Moskot Marta, Jakóbkiewicz-Banecka Joanna, Kloska Anna, Smolińska Elwira, Mozolewski Paweł, Malinowska Marcelina, Rychłowski Michał, Banecki Bogdan, Węgrzyn Grzegorz, Gabig-Cimińska Magdalena
Laboratory of Molecular Biology (affiliated with the University of Gdańsk), Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Wita Stwosza 59, 80-308 Gdańsk, Poland.
Department of Molecular Biology, University of Gdańsk, Wita Stwosza 59, 80-308 Gdańsk, Poland.
Sci Rep. 2015 Mar 23;5:9378. doi: 10.1038/srep09378.
Flavonoids were found previously to modulate efficiency of synthesis of glycosaminoglycans (GAGs), compounds which are accumulated in cells of patients suffering from mucopolysaccharidoses (MPSs). The aim of this work was to determine effects of different flavonoids (genistein, kaempferol, daidzein) used alone or in combinations, on expression of genes coding for proteins involved in GAG metabolism. Analyses with DNA microarray, followed by real-time qRT-PCR revealed that genistein, kaempferol and combination of these two compounds induced dose- and time-dependent remarkable alterations in transcript profiles of GAG metabolism genes in cultures of wild-type human dermal fibroblasts (HDFa). Interestingly, effects of the mixture of genistein and kaempferol were stronger than those revealed by any of these compounds used alone. Similarly, the most effective reduction in levels of GAG production, in both HDFa and MPS II cells, was observed in the presence of genistein, keampferol and combination of these compounds. Forty five genes were chosen for further verification not only in HDFa, but also in MPS II fibroblasts by using real-time qRT-PCR. Despite effects on GAG metabolism-related genes, we found that genistein, kaempferol and mixture of these compounds significantly stimulated expression of TFEB. Additionally, a decrease in MTOR transcript level was observed at these conditions.
先前发现黄酮类化合物可调节糖胺聚糖(GAGs)的合成效率,糖胺聚糖是黏多糖贮积症(MPSs)患者细胞中积累的化合物。这项工作的目的是确定单独使用或联合使用不同黄酮类化合物(染料木黄酮、山奈酚、大豆苷元)对参与GAG代谢的蛋白质编码基因表达的影响。通过DNA微阵列分析,随后进行实时定量逆转录聚合酶链反应(qRT-PCR),结果显示染料木黄酮、山奈酚以及这两种化合物的组合在野生型人皮肤成纤维细胞(HDFa)培养物中诱导了GAG代谢基因转录谱的剂量和时间依赖性显著变化。有趣的是,染料木黄酮和山奈酚混合物的作用比单独使用这两种化合物中的任何一种都更强。同样,在染料木黄酮、山奈酚以及它们的组合存在的情况下,在HDFa和MPS II细胞中均观察到GAG产生水平的最有效降低。通过实时qRT-PCR选择了45个基因,不仅在HDFa中,而且在MPS II成纤维细胞中进行进一步验证。尽管对GAG代谢相关基因有影响,但我们发现染料木黄酮、山奈酚以及它们的混合物显著刺激了转录因子EB(TFEB)的表达。此外,在这些条件下观察到雷帕霉素靶蛋白(MTOR)转录水平降低。