椎间盘髓核细胞体外凋亡模型,动物造模 z-bio 2021-11-01 319

　　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 disc. Screening the highest pro-apoptotic concentration is used to detect the apoptotic rate of nucleus pulposus cells, the expression of apoptosis-related proteins Bax, bcl-2 and caspase-3 enzymes, cell proliferation curves 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. In the -3 enzyme 3% FBS group, the expression of bcl-2 was significantly higher than that of the 10% FBS group, and the expression of bcl-2 was reduced. The results of the CCK-8 study showed that in a medium containing 3%, FBS increased the proliferation rate of nucleus pulposus cells with the increase of the apoptosis rate. Its speed is slower. 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.