动物造模,椎间盘髓核细胞体外凋亡模型 z-bio 2021-04-13 262

　　Objective: To establish an in vitro apoptosis model of rat intervertebral disc nucleus pulposus cells.

　　Method: In order to fully simulate the microenvironment of degenerative disc nutrient deficiency, this experiment used DMEM medium containing 1%, 3%, 5%, 8% and 10% fetal bovine serum, and cultured the nucleus pulposus cells of the intervertebral disc. The optimal pro-apoptotic concentration was used to detect the apoptotic rate of nucleus pulposus cells, the expression of apoptosis-related proteins Bax, bcl-2 and caspase-3, cell proliferation curve and immunofluorescence analysis.

　　Result: The apoptosis rate of nucleus pulposus cells measured by flow cytometry increased with the decrease of fetal bovine serum (FBS) concentration, and 3% FBS was the most effective concentration for inducing apoptosis. Western blot showed Bax and caspase expression. -3 The expression of bcl-2 in the 3% FBS group was significantly higher than that in the 10% FBS group. The results of the CCK-8 study show that the medium containing 3% FBS has a higher rate of apoptosis. Increase the proliferation rate of nucleus pulposus cells. late. Immunofluorescence analysis showed that FAS expression in 3% FBS group was significantly higher than that in 10% FBS group.

　　Conclusion: 3% FBS induces nucleus pulposus cell apoptosis, which may eventually lead to the loss of cell function.