芹菜素通过抗炎性反应和细胞凋亡保护大鼠髓核细胞

The anti-imflammation and anti-apoptosis effects of apigenin on the nucleus pulposus cells of rats

目的探讨芹菜素(API)对大鼠髓核细胞(NPs)的保护作用及相关机制。方法取第3代髓核细胞经100μM过氧化氢(H_2O_2)处理2 h制作大鼠髓核细胞损伤模型,实验分为5组:对照组,模型组、API 10μM组、API 20μM组和API 40μM组,对照组为正常髓核细胞,模型组给与100μM H_2O_2处理2 h,实验组分别给与API 10μM、20μM和40μM处理,24 h后流式细胞仪检测细胞凋亡率;Western blot法检测凋亡蛋白Bcl-2和Bax表达;ELISA法检测肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)以及caspase-3、caspase-9表达。结果 24 h后,模型组细胞凋亡率显著高于对照组,而与模型组比较,芹菜素组能显著抑制髓核细胞的凋亡,并呈剂量依赖趋势(F=37.547,P=0.034);Western blot显示,芹菜素能显著提高Bcl-2表达,并呈剂量依赖趋势(F=22.016,P=0.009),抑制Bax蛋白表达(F=56.971,P=0.000);ELISA结果显示,芹菜素能明显抑制TNF-α、IL-1β以及caspase-3、caspase-9的表达,并呈剂量依赖趋势(F=11.753,P=0.018;F=39.467,P=0.004;F=14.185,P=0.021;F=9.663,P=0.037)。结论芹菜素对髓核细胞的保护作用可能是通过抑制炎性反应以及抗细胞凋亡作用完成,且其作用呈剂量依赖趋势。

Objective To explore the protective effect of apigenin on the nucleus pulposus cells and its potential mechanism. Methods Damage model of nucleus pulposus cells were established after pretreated with 100 μmol/ L H2 O2 . 5 groups were divede into normal group,model group,apigenin 10 μmol/ L,apigenin 20 μmol/ L and apigenin 40 μmol/ L group. After treatment for 24 h,cell apoptosis were detected by flow cytometry. Bcl - 2 and Bax protein expression were valued by western blot. TNF - α,IL - 1β,caspase - 3 and caspase - 9 activities were detec-ted by elisa. Results The apoptosis rates in model group were significantly higher than that in the control group,while apigenin markedly sup-press the apoptosis rates compared with the model group,which showed a dose - dependent trend （F = 37. 547,P = 0. 034）. Western blot showed that apigenin could effectively up regulate the Bcl - 2 protein expression,which showed a dose - dependent trend （F = 22. 016,P =0. 009）and down regulate the Bax expression （F = 56. 971,P = 0. 000）. ELISA showed that apigenin effectively suppressed the activities of TNF - α,IL - 1β,caspase - 3 and caspase - 9,compared with model group,which showed a dose - dependent trend （F = 11. 753,P = 0. 018;F = 39. 467,P = 0. 004;F = 14. 185,P = 0. 021;F = 9. 663,P = 0. 037）. Conclusion Apigenin protects the nucleus pulposus cells via its anti - imflammation and anti - apoptosis pathways,which showed a dose - dependent trend.