Research on the mechanism of immune-mediated mitochondrial dysfunction in the pathogenesis of ITP-related fatigue and the regulating effect of Chinese formula JPYQSX
Description
Using murine C2C12 myoblasts as the cell model, the study was divided into two sections. In Experiment One, different concentrations of IL-6 or JPYQSX were administered to C2C12 myoblasts for 72 hours,and cell vitality changes were monitored at 24 hours, 48 hours,and 72 hours. The alterations in cell vitality were evaluated by CCK-8. Experiment Two was comprised of four groups: normal, IL-6, JPYQSX, and co-treated group. In Experiment Two,the expression of HIF-1α and SDH in C2C12 myoblasts was detected by Western blot. The content of ATP was detected by spectrophotometry. The levels of succinate and fumarate were detected by colorimetry. The content of ROS was detected by fluorescent probe technique.In Experiment One: At 24 hours, the cell vitality in each group treated with different concentrations of IL-6 was significantly higher than that in the control group. At 72 h, there was a statistically significant decrease in the cell vitality of the 20 ng/mL and 40 ng/mL IL-6 groups when compared to the control group. Additionally, myoblast vitality was negatively correlated with IL-6 intervention time and dose. In Experiment Two: Compared with the normal group,the protein expression of SDH and HIF-1α at 24 hours in the IL-6 group was significantly improved. The co-treated group had lower SDH and HIF-1α levels than the IL-6 group. Compared with the normal group,the level of ATP in the IL-6 group was significantly increased. Compared with the IL-6 group,the level of ATP in the co-treated group was lower. Compared with the normal group,the levels of succinate and fumarate in the IL-6 group were significantly increased. Compared with the IL-6 group,the levels of succinate and fumarate in the co-treated group were reduced significantly. Compared with the normal group,the level of ROS in the IL-6 group was significantly increased. After JPYQSX intervention,the level of ROS in the co-treated group was reduced significantly.