Ribeiro Tiago, Reis Mariana, Vasconcelos Vitor, Urbatzka Ralph
Interdisciplinary Centre of Marine and Environmental Research (CIIMAR/CIMAR), University of Porto, Avenida General Norton de Matos, s/n, Matosinhos, 4450-208, Portugal.
Faculty of Sciences, University of Porto, Rua do Campo Alegre, Porto, 1021, 4169-007, Portugal.
Sci Rep. 2024 Dec 30;14(1):32142. doi: 10.1038/s41598-024-83986-0.
Diabetes is a pandemic disease that causes the loss of control of glucose regulation in the organism, in consequence of dysfunction of insulin production or functionality. In this work, the antidiabetic bioactivity of 182 fractions from 19 cyanobacteria strains derived from the LEGE Culture Collection were analysed using the 2-NBDG assay in zebrafish larvae. From this initial screening, two fractions (57 (06104_D) and 107 (03283_B)) were identified as promising insulin mimetics. These were further characterized by measuring glucose levels in whole larvae, the expression of glucose transporters (GLUT 1-3) using western blot, and the mRNA expression levels of the glut2, pepck, and insa genes using real-time qPCR. Both fractions showed a decrease in free glucose levels. Furthermore, exposure to fraction 06104_D decreased GLUT1 and increased insa mRNA levels. The chemical composition of these fractions was determined using LC-HRESIMS/MS and compared to inactive fractions of the same polarity in order to identify the unique bioactive molecules. The molecular networks constructed using the GNPS platform revealed that fraction 06104_D contained mass clusters primarily composed of chlorins, lipids, and terpenoids, while fraction 03283_B contained xanthophylls, peptides, and terpenoids. To correlate the observed activity with the chemical composition of fraction 06104_D, pheophorbide a was chosen as a representative of chlorophyll derivatives. Exposure to zebrafish larvae at 10 and 20 µM confirmed the increased glucose uptake on the 2-NBDG assay. These findings highlight the bioactivity of chlorophyll derivatives as insulin mimetic compounds, as well as cyanobacteria as a source of potential therapeutic diabetes applications.
糖尿病是一种大流行性疾病，由于胰岛素产生或功能障碍，导致机体葡萄糖调节失控。在这项研究中，使用2-NBDG测定法对来自LEGE培养物保藏中心的19株蓝藻菌株的182个组分的抗糖尿病生物活性进行了分析。从最初的筛选中，确定了两个组分（57（06104_D）和107（03283_B））为有前景的胰岛素模拟物。通过测量整个幼虫的葡萄糖水平、使用蛋白质印迹法检测葡萄糖转运蛋白（GLUT 1-3）的表达以及使用实时定量PCR检测glut2、pepck和insa基因的mRNA表达水平，对这些组分进行了进一步表征。两个组分均显示游离葡萄糖水平降低。此外，暴露于组分06104_D会降低GLUT1并增加insa mRNA水平。使用LC-HRESIMS/MS测定了这些组分的化学成分，并与相同极性的无活性组分进行比较，以鉴定独特的生物活性分子。使用GNPS平台构建的分子网络显示，组分06104_D包含主要由二氢卟吩、脂质和萜类化合物组成的质量簇，而组分03283_B包含叶黄素、肽和萜类化合物。为了将观察到的活性与组分06104_D的化学成分相关联，选择脱镁叶绿酸a作为叶绿素衍生物的代表。在10和20 µM浓度下暴露于斑马鱼幼虫，证实了在2-NBDG测定中葡萄糖摄取增加。这些发现突出了叶绿素衍生物作为胰岛素模拟化合物的生物活性，以及蓝藻作为潜在治疗糖尿病应用来源的作用。
