胰岛素样生长因子-1对白介素-1体外诱导家兔椎间盘髓核细胞凋亡的影响  

Effect of insulin-like growth factor-1 on the apoptosis of rabbit nucleus pulposus cells induced by interleukin-1

作　　者：张长春[1] 周建生[1] 王祥[2] 周新社[1] 孙宝安[1] 邵晨[1] 林清华[1] 

机构地区：[1]蚌埠医学院第一附属医院,安徽省组织移植重点实验室,蚌埠233004 [2]浙江大学医学院第一附属医院,杭州310000

出　　处：《解剖学杂志》2013年第2期152-155,共4页Chinese Journal of Anatomy

基　　金：安徽省高校省级自然科学研究重点项目（KJ2011A204）

摘　　要：目的：利用白介素-1（IL-1）诱导培养的家兔椎间盘髓核细胞凋亡，用胰岛素样生长因子-1（IGF-1）对凋亡进行干预，探讨IGF-1对髓核细胞凋亡的抑制作用。方法：分离健康家兔椎间盘髓核，Ⅱ型胶原酶消化法分离椎间盘髓核细胞。Ⅱ型胶原免疫组织化学显色和甲苯胺蓝染色对髓核细胞鉴定。培养第1代髓核细胞分为空白组、IGF-1组和IL-1组。利用Giemsa染色、原位末端标记（TUNEL）、流式细胞仪观测各组髓核细胞凋亡情况。结果：IL-1组可见大量髓核细胞凋亡，IGF-1组可见少量细胞凋亡，空白组罕见凋亡征象。原位末端标记显示，与IL-1组比较，IGF-1组细胞凋亡率明显降低。流式细胞仪检测显示IGF-1组细胞凋亡率明显降低。IL-1组与空白组比较，凋亡率明显增高。IGF-1组、IL-1组和空白组晚期凋亡百分率差异均有统计学意义。结论：IGF-1对IL-1体外诱导的髓核细胞凋亡具有抑制作用。Objective： To investigate the effects of insulin-like growth factor-1 （IGF-1） on the apoptosis of rabbits nucleus pul posus cells induced by interleukin-1（IL-1）. Methods： The nucleus pulposus was removed from intervertebral discs of healthy domestic rabbits. The cells isolated from these structures were cultured to a monolayer, and then digested with collagenase Ⅱ and purified using trypsin and differential adhesion methods. Nucleus pulposus cells were then identified using type Ⅱcollagen immunohistochemistry and were stained with toluidine blue. Following exposure to IGF1 or IL-1, they were observed by light microscopy. Giemsa staining, TdT-mediated dUTP-biotin nick end labeling （TUNEL）, and flow cytometry were used to de- tect the rate of early cell death, which served as an indicator of apoptosis. Results： A large number of nucleus pulposus cells underwent apoptosis. Morphological changes typically associated with apoptosis were observed in the IL-1 group without obvi- ous signs of apoptosis. A small but significant degree of apoptosis was seen in the IGF-1 group. TUNEL revealed that the rate of apoptosis in the IGF-1 group was significantly reduced compared with the IL-1 group, which was confirmed using flow cy tometry. Compared with the control group, the rate of apoptosis was also significantly increased in IL-1 exposed cells. Conclusion： These findings suggest that IGF1 inhibits the apoptosis of nucleus pulposus cells induced by IL-1.

关 键 词：椎间盘退变 髓核 凋亡 胰岛素样生长因子-1 白介素-1 原位末端标记 

分 类 号：R587.2[医药卫生—内分泌]