葡萄糖对体外培养椎间盘髓核细胞的影响  

Effect of glucose on nucleus pulposus cells cultured in vitro

作　　者：张世磊[1] 梁伟[1] 杨勇[1] 杜俊杰[1] 罗卓荆[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,陕西西安710032

出　　处：《现代生物医学进展》2010年第24期4647-4649,共3页Progress in Modern Biomedicine

基　　金：国家自然科学基金资助(81000813)

摘　　要：目的:探讨葡萄糖对体外培养髓核细胞的生物学特性的影响。方法:酶消化法分离培养正常椎间盘髓核细胞。对照组:DF12+20%FBS培养液(葡萄糖浓度1000mg/L)、无糖组:无糖DMEM+20%FBS(葡萄糖浓度0mg/L)培养液培养髓核细胞。HE染色观察细胞形态变化,计数板计数细胞总数,台盼蓝染色计算髓核细胞活性比率,流式细胞仪检测细胞凋亡率,Hoechst33258染色观察凋亡细胞核的变化。结果:两组培养液培养细胞形态大体正常,并无明显变化。对照组细胞总数明显多于无糖组。细胞活性率对照组也高于无糖组。Hoechst33258染色凋亡细胞,凋亡细胞核内可见致密的颗粒状和块状荧光,细胞核形态不规则,少数细胞核碎裂,部分细胞核呈月牙形。结论:葡萄糖对椎间盘髓核细胞的增殖及凋亡有显著的影响。Objective： To investigate the influence of glucose on the biological characteristics of nucleus pulposus cells in vitro.Methods： The normal human nucleus pulposus cells were isolated and cultured by collagenase digestion methods.The normal human nu-cleus pulposus cells were cultured in culture solutions of DF12＋20%FBS（the density of glucose was 1000mg/L） for control group,DMEM＋20%FBS（the density of glucose was 0mg/L） for glucose deprivation group.Morphologic changes of the cells were detected by HE staining.The number of nucleus pulposus cells was counted by counting plate.The cell activity rates were detected by trypan blues staining.The cell apopototic rates were detected by flow cytometry.The change of apoptotic nucleus pulposus cells nucleus were exam-ined by Hoechst33258 staining.Results： The cellular morphology was normal in glucose deprivation group and control group in vitro.The total number of in control groups was more than that in glucose deprivation groups.The cellular activity in control groups were more than that in glucose deprivation groups.Hoechst33258 staining showed that there were condensed or broken nucleus and the number of apoptotic cells increased in glucose deprivation groups.Conclusion： Glucose has a significant effect to the proliferation and apoptosis of disc nucleus pulposus cells.

关 键 词：葡萄糖 髓核细胞 退变 

分 类 号：R681.53[医药卫生—骨科学]