Bioassay Guided Partition of Dandang Gendis (Clinacanthus nutans L.) Monitored by Glucose Consumption Assay on Myoblast Cells and GLUT4 Molecular Docking Analysis

Abstract

Diabetes mellitus (DM) is a prevalent global metabolic disorder, with an increasing incidence in Indonesia. Clinacanthus nutans L. has been traditionally used for its hypoglycemic properties, yet its precise mechanism of action remains unclear. This study aimed to assess the effects of C. nutans partitions on glucose consumption and to predict their potential interaction with GLUT4 through in silico analysis. A bioassay-guided partition approach was employed to partition C. nutans. Insulin resistance was induced in C2C12 myoblasts using 0.75 mM palmitic acid, followed by treatment with C. nutans partitions at concentrations of 500, 250, and 125 μg/mL. Glucose consumption was quantified using the GOD/PAP assay, with statistical significance at p<0.05. Moreover, molecular docking analysis was conducted to evaluate the binding affinity of C. nutans bioactive compounds to GLUT4. Results indicated that the insoluble partition of C. nutans significantly enhanced glucose uptake in insulin-resistant C2C12 cells at 500 µg/mL (p = 0.0021) and 250 µg/mL (p = 0.0198) compared to untreated controls. However, molecular docking analysis did not reveal direct binding interactions between C. nutans compounds and GLUT4. In conclusion, the insoluble partition of C. nutans effectively improves glucose consumption in insulin-resistant C2C12 cells, though its mechanism of action does not appear to involve direct GLUT4 modulation. These findings suggest that C. nutans holds promise as a complementary therapy for insulin resistance, aligning with Sustainable Development Goal (SDG) 3 by contributing to alternative treatments for type 2 diabetes mellitus (T2DM) and SDG 9 by fostering pharmaceutical innovation and sustainable plant-based therapies.