycolysis under aerobic conditions [1, 5]. In this respect normal human arterial smooth muscle cells in culture resemble tumour cells and a few other cell types, eg proliferating lymphocytes [2] and thymocytes [6]. It is possible that the increased energy required in cells with a high proliferative capacity is supplied primarily by glycolysis rather than by oxidative glucose breakdown. In this context, it has been suggested that glycolysis, although highly unfavourable for the cell in terms of ATP production, could serve as a protective strategy to minimize oxidative stress [6]. The high rate of glucose consumption in human arterial smooth muscle cells has implications for studies of effects of different glucose concentrations in these cells. During a relatively short period of time, cell cultures kept in 24-well plates in the presence of 10% FCS 1 ml medium per well) rapidly deplete the medium of glucose unless the medium is replaced frequently. A medium that contains 5.6 mmol/l glucose at the beginning of the study contains only 2.7 0.13 mmol/l glucose after 1 day of incubation and is completely devoid of glucose after 2 days Fig. 1). Human arterial smooth muscle cells do