الفهرس | Only 14 pages are availabe for public view |
Abstract In this current study, we evaluated the influence of BMMSC-CM on the HepG2 cell line and attempted to describe the possible underlying molecular signaling pathways involved. We used male albino rats for the isolation of BMMSCs. Then we identified those cells using culture morphology and specific surface markers of BMMSCs (CD44, CD90, and CD105) by flowcytometry. We then prepared the BMMSCs-CM to be used in this study. We treated HepG2 cells with multiple concentrations (20%, 40%, 60%, 80%, and 100%) of BMMSCs-CM over different durations of time (24 hours, 48hrs, and 72hrs). The MTT assay was used to determine the viability of HepG2 cells after BMMSCs-CM treatment, and it showed significantly decreased viability as the concentration was increased and as the time of treatment was prolonged. Flowcytometric analysis of apoptosis revealed an increased apoptotic rate 72 with cell cycle halt in the G0/G1 phase along with an inhibitory entry in the S phase. RT-PCR evaluation of P53 and Bcl-2 mRNA levels showed the upregulation of P53 and the downregulation of Bcl-2. Furthermore, protein level analysis of P53 and Bcl-2 by flowcytometry and western blotting revealed increased P53 and decreased Bcl-2 protein levels. In addition, ELISA results revealed increased ROS production and decreased TLR4 expression. |