Department of Pharmacognosy, School of Pharmacy, Xinjiang Medical University, Urumqi, China; Department of Toxicology, School of Public Health, Xinjiang Medical University, Urumqi, China.
Department of Pharmacy, People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang, China.
Biomed Pharmacother. 2022 Jun;150:113004. doi: 10.1016/j.biopha.2022.113004. Epub 2022 Apr 28.
To study whether ACT exerts anti-fatigue activity against CRF by inducing skeletal muscle mitophagy via suppressing PHD2 to upregulate the HIF-1α/BNIP3 signaling pathway.
In this study, the molecular docking virtual screening technique was used to screen active components in Cistanche tubulosa that act as potential PHD2 inhibitors; the preliminary verification was carried out by Surface plasmon resonance (SPR) technology. BALB/c mice were treated with Paclitaxel (PTX, 10 mg/kg) and ACT (50, 100 mg/kg) alone or in combination for 20 days. Fatigue-related behaviors, energy metabolism and skeletal muscle mitochondria were assessed. Murine C2C12 myoblast was cultured and differentiated; then, a C26 tumor cell-conditioned medium was added to induce cachexia. Intracellular reactive oxygen species (ROS), mitochondrial membrane potential, mitochondrial microstructure and function, autophagy, PHD2/HIF-1 and PINK1/Parkin signal pathway proteins were analyzed. Then, interfering RNA technology was used to silence PHD2 and observe the efficacy of ACT.
We demonstrated that ACT exerted good binding activity with PHD2; ACT administration ameliorated PTX-induced muscle fatigue-like behavior via improving muscle quality and mitochondria function, increasing mitophagy, upregulating COXIV, CytoC, PINK1, Parkin, HIF-1α and BNIP3 expression and inhibiting p62, LC3B, PHD2 and Beclin-1 expression. The protective effect of ACT disappeared after transfection with the PHD2 gene knockdown plasmid Egln-1-RNAi.
These results suggest that ACT can improve CRF by promoting mitophagy via suppression of PHD2 to remove dysfunctional mitochondria, demonstrating that ACT has huge prospects for clinical application in CRF treatment.
通过抑制 PHD2 来上调 HIF-1α/BNIP3 信号通路，研究 ACT 是否通过诱导骨骼肌自噬来发挥抗 CRF 疲劳作用。
在这项研究中，使用分子对接虚拟筛选技术筛选作为潜在 PHD2 抑制剂的肉苁蓉中的活性成分；通过表面等离子体共振（SPR）技术进行初步验证。BALB/c 小鼠单独或联合使用紫杉醇（PTX，10mg/kg）和 ACT（50、100mg/kg）治疗 20 天。评估疲劳相关行为、能量代谢和骨骼肌线粒体。培养和分化鼠 C2C12 成肌细胞；然后加入 C26 肿瘤细胞条件培养基诱导恶病质。分析细胞内活性氧（ROS）、线粒体膜电位、线粒体微观结构和功能、自噬、PHD2/HIF-1 和 PINK1/Parkin 信号通路蛋白。然后，使用干扰 RNA 技术沉默 PHD2 并观察 ACT 的疗效。
我们证明 ACT 与 PHD2 具有良好的结合活性；ACT 通过改善肌肉质量和线粒体功能、增加自噬、上调 COXIV、CytoC、PINK1、Parkin、HIF-1α 和 BNIP3 表达以及抑制 p62、LC3B、PHD2 和 Beclin-1 表达，改善了 PTX 诱导的肌肉疲劳样行为。在用 PHD2 基因敲低质粒 Egln-1-RNAi 转染后，ACT 的保护作用消失。
这些结果表明，ACT 通过抑制 PHD2 促进自噬来改善 CRF，从而去除功能失调的线粒体，表明 ACT 在治疗 CRF 方面具有巨大的临床应用前景。
