We found that the activation of MAPK and NF-κB signaling pathways and the nuclear translocation of phosphorylated p65 and p38 were inhibited prominently with the treatment of engeletin which may be the potential molecular mechanism for its anti-inflammation effects. Cell apoptosis was also attenuated with the addition of engeletin. Pretreatment with engeletin effectively ameliorate the TNF-α-induced up-regulation of inflammatory mediators and MMP families, promoting the anabolism of ECM meanwhile.

Results: Cell viability tests demonstrated there was little cytotoxicity of engeletin toward NP cells. The therapeutic effects of engeletin were detected through imaging and histology analysis. In vivo, percutaneous needle puncture was used to establish the IDD model in rat, and engeletin was administrated via intradiscal injection. The activation of NF-κB and MAPK pathways was evaluated by Western blotting and confocal microscopy. The expression of apoptosis-related proteins was tested by Western blotting. Cell apoptosis was analyzed by flow cytometry and TUNEL assay.

Using the TNF-α to simulate the inflammation status in vitro, the expression of inflammatory mediators and MMP families were determined by qPCR, Western blotting and confocal microscopy. Methods: The cytotoxicity of engeletin was validated by CCK-8 tests. Therefore, we aim to confirm its protective effects against the intervertebral disc degeneration in vivo and in vitro. Engeletin is a plant-derived medicine with anti-inflammation and antioxidant functions. Purpose: Low back pain (LBP) induced by intervertebral disc degeneration (IDD) brings progressively painful status and impairs the normal daily living. *These authors contributed equally to this workĬorrespondence: Tangjun Zhou Jie Zhao, Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200011, People’s Republic of China, Tel +8621-23271159, Fax +8621-63139920, Email Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200011, People’s Republic of China Baixing Li, * Xiao Yang, * Pu Zhang, Jiadong Guo, Kewei Rong, Xin Wang, Xiankun Cao, Tangjun Zhou, Jie Zhao

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