Evaluation of Reactive Oxygen Species Production in Human Adipose Tissue-Derived Mesenchymal Stem Cells under High D-Glucose Condition

Type 2 diabetes mellitus (T2DM) is viewed as a serious medical condition due to its microvascular and macrovascular complications, which are characterized by insulin resistance (IR). Reactive oxygen species (ROS) are considered to be the factor that provokes insulin resistance in T2DM. To examine the pathological mechanism of T2DM, AT-MSCs have been used to establish a disease model mimicking insulin resistance using high D-glucose. SM@siRNP has also been employed to investigate the ROS reduction effect on D-glucose-treated AT-MSCs. The expression of insulin resistance-related genes and EGR-1 protein were analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The ROS release was identified by DCFH-DA assay. The results demonstrated that high D-glucose (100 mM) has significantly increased the expression of insulin resistance-related genes (EGR-1, PTEN, GGPS-1) (p < 0.01), as well as the production of intracellular ROS (p > 0.001), compared to the non-treated group. SM@siRNP has been shown to decrease ROS production by antioxidant capacity and anti-inflammatory efficacy (p < 0.01). However, to confirm the effect of SM@siRNP on improving insulin sensitivity in AT-MSCs, the expression of those insulin resistance-related genes after medication treatment must be identified. In this study, the results provided an aspect of insulin resistance pathological mechanism which can be applied for further research on T2DM treatment, as well as an in vitro model for pre-clinical drug screening that can be used instead of an in vivo model.