外源性心脏营养素1:保护PC12细胞和许旺细胞的可能性  

Exogenous cardiotrophin-1: The possibility to protect PC12 cells and Schwann cells

作　　者：辛泽团 刘湘泽[2] 张明进[2] 李伟[2] 陈伯华[2] 马学晓[2] 

机构地区：[1]青岛市城阳区人民医院骨科,山东省青岛市266100 [2]青岛大学医学院附属医院脊柱外科,山东省青岛市266003

出　　处：《中国组织工程研究》2013年第41期7265-7271,共7页Chinese Journal of Tissue Engineering Research

基　　金：山东省自然科学基金(Y2008C41)资助~~

摘　　要：背景:外周神经损伤后的很短时间内,神经和肌肉已经开始发生不可逆的退变。如何延缓外周神经损伤后的退变过程,并创造一定的微环境作为其恢复的基础。目的:探讨外源性心脏营养素1保护PC12细胞和许旺细胞的作用。方法:获得并培养许旺细胞和PC12细胞,分别进行完全培养基培养、无血清培养基培养和50℃高温培养,心脏营养素1组中加入一定量的外源性心脏营养素1溶液,对照组中加入等量无菌DMEM溶液培养24 h。应用CCK-8比色法测定PC12细胞和许旺细胞的存活率,乳酸脱氢酶试剂盒测定上清液中乳酸脱氢酶活性,硫代巴比妥酸法测定丙二醛浓度,黄嘌呤氧化酶法测定超氧化物歧化酶活性。结果与结论:加入心脏营养素1后PC12细胞和许旺细胞存活率明显增高,乳酸脱氢酶释放量、丙二醛浓度明显减少,超氧化物歧化酶活性显著增加。证实外源性心脏营养素1能够明显减轻缺血和热应激刺激对PC12细胞和许旺细胞的损伤从而起到保护作用,其作用机制可能与心脏营养素1与细胞表面受体结合激活了抗凋亡蛋白的表达有关。BACKGROUND： Restoration of neurological functions after the damaged peripheral nerve is reconstructed is a hot topic in existing research. Within a short term following peripheral nerve injury, nerve and muscle begin to develop irreversible degeneration. Restoration of the damaged nerve requires delayed degeneration and basic microenvironment. OBJECTIVE： To investigate the protective effect of cardiotrophin-1 on PC12 cells and Schwann cells. METHODS： Schwann cells and PC12 cells were obtained and cultured in complete medium, serum-free medium and 50 12 medium, respectively. Cells in cardiotrophin-1 group were treated with exogenous cardiotrophin-1 solvent, while those in the control group were treated with equivalent Dulbecco＇s modified Eagle＇s medium, for 24 hours. The survival rate for PC12 cells and Schwann cells was determined using Cell Counting Kit-8 colorimetric method. The lactate dehydrogenase activity in supernatant was detected by lactate dehydrogenase kit, and the ma!ondialdehyde content and superoxide dismutase activity were measured by thiobarbituricaicd and xanthine oxidese method respectively. RESULTS AND CONCLUSION： The survival rate of PC12 cells and Schwann cells in cardiotrophin-1 group was obviously increased, lactate dehydregenase releasing and malondialdehyde content were obviously decreased, superoxide dismutase activity was dramatically improved compared with control group. Exogenous cardiotrophin-1reduces the injury caused by ischemia and heat stress stimulation for PC12 cells and Schwann ceils. The mechanism of protection may be related to the expression of anti-apoptosis protein activated by the combination of cardiotrophin-1 and its receptors.

关 键 词：组织构建 组织构建细胞学实验 生物活性因子 心脏营养素1 PC12细胞 许旺细胞 CCK-8 乳酸脱氢酶试剂盒 硫代巴比妥酸法 黄嘌呤氧化酶法 保护作用 省级基金 

分 类 号：R318[医药卫生—生物医学工程]