Antidiabetic activities of Bolanthus spergulifolius (Caryophyllaceae) extracts on insulin-resistant 3T3-L1 adipocytes

• Published in: PloS one 2021-06, Vol.16 (6), p.e0252707-e0252707
• Main Authors: Derici, Gizem Ece, Özdas, Sibel, Canatar, Ipek, Koç, Murat
• Format: Article
• Language: English
• Subjects: Adipocytes; Adipose tissue; Antibiotics; Antidiabetics; Apoptosis; Bioengineering; Biology and Life Sciences; Blood glucose; Calibration; Care and treatment; Complications; Computer and Information Sciences; Diabetes; Diabetes mellitus; Diabetes therapy; Endocrine secretions; Engineering and Technology; Enzymes; Fat cells; Gallic acid; Glucosidase; Health aspects; Hyperglycemia; Hypoglycemic agents; Insulin; Insulin resistance; Integrative medicine; Materia medica, Vegetable; Medicine and Health Sciences; Metabolism; Patient outcomes; Pharmacology, Experimental; Phenolic compounds; Phenols; Physical Sciences; Plant extracts; Public health; Reagents; Research and Analysis Methods; Secretions; α-Glucosidase
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0252707

Diabetes mellitus (DM) is a metabolic disorder with chronic hyperglycemia featured by metabolic outcomes owing to insufficient insulin secretion and/or insulin effect defect. It is critical to investigate new therapeutic approaches for T2DM and alternative, natural agents that target molecules in potential signal pathways. Medicinal plants are significant resources in the research of alternative new drug active ingredients. Bolanthus spergulifolius (B . spergulifolius) is one of the genera of the family Caryophyllaceae. In this study, it was explored the potential anti-diabetic effects in vitro of B . spergulifolius extracts on 3T3-L1 adipocytes. The total phenolic contents (TPC) of methanolic (MeOH), ethyl acettate (EA) and aqueous extracts of B . spergulifolius were evaluated via Folin-Ciocateau. B . spergulifolius extracts showing highly TPC (Aqueous< MeOH< EA) and their different concentrations were carried out on preadipocytes differentiated in to mature 3T3-L1 adipocytes to investigate their half-maximal (50%) inhibitory concentration (IC 50 ) value by using Thiazolyl blue tetrazolium bromide (MTT) assay. The IC 50 of MeOH, EA and Aqueous extracts were observed as 305.7 ± 5.583 μg/mL, 567.4 ± 3.008 μg/mL, and 418.3 ± 4.390 μg/mL and used for further experiments. A live/dead assay further confirmed the cytotoxic effects of MeOH, EA and Aqueous extracts (respectively, 69.75 ± 1.70%, 61.75 ± 1.70%, 70 ± 4.24%, and for all p< 0.05). Also, effects of extracts on lipid accumulation in mature 3T3-L1 adipocytes were evaluated by Oil-Red O staining assay. The extracts effectively decreased lipid-accumulation compared to untreated adipocytes (for all p< 0.05). Moreover, effect of extracts on apoptosis regulated by the Bax and Bcl-2 was investigated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The extracts significantly induced apoptosis by up-regulating pro-apoptotic Bax expression but down-regulated anti-apoptotic Bcl-2 gene expression compared to untreated adipocytes (for all p< 0.05). The Glut-4 expression linked with insulin resistance was determined by qRT-PCR, Western-blot analysis, and immunofluorescence staining. In parallel, the expression of Glut-4 in adipocytes treated with extracts was significantly higher compared to untreated adipocytes (for all p< 0.05). Extracts significantly suppressed cell migration after 30 h of wounding in a scratch-assay (for all p< 0.05). Cell morphology and diameter were further evaluated by