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Danlu Tongdu tablets treat lumbar spinal stenosis through reducing reactive oxygen species and apoptosis by regulating CDK2/CDK4/CDKN1A expression

Lumbar spinal stenosis (LSS) is caused by the compression of the nerve root or cauda equina nerve by stenosis of the lumbar spinal canal or intervertebral foramen, and is manifested as chronic low back and leg pain. Danlu Tongdu (DLTD) tablets can relieve chronic pain caused by LSS, but the molecular mechanism remains largely unknown. In this study, the potential molecular mechanism of DLTD tablets in the treatment of LSS was firstly predicted by network pharmacology method. Results showed that DLTD functions in regulating anti-oxidative, apoptosis, and inflammation signaling pathways. Furthermore, the flow cytometry results showed that DLTD tablets efficiently reduced ROS content and inhibited rat neural stem cell apoptosis induced by hydrogen peroxide. DLTD also inhibited the mitochondrial membrane potential damage induced by hydrogen peroxide. Elisa analysis showed that DLTD induced cell cycle related protein, CDK2 and CDK4 and reduced CDKN1A protein expression level. Taken together, our study provided new insights of DLTD in treating LSS through reducing ROS content, decreasing apoptosis by inhibiting CDKN1A and promoting CDK2 and CDK4 expression levels.