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Sains Malaysiana 54(12)(2025): 2907-2916
http://doi.org/10.17576/jsm-2025-5412-09
Quantitative
ELISA Analysis of Protein Expression in HepG2 Cells Following Treatment with
Insulin, Glucose, and Bis(maltolato)oxovanadium(IV)
(Analisis Kuantitatif ELISA Pengekspresan Protein dalam Sel HepG2 Selepas Rawatan dengan Insulin, Glukosa dan Bis(maltolato)oksovanadium(IV))
Received: 19 December 2024/Accepted: 8 September 2025
Abstract
Type
2 diabetes, the most common form of diabetes mellitus, is caused by insulin
resistance. Many studies have sought orally active therapeutic compounds to
replace insulin injections. This study examines the expression levels of key
proteins involved in the insulin receptor (InsR) and
AMP-activated protein kinase (AMPK) signaling pathways in HepG2 cells using
ELISA, following treatment with insulin, glucose, and/or bis(maltolato)oxovanadium(IV) (BMOV). Treatment
with BMOV for 24 h in the presence of glucose significantly increased the
levels of glucose transporter type 4 (GLUT4) in the cell lysate from 511 ± 7
ng/L (control) to 882 ± 5 ng/L, and AMPK α1 in the supernatant from 683 ±
9 ng/L to 789 ± 6 ng/L. BMOV treatment for 1 h with glucose increased InsR levels in the lysate from 1.52 ± 0.11 ng/L to 4.01 ±
0.03 ng/L. Additionally, cell-based ELISA showed that AMPK β1 absorbance
increased significantly under glucose + BMOV (1 h) treatment (1.32 ± 0.04)
compared to the control (0.83 ± 0.03). These findings suggest that BMOV
modulates glucose-related signaling pathways and may target the AMPK system - a
major therapeutic axis for obesity, diabetes, and metabolic syndrome.
Keywords: AMP-activated
protein kinase; BMOV; diabetes; ELISA; insulin receptor
Abstrak
Diabetes jenis 2 adalah bentuk diabetes
mellitus yang paling biasa yang disebabkan oleh rintangan insulin. Banyak kajian telah mencari sebatian terapeutik aktif secara oral untuk menggantikan suntikan insulin. Penyelidikan ini mengkaji tahap pengekspresan protein utama yang terlibat dalam reseptor insulin (InsR) dan laluan isyarat protein kinase
(AMPK) diaktifkan AMP dalam sel HepG2 menggunakan ELISA, selepas rawatan dengan insulin, glukosa dan/atau bis(maltolato)oksovanadium(IV)
(BMOV). Rawatan dengan BMOV selama 24 jam dengan kehadiran glukosa dengan ketara meningkatkan tahap pengangkut glukosa jenis 4 (GLUT4) dalam sel lisat daripada 511 ± 7 ng/L (kawalan) kepada 882 ± 5 ng/L dan AMPK α1 dalam supernatan daripada 683 ± 9 ng/L kepada 789 ±
6 ng/L. Rawatan BMOV selama 1 jam dengan glukosa meningkatkan tahap InsR dalam lisat daripada 1.52 ± 0.11 ng/L kepada 4.01 ± 0.03 ng/L. Tambahan pula, ELISA berasaskan sel menunjukkan bahawa penyerapan AMPK β1 meningkat dengan ketara di bawah rawatan glukosa + BMOV (1 jam) (1.32 ± 0.04) berbanding kawalan (0.83 ± 0.03). Penemuan ini mencadangkan bahawa BMOV memodulasi laluan isyarat berkaitan glukosa dan mungkin menyasarkan sistem AMPK-paksi terapeutik utama untuk obesiti, diabetes dan sindrom metabolik.
Kata kunci: BMOV; diabetes; ELISA; kinase protein diaktifkan AMP; reseptor insulin
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*Corresponding author; email: a.alqatati@ju.edu.jo
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