摘要
背景:椎间盘退变引起的下腰痛是导致老年人瘫痪的主要原因,但是有关椎间盘退变的机制仍未完全明确。自噬可能在椎间盘退变过程中起重要作用。目的:研究自噬对大鼠腰椎间盘软骨终板细胞Ⅱ型胶原纤维α1(COL2A1)和聚集蛋白聚糖(aggrecan)分泌的影响。方法:将细胞分为对照组、巴弗洛霉素(Baf)组、肿瘤坏死因子-α(TNF-α)组、TNF-α+Baf组4组,其中对照组、Baf组、TNF-α组分别采用二甲基亚砜、Baf、TNF-α作用于细胞24 h,TNF-α+Baf组先用Baf阻断自噬,再用TNF-α作用于细胞24 h。分别通过实时定量荧光聚合酶链反应、蛋白质印迹法、细胞免疫荧光染色等方法检测各组COL2A1、aggrecan mRNA和蛋白的表达并进行比较。结果:Baf组、TNF-α组和TNF-α+Baf组COL2A1、aggrecan mRNA和蛋白表达量均较对照组减少,且差异均有统计学意义(P均<0.05);TNF-α+Baf组COL2A1、aggrecan mRNA和蛋白表达量均较Baf组、TNF-α组减少,且差异均有统计学意义(P均<0.05);Baf组与TNF-α组COL2A1、aggrecan mRNA和蛋白表达量差异均无统计学意义(P均>0.05)。结论:自噬对大鼠腰椎间盘软骨终板细胞分泌的COL2A1和aggrecan具有保护作用。
Background:Intervertebral disc degeneration is considered as a major factor of low back pain.However,its underlying pathogenesis remains unclear.Autophagy may play an important role in intervertebral disc degeneration.Objective:To investigate the effect of autophagy on the expression of collagen typeⅡalpha 1(COL2A1)and aggrecan in rats’endplate chondrocytes.Methods:Endplate chondrocytes were treated with DMSO(control group),bafilomycin(Baf group),and TNF-α(TNF-αgroup)for 24 h,respectively.The autophagy in TNF-α+Baf group was inhibited by bafilomycin for 24 h before dealing with TNF-αin the next 24 h.The mRNA and protein expression of COL2A1 and aggrecan in rats’endplate chondrocytes were detected by real-time PCR,Western blotting analysis and immunofluorescence staining.Results:The mRNA and protein expression of COL2A1 and aggrecan in the Baf group,TNF-αgroup and TNF-α+Baf group were significantly lower than those in the control group(P<0.05).The mRNA and protein expression of COL2A1 and aggrecan in the TNF-α+Baf group were significantly lower than those in the Baf group and TNF-αgroup(P<0.05).There was no significant difference in the expression of COL2A1 or aggrecan between Baf group and TNF-αgroup(P>0.05).Conclusions:Autophgy can attenuate the inhibition of COL2A1 and aggrecan’s expression in endplate chondrocytes of rats.
作者
陈科
吕小华
林健静
孙军营
袁正彬
周辉
叶健文
李中杰
CHEN Ke;LYU Xiaohua;LIN Jianjing;SUN Junying;YUAN Zhengbin;ZHOU Hui;YE Jianwen;LI Zhongjie(Department of Spine Surgery,Shenzhen People's Hospital,The Second Clinical Medical College of Jinan University,The First Affiliated Hospital of Southern University of Science and Technology,Shenzhen 518020,Guangdong;College of Pharmacology,Guangdong Medical University,Zhanjiang 524023,Guangdong;Department of Orthopaedics,Peking University Shenzhen Hospital,Shenzhen 518036,Guangdong,China)
出处
《中华骨与关节外科杂志》
2021年第7期632-637,共6页
Chinese Journal of Bone and Joint Surgery
基金
广东省自然科学基金(2016A030310070)
深圳市人民医院中青年技术骨干科研培育项目(登峰计划)(SYKYPY201920)
