Protective effect and mechanism of basic fibroblast growth factor on human chondrocyte injury induced by tumor necrosis factor-α combined with cycloheximide#br#
XIONG Chenghao1 YANG Bo1 TANG Daoqi1 XUE Qi2 FANG Haizhou1 WANG Jufang3
1.Zhuhai Essex Biopharmaceutical Co., Ltd, Guangdong Province, Zhuhai 519085, China;
2.Essex Bio-Technology Co., Ltd, Hangkong 999077, China;
3.School of Biological Science and Engineering, South China University of Technology, Guangdong Province, Guangzhou 510006, China
Abstract：Objective To investigate the protective effect and related mechanisms of basic fibroblast growth factor (bFGF) on human chondrocytes in an inflammatory environment in vitro. Methods Primary human chondrocyte injury model was established using 50 ng/ml tumor necrosis factor-α (TNF-α) combined with 1 μmol/L cycloheximide (CHX) and the effects of bFGF on the injury model were assessed. Low and high doses of bFGF groups were administered extra 1, 10 ng/ml bFGF based on model group, respectively. The effects of bFGF on the viability of model cells were detected by MTT assay after treatment for 48 hours, the effects of bFGF on apoptosis and mitochondrial membrane potential were investigated by flow cytometry after after 24 hours treatment, while the effects of bFGF on the expression of apoptosis-related proteins in model cells were detected by Western bolt after treatment for 24 hours. Results The cell viability of the model group (1 μmol/L CHX+50 ng/ml TNF-α) was significantly lower than that of the control group, and the difference was highly statistically significant (P ＜ 0.01); the cell viability of bFGF low-dose and high-dose groups were higher than that of the model group, and the differences were statistically significant (P ＜ 0.05). Flow cytometry results showed that the apoptosis rate of bFGF low-dose and high-dose groups was lower than that of the model group, and the difference was statistically significant (P ＜ 0.05). The results of JC-1 staining showed that the positive rate of FITC cells in the bFGF high-dose group was lower than that in the model group, and the difference was highly statistically significant (P ＜ 0.01). Western bolt showed that the expression level of Bcl-2 protein in the bFGF administration group was higher than that in the model group, while the expression level of Bax protein was lower than that in the model group, and the differences were statistically significant (P ＜ 0.05). Conlusion bFGF can protect chondrocytes from damage in inflammatory environment by inhibiting mitochondrial membrane damage and regulating the expression of apoptosis-related proteins Bax and Bcl-2, which has potential application prospects.
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