Acupotomy Activates PI3K/Akt Signaling Pathway Mediated by FGF7,10/KDR Axis to Regulate Apoptosis

Jia-Ming Qian,1 Xiao-Le Wang,1 Ting Fang,1 Lang Liu,1 Fu-Shui Liu,1,2 Xiao-Lan Zhao,2 Khaliunaa Tumurbaatar2,3 1Affiliated Hospital of Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, 330006, People’s Republic of China; 2Graduate School of Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, 330004, People’s Republic of China; 3Institute of Traditional Medicine and Technology, Ulan Bator, 17032, MongoliaCorrespondence: Fu-Shui Liu, Email [email protected]: Acupotomy in the treatment of cervical spondylosis (CS) is effective and widely used in clinic, but the mechanism is still unclear. In this study, we started from treating CS with acupotomy “regulating muscles to treat bone disorders” and cervical myocyte apoptosis, the mechanism of acupotomy activates PI3K/Akt signaling pathway mediated by FGF7,10/KDR signaling axis to regulate apoptosis was explored, providing theoretical basis for clarifying the target and molecular mechanism of acupotomy for CS.Methods: Seventy-four SPF SD rats were divided into 9 groups, including acupotomy group, sham operation group, model group, normal group, control AAV group, control AAV+model group, AAV+model group, and AAV+model+acupotomy group. The CS model was verified by cervical spine X-ray and transmission electron microscopy. Multiple immunofluorescence and COIP techniques were used to verify the interaction between FGF7, FGF10 and KDR proteins in splenius capitis. The tension and pain threshold of rats were measured to evaluate motor and sensory disorders. HE staining was used to observe the pathological changes of splenius capitis. Finally, the expressions of FGF7, FGF10, KDR, PI3K, Akt, bcl2 and Caspase9 were detected by qPCR and Western blot.Results: X-ray and transmission electron microscopy showed that the CS model was successfully constructed. Multiple immunofluorescence and COIP techniques identified the interaction between FGF7, FGF10 and KDR proteins. Acupotomy can effectively improve the tension and pain threshold of CS rats and also improve cervical vertebrae degeneration. After modeling, the expressions of FGF7, FGF10, KDR and Caspase9 were increased, while the expressions of PI3K, Akt and bcl2 were decreased, which could be reversed by acupotomy. The effect is alleviated after knockdown of KDR, and the downstream signal transmission is blocked.Conclusion: Acupotomy activates PI3K/Akt signaling pathway mediated by FGF7,10/KDR signaling axis, regulates cervical myocyte apoptosis, and repairs strained muscles, which may be the key molecular mechanism of treating CS with acupotomy “regulating muscles to treat bone disorders”. Keywords: acupotomy, cervical spondylosis, regulating muscles to treat bone disorders, FGF, KDR, apoptosis, adeno-associated virus