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白细胞介素-1β诱导人椎间盘髓核细胞凋亡(1)
http://www.100md.com 2011年7月25日 王新光 叶 伟 郭汉明 康 明 陈秋兰 杜开利 黄东生
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     [摘要] 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20 μg/L和200 μg/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 20 μg/L组中细胞凋亡率(8.46%)和IL-1β 200 μg/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 200 μg/L组是IL-1β 20 μg/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。

    [关键词] 椎间盘退变; 髓核细胞; 细胞培养;白细胞介素-1β;凋亡

    [中图分类号] R681.5[文献标识码]A [文章编号]1673-7210(2011)07(c)-020-05

    IL-1 β induce apoptosis of human nucleus pulposus cells

    WANG Xinguang1, YE Wei2, GUO Hanming1, KANG Ming1, CHEN Qiulan3, DU Kaili2, HUANG Dongsheng2*

    1.Department of Orthopedics, Huizhou Municipal Central Hospital, Guangdong Province, Huizhou 516001, China; 2.Department of Orthopedics, the Second Affiliated Hospital of Sun Yat-sen University, Guangdong Province, Guangzhou 510120,China; 3.Department of Ultrosound B, Huizhou Municipal Central Hospital, Guangdong Province, Huizhou 516001, China

    [Abstract] Objective: To explore the culture of human normal nucleus pulposus cells and evaluate the effect of IL-1β 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 O 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 by trypsin and collagenase and cultured by Dulbecco′sModified Eagle Medium and Ham′s F-12 medium. The expression of glycosaminoglycan, proteoglycan and collagen Ⅱ could be detected in human nucleus pulposus cells ......

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