补肾活血方通过TGF-β/Glut1信号轴调控低氧环境下关节软骨细胞能量代谢稳态的作用机制研究

Osteoarthritis (OA) is a chronic degenerative disease. It has been found that the instability of energy metabolism of chondrocytes is the key cause of articular cartilage degeneration. Normal articular cartilage is in a hypoxic environment without blood vessels, and the energy of chondrocytes is mainly produced by glycolysis and supplemented by mitochondrial oxidative phosphorylation, which coordinate with each other to maintain the dynamic balance of energy metabolism. Previous studies showed that OA chondrocytes had abnormal energy metabolism. Activation of TGF-β/Smad2/3 in OA chondrocytes could increase Glut1 expression, promote glucose transport and glycolytic metabolism, maintain oxidative phosphorylation and restore the homeostasis of energy metabolism of cells. Bushen Huoxue recipe could reverse the degeneration of articular cartilage of OA through targeted TGF-β signaling pathway, and its effective monomer could regulate glycolytic metabolism of chondrocytes under hypoxia environment. Therefore, we propose a hypothesis that Bushen Huoxue recipe can regulate the homeostasis of energy metabolism of chondrocytes by regulating TGF-β/Glut1 signal axis under hypoxia environment, inhibit cartilage degeneration and matrix degradation, and play a role in the prevention and treatment of OA. In this project, transgenic knock-out technique and adenovirus transfection technique were used to elucidate the mechanism of Bushen Huoxue recipe in regulating the homeostasis of energy metabolism in chondrocytes in vitro and in vivo studies, providing a new target for the prevention and treatment of OA of traditional Chinese medicine.

骨性关节炎（OA）是一种慢性退行性疾病，研究发现软骨细胞能量代谢失稳是引起关节软骨退变的关键原因。正常关节软骨无血管处于低氧环境，软骨细胞能量来源于糖酵解（为主）与线粒体氧化磷酸化（为辅），二者相互协调维持能量代谢动态平衡。前期研究发现OA软骨细胞能量代谢异常，激活OA软骨细胞TGF-β/Smad2/3可以升高Glut1表达促进葡萄糖转运与糖酵解代谢，维持氧化磷酸化，恢复细胞能量代谢稳态；补肾活血方能通过靶向TGF-β信号通路逆转OA关节软骨退变，其有效单体对低氧环境下软骨细胞的糖酵解代谢具有调节作用。因此我们提出假说：补肾活血方能在低氧环境下通过调节TGF-β/Glut1信号轴来调控软骨细胞能量代谢稳态，抑制软骨退变及基质降解，起到防治OA的作用。本项目借助转基因敲基因技术与腺病毒转染技术在体内外联合阐明补肾活血方调控软骨细胞能量代谢稳态的作用机制，为中医药防治OA提供新靶点。

骨性关节炎（OA）是一种慢性退行性疾病，软骨细胞能量代谢失衡引起的关节软骨退变构成了OA形成的重要机制。正常关节软骨处于低氧环境，软骨细胞能量来源于糖酵解（为主）与线粒体氧化磷酸化（为辅），二者相互协调维持能量代谢动态平衡。前期研究发现OA软骨细胞出现了能量代谢稳态失衡现象，激活OA软骨细胞TGF-β信号可以调控能量代谢平衡，补肾活血方能通过靶向TGF-β信号通路逆转OA关节软骨退变，其有效单体对低氧环境下软骨细胞的糖酵解代谢具有调节作用。本项目研究明确了TGF-β/Smad2信号通过Glut1调控软骨细胞糖酵解代谢途径与线粒体氧化磷酸化途径、维持细胞能量代谢稳态的分子机制；以及补肾活血方在低氧环境下通过TGF-β/Glut1信号轴调控软骨细胞能量代谢稳态，抑制软骨细胞肥大分化、凋亡与ECM降解，从而延缓OA进程的作用机制。本项目借助内侧半月板失稳手术构建的OA小鼠模型，结合软骨细胞条件性基因敲除技术，通过体内外实验研究补肾活血方对OA关节软骨细胞的保护机制，为补肾活血方的临床推广应用提供科学依据。