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Chen S.-Q.,Fujian University of Traditional Chinese Medicine | Chen S.-Q.,Key Laboratory of Orthopedics and Exercise Rehabilitation of Ministry of Education | Lin J.-P.,Fujian University of Traditional Chinese Medicine | Lin J.-P.,Fujian Rehabilitation Laboratory of Motor Function | And 4 more authors.
Chinese Journal of Tissue Engineering Research | Year: 2013

BACKGROUND: Chinese herbaceous peony has been usually used in the treatment of cervical spondylosis in a frequency of over 30%. Paeoniflorin, known as one of the main monomers, has cell-protective abilities against inflammation and apoptosis. Despite these compelling observations, whether paeoniflorin protects annulus fibrosus cells is not reported. OBJECTIVE: To explore the proliferative and protective effects of paeoniflorin on intervertebral disc annulus fibrosus cells. METHODS: Annulus fibrosus cells derived from the intervertebral disc were digested with trypsin and collagenase type for the further culture in vitro. The third-passage annulus fibrosus cells were selected to establish an apoptosis model in response to Fas ligand (20 μg/L) and were interfered with paeoniflorin. The appropriate concentration of paeoniflorin was detected by the method of MTT. Apoptotic rate was evaluated with Annexin V/FITC double staining. RESULTS AND CONCLUSION: The 20. 8 and 2. 08 μmol/L paeoniflorin significantly promoted the proliferation of annulus fibrosus cells. The apoptotic rate of annulus fibrosus cells in vitro was significantly increased after induction with the 20 μg/L Fas ligand. 20. 8 μmol/L paeoniflorin could protect annulus fibrosus cells against Fas ligand-induced apoptosis.


Chen S.-Q.,Fujian University of Traditional Chinese Medicine | Chen S.-Q.,Key Laboratory of Orthopedics and Exercise Rehabilitation of Ministry of Education | Lin J.-P.,Fujian University of Traditional Chinese Medicine | Lin J.-P.,Fujian Rehabilitation Laboratory of Motor Function | And 5 more authors.
Chinese Journal of Tissue Engineering Research | Year: 2013

Background: Puerarin restrains the apoptosis of disc annulus fibrosus cells and delays the degenerationof the intervertebral disc. Objective: To explore the protective role of puerarin in rat cervical intervertebral disc annulus fibrosus cells. Methods: Cervical intervertebral disc annulus fibrosus cells were induced by different concentrations of Fas ligand (5, 20, 50 μg/L). The appropriate concentration of puerarin was detected using Annexin V-FITC-PI double staining to establish an in vitro apoptotic model of annulus fibrosus cells. Changes of annulus fibrosus cells in response to different concentrations of puerarin (0.01, 0.1, 1 g/L) were observed by flow cytometry and MTT method. Results and Conclusion: 5 μg/L Fas ligand had no significant influence on the survival rate and early-apoptotic rate of annulus fibrosus cells (P > 0.05). 20 and 50 μg/L Fas ligand significantly promoted annulus fibrosus cells apoptosis (P < 0.01) in a dose-dependent manner. The 0.01, 0.1 and 1 g/L puerarin could improve the survival rate of Fas ligand-induced annulus fibrosus cells in a dose-dependent manner (P < 0.01). These findings indicate that (1) Fas ligand can induce the disc annulus fibrosus to be in apoptosis status. (2) Puerarin (0.01, 0.1, 1 g/L) can promote the proliferation of disc annulus fibrosus cells in a dose-dependent manner. (3) The apoptosis rate of disc annulus fibrosus cells induced by Fas ligand in vitro can be restrained by puerarin interference.

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