Research on the mechanism of immune-mediated mitochondrial dysfunction in the pathogenesis of ITP-related fatigue and the regulating effect of Chinese formula JPYQSX

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.

以小鼠C2C12肌原细胞作为细胞模型，本研究分为两个部分。在实验一，不同浓度的IL-6或JPYQSX被施用于C2C12肌原细胞，持续72小时，并在24小时、48小时和72小时分别监测细胞活力变化。细胞活力的变化通过CCK-8进行评估。实验二分为四组：正常组、IL-6组、JPYQSX组和联合处理组。在实验二中，通过Western blot技术检测C2C12肌原细胞中HIF-1α和SDH的表达。ATP的含量通过分光光度法进行检测。琥珀酸和延胡索酸的水平通过比色法进行检测。活性氧（ROS）的含量通过荧光探针技术进行检测。在实验一： 在24小时时，与对照相比，接受不同浓度IL-6处理的各组细胞活力显著提高。 在72小时时，与对照相比，20 ng/mL和40 ng/mL IL-6组的细胞活力出现统计学上的显著下降。此外，肌原细胞活力与IL-6干预时间和剂量呈负相关。在实验二： 与正常组相比，IL-6组在24小时的SDH和HIF-1α蛋白表达显著增强。联合处理组的SDH和HIF-1α水平低于IL-6组。 与正常组相比，IL-6组的ATP水平显著升高。与IL-6组相比，联合处理组的ATP水平较低。 与正常组相比，IL-6组的琥珀酸和延胡索酸水平显著增加。与IL-6组相比，联合处理组的琥珀酸和延胡索酸水平显著降低。 与正常组相比，IL-6组的ROS水平显著增加。经过JPYQSX干预后，联合处理组的ROS水平显著降低。