Puerarin (PUR), an 8-C-glucoside of daidzein extracted from Pueraria plants, is closely related to autophagy, reduced reactive\noxygen species (ROS) production, and anti-inflammatory effects, but its effects on human nucleus pulposus mesenchymal stem\ncells (NPMSCs) have not yet been identified. In this study, NPMSCs were cultured in a compression apparatus to simulate the\nmicroenvironment of the intervertebral disc under controlled pressure (1.0 MPa), and we found that cell viability was decreased\nand apoptosis level was gradually increased as compression duration was prolonged. After PUR administration, apoptosis level\nevaluated by flow cytometry and caspase-3 activity was remitted, and protein levels of Bas as well as cleaved caspase-3 were\ndecreased, while elevated Bcl-2 level was identified. Moreover, ATP production detection, ROS, and JC-1 fluorography as well as\nquantitative analysis suggested that PUR could attenuate intercellular ROS accumulation and mitochondrial dysfunction.\nBesides, the rat tail compression model was utilized, which indicated that PUR could restore impaired nucleus pulposus\ndegeneration induced by compression. The PI3K/Akt pathway was identified to be deactivated after compression stimulation by\nwestern blot, and PUR could rescue the phosphorylation of Akt, thus reactivating the pathway. The effects of PUR, such as\nantiapoptosis, cell viability restoration, antioxidation, and mitochondrial maintenance, were all counteracted by application of\nthe PI3K/Akt pathway inhibitor (LY294002). Summarily, PUR could alleviate compression-induced apoptosis and cell death of\nhuman NPMSCs in vitro as well as on the rat compression model and maintain intracellular homeostasis by stabilizing\nmitochondrial membrane potential and attenuating ROS accumulation through activating the PI3K/Akt pathway.
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