摘要
目的探讨不同浓度及时间的尼古丁在人椎间盘髓核细胞退变中的作用。方法随机选择5例腰椎骨折患者的椎间盘髓核组织进行体外细胞培养,种于12孔板中,按照0、1、33.3、100、200、500ng/m L的尼古丁浓度以及1、2、3、7、10天加入来培养,每组细胞用Trizol法提取总RNA,用荧光定量PCR测定AQP1,AQP3和II型胶原(Collagen)以及蛋白多糖(Aggrecan)的m RNA表达。用SPSS19.0进行统计分析。结果随着尼古丁浓度升高及培养时间的延长,髓核细胞形态变化明显,贴壁能力减弱,胞质减少,胞核萎缩,空泡形成,凋亡增加。且细胞AQP1及AQP3的m RNA水平随着尼古丁浓度及时间的增加而降低,II型胶原及蛋白聚糖m RNA水平也降低,细胞发生退变。结论尼古丁会降低人椎间盘髓核细胞AQP1和AQP3的表达,进而降低髓核细胞II型胶原和蛋白聚糖的表达,促进椎间盘退变,且作用时间越长,浓度越高,细胞退变越严重。
Objective To evaluate the effect of nicotine on the degeneration of human nucleus pulposus cells. Methods 5 patients undergoing lumbar discectomy surgeries in our department were randomly included. The nucleus pulposus tissues were cultured in well plate and the concentrations of nicotine were 0, 1, 33.3, 100, 200, 500ng/mL. After 1, 2, 3, 7, 10 days, RNA of cells were extracted using Trizol. The mRNA expression ofAQP1, AQP3, collagen II and Aggrecan were valued by rtPCR. Data analysis was done by SPSS19.0. Results With an increase in nicotine concentration and culture time, the morphology of nucleus pulposus changed significantly, weakening of the adherent ability, showing decrease of cytoplasm, atrophy of cell nucleus and increase ofapotosis. The mRNA expression ofAQP 1 and AQP3 also decreased and so did collagen II and aggrecan. Conclusion Nicotine can decrease the mRNA expression of AQP land AQP3 of nucleus pulposus, thus, causing a decrease in the expression of type II collagen and aggrecanto ac- celerate disc degeneration. This effect seems to be time and dose dependent.
作者
黄宇峰
雍之瑶
刘晓明
李浩曦
王善金
巴兆玉
沈彬
赵卫东
吴德升
Huang Yufeng Yong Zhiyao LiuXiaoming et al(Department of Spine Surgery, Shanghai East Hospital, Shanghai, 200120, China)
出处
《生物骨科材料与临床研究》
CAS
2017年第1期16-19,I0006,I0007,共6页
Orthopaedic Biomechanics Materials and Clinical Study
基金
国家自然科学基金面上项目(81672199)
上海市教委产学研项目课题(15cxy50)
2013年上海市东方医院朝阳人才计划(DFZY-8)
浦东新区卫生系统重点学科建设资助(PWzx2014-02)
关键词
尼古丁
椎间盘
髓核细胞
退变
水通道蛋白
Nicotine
Intervertebral disc
Nucleus pulposus cell
Degeneration
Aquaporins
