Investigating the Preventive Effects of Tinospora crispa and Tinospora cordifolia on Methylglyoxal-induced Cytotoxicity and Metabolic Dysregulation in Osteoblast Cells

LUQMAN, JAYA (0202) Investigating the Preventive Effects of Tinospora crispa and Tinospora cordifolia on Methylglyoxal-induced Cytotoxicity and Metabolic Dysregulation in Osteoblast Cells. Masters thesis, UNIMAS.

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Abstract

Patients with type 2 mellitus (T2DM) typically exhibit significantly increased methylglyoxal (MGO) levels. MGO is a reactive carbonyl compound and a byproduct of glycolysis, serving as a precursor of advanced glycation end products (AGE). AGE enters cells via its receptor of advanced glycation end products (RAGE) leading to oxidative stress, inflammation and consequently death of the cells. In osteoblasts, high AGE level can induce inflammatory response and oxidative stress (OS), as well as reducing osteoblastogenesis. Tinospora cordifolia (T. cordifolia) and Tinospora crispa (T. crispa) have been traditionally used to treat diabetes. In modern medicine, metformin serves as the primary treatment to reduce hyperglycaemia when lifestyle changes have failed. Currently, little is known about the effects T. cordifolia and T. crispa on bone remodelling when bone cells are exposed to high MGO levels. In addition, there is no data available on the interaction of metformin with T. crispa and T. cordifolia extracts. Therefore, this study aimed to evaluate the effects of T. crispa and T. cordifolia on bone metabolism in MGO-stimulated osteoblast cells (hFOB1.19) and explore the potential synergistic interactions of these two plants with metformin. The plants underwent extraction using methanol and hot water (aqueous). The phytochemical content of the four extracts were determined using GC-MS, the antioxidant activities were assessed via DPPH assay while anti-glycation properties were assessed via BSA-MGO assay. Then, the effects of the extracts on MGO-stimulated osteoblast cells were assessed with or without metformin for antioxidant capacity, anti-apoptotic activity, and anti-inflammatory properties as well as the impact on the expression and activity of the osteoblastic insulin receptors. The results showed that the plant extracts contained antioxidant and anti-inflammatory bioactive compounds, with notable scavenging activity and anti-apoptotic effects. However, the extracts showed limited anti-glycation activity as shown in BSA-MGO and AGE formation in MGO-stimulated osteoblasts. Furthermore, the extracts significantly reduced the secretion of IL-6 and IL-18 suggesting their anti-inflammatory properties. The extracts also increased the expression of Runx2 in influencing the osteoblastogenesis. In addition, they increased expression of GLUT4, and insulin receptor in MGO-stimulated hFOB1.19, suggesting positive effects on diabetic bone metabolism. In conclusion, this study highlights the potential of T. cordifolia and T. crispa extracts in mitigating T2DM-related complications, particularly bone loss.

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