生长因子对椎间盘终板软骨细胞抗氧化能力的影响

Effects of different growth factors on the antioxidant capacity of endplate chondrocytes

背景:维生素E的抗氧化能力众所周知,但其他生长因子是否对终板软骨细胞具有抗氧化能力,国内外未见报道。目的:观察不同生长因子对椎间盘终板软骨细胞抗氧化能力的影响。方法:将原代培养获取的终板软骨细胞后分为4组,即空白对照组、血清剥夺组、过氧化氢刺激组、过氧化氢刺激后+不同生长因子干预组。第4组再细分为胰岛素生长因子1组、碱性成纤维细胞生长因子组、转化生长因子β组、Forskolin组、维生素E组,共5个亚组,通过RT-PCR检测各组半胱氨酸蛋白酶3、基质金属蛋白酶13、基质金属蛋白酶3、基质金属蛋白酶抑制剂1及凝血酶4、凝血酶5等基因表达情况;通过凋亡试剂盒及流式细胞仪分析不同组内细胞凋亡情况;Western blot检测细胞的合成与分泌;使用试剂盒检测总抗氧化能力和过氧化氢含量,并进行统计学分析。结果与结论:(1)不同生长因子对终板软骨细胞凋亡、分泌和抗氧化能力的影响存在差异;(2)总体来讲,转化生长因子β和Forskolin对过氧化氢刺激后的细胞具有进一步损伤作用,而胰岛素生长因子1和维生素E对损伤后的细胞存在保护作用。

BACKGROUND： It is well-known that vitamin E holds antioxidant capacity, but whether other growth factors have the same effect on endplate chondrocytes has not yet been reported. OBJECTIVE： To observe the effect of different growth factors on the antioxidant ability of endplate chondrocytes in the intervertebral disc. METHODS： Endplate chondrocytes were primary cultured, and then divided into four groups, including blank control, serum deprivation, hydrogen peroxide stimulation and hydrogen peroxide stimulation combined with different growth factors groups. The 4th group was subdivided into insulin-like growth factor-1, basic fibroblast growth factor, transforming growth factor β, forskolin and vitamin E groups. The expression levels of caspase-3, matrix metalloproteinase 13 and 3, inhibitor of metalloproteinase 1 as well as thrombin 4 and 5 were detected by real-time PCR. Cell apoptosis was analyzed through apoptosis kit and flow cytometry. Cell synthesis and secretion were detected by western blot assay. The total antioxidant capacity and the hydrogen peroxide content were determined by kit, and then statistically analyzed. RESULTS AND CONCLUSION： Different growth factors had significant differences in the endplate chondrocyte apoptosis, secretion and antioxidant capacity. To conclude, transforming growth factor β and forskolin do further damage to the cells stimulated by hydrogen peroxide, while insulin-like growth factor-1 and vitamin E expose protective effect on the injured cells.