过氧化物酶Ⅱ对体外培养人退变腰椎间盘髓核细胞的抑制作用

背景:椎间盘退变可导致髓核细胞数量的减少,过氧化物酶Ⅱ可参与细胞的抗氧化损伤、细胞分裂、分化、信号转导和凋亡等过程的调控.过氧化物酶Ⅱ对椎间盘退变有促进作用,但其机制仍不清楚.目的:观察过氧化物酶Ⅱ对体外培养的人退变腰椎间盘髓核细胞的活性和Ⅱ型胶原合成的影响.方法:体外培养人退变腰椎间盘髓核细胞,设置对照组及过氧化物酶Ⅱ10,100和1 000 ng/L组.对照组不含氧化物酶Ⅱ,其他3组加入相应剂量的氧化物酶Ⅱ.应用免疫组化法鉴定髓核细胞,并采用cck-8试剂盒检测细胞增殖情况,于加过氧化物酶Ⅱ后第3和7天分别取对照组及各组细胞上清液,采用双抗体夹心酶联免疫吸附实验测定Ⅱ型胶原表达情况.结果与结论:在体外培养的人退变腰椎间盘髓核细胞,随加入的过氧化物酶Ⅱ质量浓度的增加,椎间盘髓核细胞数量和Ⅱ型胶原合成逐渐减少(P＜0 01).提示过氧化物酶Ⅱ对椎间盘髓核细胞的数量和Ⅱ型胶原合成有明显的抑制作用,并呈剂量依赖关系.以此推测过氧化物酶Ⅱ对髓核细胞的抑制作用可能是导致椎间盘退变的一种促发因素.

Inhibition of peroxiredoxin Ⅱ on human intervertebral disc cells cultured in vitro

BACKGROUND:intervertebral disc degeneration can reduce nucleus pulposus cells,and peroxiredoxin II involved in the regulation of resist oxidation damage,cell division,differentiation,signal transduction and apoptosis.Peroxiredoxin Ⅱ has promotive effect on intervertebral disc degeneration,whereas the mechanism remains poorly understood.OBJECTIVE:To observe the effects of perexiredoxin Ⅱ on human intevertebral disc cells activity and type Ⅱ collagen synthesis in vitro.METHODS:Human degenerated human lumbar disc cells were cultured in vitro,and assigned into the control and peroxidase Ⅱ groups.Peroxidase Ⅱ with doses of 10,100 and 1 000 ng/L were added into the peroxidase Ⅱ groups.The cells were identified by immunohistochemical staining,and the cell proliferation was detected using cck-8 kit.Cell supematant was collected at days 3 and 7 after operation,and the expression of type Ⅱ collagen was measured by double-antibody sandwich enzyme-linked immunosorbent assay.RESULTS AND CONCLUSION:In vitro cultured human degenerative lumbar intervertebral disc nucleus pulposus cells by adding peroxidase increased with the dose-Ⅱ,the disc nucleus pulposus cells of the volume and type Ⅱ collagen synthesis gradually reduced(P ＜ 0.01).Tips peroxidase Ⅱ on the intervertebral disc nucleus pulposus cells,the number and type Ⅱ collagen synthesis significantly inhibited in a dose-dependent manner.Thus speculated that peroxidase Ⅱ on the nucleus pulposus cells in vitro may lead to disc degeneration as a precipitating factor.