Sains Malaysiana 52(10)(2023): 2815-2827

http://doi.org/10.17576/jsm-2023-5210-07

Purified Polysaccharides Extracted from Grey Oyster Mushroom [Pleurotus sajor-caju (Fr.) Sing.]  Stimulate Glucose Uptake in C2C12 Myotubes through the activation of AMP-Activated Protein Kinase (AMPK) and Glucose Transporter 1 (GLUT1) Proteins

(Polisakarida Dimurnikan Diekstrak daripada Cendawan Tiram Kelabu [Pleurotus sajor-caju (Fr.) Sing.] Merangsang Pengambilan Glukosa dalam Miotiub C2C12 melalui Pengaktifan Protein Kinase Diaktifkan AMP (AMPK) dan Protein Pengangkut Glukosa 1 (GLUT1))

KULWANIT PATNINAN¹, DECHA SERMWITTAYAWONG^1,*, SUPADA NUINAMWONG¹, WIPAPAN KHIMMAKTONG¹, KUSUMARN NOIPHA² & NONGPORN HUTADILOK-TOWATANA¹

¹Division of Health and Applied Sciences, Faculty of Science, Prince of Songkla University, Hatyai, Songkhla 90110, Thailand

²Faculty of Health and Sports Science, Thaksin University, Paphayom, Phatthalung, 93110

Thailand

Received: 10 May 2023/Accepted: 7 September 2023

Abstract

The grey oyster mushroom [Pleurotus sajor-caju (Fr.) Sing.], an edible mushroom, has been known as a source of bioactive compounds, including polysaccharides.  Polysaccharides from this mushroom have been shown to possess antidiabetic activity both in vivo and in vitro.  However, antidiabetic mechanism of partially purified or purified polysaccharides from the gray oyster mushroom has not been characterized. In this study, we extracted and purified polysaccharides from gray oyster mushrooms and used them to investigate the antidiabetic mechanism in the context of C2C12 myotubes.  Using Fourier Transform Infrared spectroscopy (FTIR) analysis and enzymatic assay, we showed that the polysaccharide sample, namely 9S1-1, contains b-glucose, a-glucose, and mannose as the monosaccharide composition, and b-glucan is the major type of polysaccharide in the sample. This 9S1-1 sample dose-dependently stimulated glucose uptake in C2C12 myotubes. Further analysis showed that the sample activated the AMP-activated protein kinase (AMPK) but not Akt serine/threonine kinase (AKT) phosphorylation, suggesting that the stimulation is AMPK-dependent. Moreover, we showed that compound c, an inhibitor of AMPK, inhibited glucose uptake in 9S1-1-stimulated cells, confirming the requirement of AMPK in the glucose uptake activated by the 9S1-1 sample.  In addition, it promoted glucose transporter protein type 1 (GLUT1) but not GLUT4 protein expression. These results suggest that GLUT1 may be responsible for the stimulation of glucose uptake in 9S1-1-activated cells. Together, these data illustrate the antidiabetic mechanism of polysaccharides isolated from the gray oyster mushroom and the potential use of the polysaccharide as an antidiabetic agent. 

Keywords: AMPK; antidiabetic; GLUT1; Gray oyster mushroom; Pleurotus sajor-caju

Abstract

Cendawan tiram kelabu [Pleurotus sajor-caju (Fr.) Sing.], cendawan yang boleh dimakan, telah dikenali sebagai sumber sebatian bioaktif, termasuk polisakarida. Polisakarida daripada cendawan ini telah terbukti mempunyai aktiviti antidiabetis secara in vivo dan in vitro. Walau bagaimanapun, mekanisme antidiabetis bagi polisakarida yang telah dimurnikan sebahagian atau dimurnikan daripada cendawan tiram kelabu belum dicirikan. Dalam kajian ini, kami mengekstrak dan memurnikan polisakarida daripada cendawan tiram kelabu dan menggunakannya untuk mengkaji mekanisme antidiabetis dalam konteks miotiub C2C12. Dengan menggunakan analisis spektroskopi Transformasi Fourier Inframerah (FTIR) dan asai enzim, kami menunjukkan bahawa sampel polisakarida, iaitu 9S1-1, mengandungi b-glukosa, a-glukosa dan manosa sebagai komposisi monosakarida dan b-glukan ialah jenis utama polisakarida dalam sampel. Sampel 9S1-1 ini merangsang pengambilan glukosa dos berkeperluan dalam miotiub C2C12. Analisis lanjut menunjukkan bahawa sampel mengaktifkan protein kinase diaktifkan AMP (AMPK) tetapi bukan fosforilasi Akt serin/threonina kinase (AKT), mencadangkan bahawa rangsangan adalah sandaran AMPK. Selain itu, kami menunjukkan bahawa kompaun c, iaitu perencat AMPK, merencat pengambilan glukosa dalam sel yang dirangsang 9S1-1, mengesahkan keperluan AMPK dalam pengambilan glukosa yang diaktifkan oleh sampel 9S1-1. Di samping itu, ia menggalakkan protein pengangkut glukosa jenis 1 (GLUT1) tetapi bukan ekspresi protein GLUT4. Keputusan ini menunjukkan bahawa GLUT1 mungkin bertanggungjawab untuk rangsangan pengambilan glukosa dalam sel yang diaktifkan 9S1-1. Data ini menggambarkan mekanisme antidiabetis polisakarida yang dipencilkan daripada cendawan tiram kelabu dan potensi penggunaan polisakarida sebagai agen antidiabetis.

Kata kunci: AMPK; antidiabetis; Cendawan tiram kelabu; GLUT1; Pleurotus sajor-caju

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*Corresponding author; email: decha.s@psu.ac.th