Department of Human Morphology Città Studi, Università degli Studi di Milano, Milan, Italy.
Nutr J. 2011 Jul 25;10:77. doi: 10.1186/1475-2891-10-77.
Micronutrient inadequate intake is responsible of pathological deficiencies and there is a need of assessing the effectiveness of metal supplementation, frequently proposed to rebalance poor diets. Manganese (Mn) is present in many enzymatic intracellular systems crucial for the regulation of cell metabolism, and is contained in commercially available metal supplements.
We compared the effects of two different commercial Mn forms, gluconate (MnGluc) and oxyprolinate (MnOxP). For this purpose we used the polarized Caco-2 cells cultured on transwell filters, an established in vitro model of intestinal epithelium. Since micronutrient deficiency may accelerate mitochondrial efficiency, the mitochondrial response of these cells, in the presence of MnGluc and MnOxP, by microscopy methods and by ATP luminescence assay was used.
In the presence of both MnOxP and MnGluc a sustained mitochondrial activity was shown by mitoTraker labeling (indicative of mitochondrial respiration), but ATP intracellular content remained comparable to untreated cells only in the presence of MnOxP. In addition MnOxP transiently up-regulated the antioxidant enzyme Mn superoxide dismutase more efficiently than MnGluc. Both metal treatments preserved NADH and βNADPH diaphorase oxidative activity, avoided mitochondrial dysfunction, as assessed by the absence of a sustained phosphoERK activation, and were able to maintain cell viability.
Collectively, our data indicate that MnOxP and MnGluc, and primarily the former, produce a moderate and safe modification of Caco-2 cell metabolism, by activating positive enzymatic mechanisms, thus could contribute to long-term maintenance of cell homeostasis.
微量营养素摄入不足会导致病理性缺乏,因此需要评估金属补充的有效性,金属补充通常被提议用于重新平衡不良饮食。锰(Mn)存在于许多细胞内酶系统中,这些酶系统对细胞代谢的调节至关重要,并且存在于市售的金属补充剂中。
我们比较了两种不同的商业 Mn 形式,葡萄糖酸盐(MnGluc)和羟脯氨酸盐(MnOxP)的作用。为此,我们使用在 Transwell 过滤器上培养的极化 Caco-2 细胞,这是一种成熟的肠上皮体外模型。由于微量营养素缺乏可能会加速线粒体效率,因此我们使用显微镜方法和 ATP 发光测定法研究了这些细胞在 MnGluc 和 MnOxP 存在下的线粒体反应。
在 MnOxP 和 MnGluc 的存在下,mitoTraker 标记(表明线粒体呼吸)显示出持续的线粒体活性,但只有在 MnOxP 的存在下,细胞内的 ATP 含量才保持与未处理细胞相当。此外,MnOxP 比 MnGluc 更有效地瞬时上调抗氧化酶 Mn 超氧化物歧化酶。两种金属处理均能保持 NADH 和 βNADPH 二氢还原酶的氧化活性,避免线粒体功能障碍,如持续的磷酸化 ERK 激活缺失所评估的那样,并能够维持细胞活力。
总的来说,我们的数据表明,MnOxP 和 MnGluc,尤其是前者,通过激活积极的酶促机制,对 Caco-2 细胞代谢产生适度和安全的修饰,从而有助于长期维持细胞内稳态。