Objective: To explore the culture of human normal nucleus pulposus cells and evaluate the effect of IL-1 β on the apoptosis of human nucleus pulposus cells. Methods: Human normal nucleus pulposus tissue were separated from the idiopathic scoliosis patients, and the nucleus pulposus cells were isolated with sequential trypsin and collagenase, the expression of glycosaminoglycan, proteoglycan and collagen Ⅱ were detected by the toluidine blue, safranin 0 and immunohistochemistry of collagen Ⅱ. Then stimulated with 20 μg/L and 200 μg/L human recombinated IL-1β for 24 hours, the apoptosis rate was detected. Results: The human nucleus pulposus cells could be isolated hy trypsin and collagenase and cultured by Dulbecco's Modified Eagle Medium and Ham's F-12 medium. The expression of glycosaminoglycan, proteoglycan and collagen Ⅱ could be detected in human nucleus pulposus cells. The apoptosis rate in IL-1β 20 μg/L group (8.46％) and IL-1 β 200 μg/L group (19％) was 2.95 times and 6.63 times of the control group (2.86％) respectively (P＜0.05),IL-1β 200 μg/L group was 2.24 times of IL-1β 20 μg/L group (P＜0.05). Conclusion: The nucleus pulposus cells can be isolated and cuhured with trypsin and collagenase, IL-1 β can induce the apoptosis of human nucleus pulposus cells. IL-1β plays an important role in the pathogenesis of intervertebral disc degeneration.%目的:探讨人正常椎间盘髓核细胞的培养方法以及白细胞介素-1β(IL-1β)对人椎间盘髓核细胞凋亡的作用.方法:取特发性脊柱侧弯患者腰椎间盘髓核组织,用胰酶、胶原酶序贯消化法消化细胞,并用甲苯胺蓝、番红-O染色和Ⅱ型胶原免疫组化分别检测糖胺多糖、蛋白多糖和Ⅱ型胶原表达,分别用20μg/L和200μg/L重组人IL-1β刺激髓核细胞24 h,检测其凋亡率.结果:用酶消化法可成功分离培养人椎间盘髓核细胞,检测所培养细胞可表达糖胺多糖、蛋白多糖和Ⅱ型胶原,IL-1β 20μg/L组中细胞凋亡率(8.46%)和IL-1β 200μg/L组(19.00%)分别为空白对照组(2.86%)的2.95倍和6.63倍(P<0.05),IL-1β 200μg/L组是IL-1β 20μg/L组的2.24倍(P<0.05).结论:用酶消化法可成功培养人椎间盘髓核细胞,IL-1β可以诱导椎间盘髓核细胞凋亡,并且随着浓度升高凋亡率增加,提示炎症因子IL-1β在退变椎间盘细胞凋亡中起了重要的作用.
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