重组人白血病抑制因子蛋白对兔椎间盘退变模型髓核细胞凋亡的影响机制  

Effects of Recombinant Human Leukemia Inhibitory Factor Protein on Apoptosis of Nucleus Pulposus Cells in Rabbit Disc Degeneration Model

作　　者：穆林 房昕[1] 龚箭 高迎庆[1] 周强 MU Lin;FANG Xin;GONG Jian;GAO Yingqing;ZHOU Qiang(Dept of Orthopaedic Ward, Tieling Central Hospital, Liaoning Tieling 112000, China;Dept. of Orthopedics, the First Hospital Affiliated to Army Medical University, Chongqing 400013, China)

机构地区：[1]铁岭市中心医院骨科一病房,辽宁铁岭112000 [2]中国人民解放军陆军军医大学第一附属医院骨科,重庆400013

出　　处：《中国医院用药评价与分析》2020年第4期403-406,共4页Evaluation and Analysis of Drug-use in Hospitals of China

基　　金：国家自然科学基金(No.81772378)。

摘　　要：目的:探讨重组人白血病抑制因子(leukemia inhibitory factor,LIF)蛋白对兔椎间盘退变模型髓核细胞(nucleus pulposus cells,NPCs)凋亡的影响机制。方法:采用针刺法构建兔椎间盘退变模型,利用磁共振成像观察兔椎间盘退变程度,摘除髓核组织提取组织蛋白,采用Western Blot法检测髓核组织LIF蛋白表达水平,提取退变NPCs,采用流式细胞术观察不同浓度重组人LIF蛋白对退变NPCs凋亡的影响,采用Western Blot法检测不同浓度重组人LIF蛋白对细胞外基质成分和MAPK-ERK1/2通路蛋白表达的影响。结果:模型组兔的椎间盘高度和信号-噪声比(SNR)明显低于对照组,差异均有统计学意义(P<0.05);模型组兔髓核组织中LIF蛋白表达水平明显高于对照组,差异有统计学意义(P<0.05);针刺2周、4周及8周,各组兔髓核组织中LIF蛋白表达水平比较,差异有统计学意义(P<0.05);0、10、20、50及100 ng/ml重组人LIF蛋白处理的退变NPCs凋亡率比较,差异有统计学意义(P<0.05);0、10、20、50及100 ng/ml重组人LIF蛋白处理的退变NPCs中Aggtecan蛋白、Ⅱ型胶原及p-ERK1/2蛋白表达水平比较,差异有统计学意义(P<0.05)。结论:兔椎间盘退变髓核组织中LIF蛋白呈高表达,其可能通过激活ERK1/2通路促进细胞外基质合成达到抑制NPCs凋亡作用,重组LIF在延缓椎间盘退变方面显示出了良好前景。OBJECTIVE:To probe into the effects of recombinant human leukemia inhibitory factor(LIF)protein on apoptosis of nucleus pulposus cells in rabbit intervertebral disc degeneration model.METHODS:The rabbit intervertebral disc degeneration model was established by acupuncture.The degeneration degree of rabbit intervertebral disc was observed by magnetic resonance imaging.Tissue protein was extracted from nucleus pulposus tissue.The LIF protein expression level in nucleus pulposus tissue was detected by Western Blot.Degenerated nucleus pulposus cells(NPCs)were extracted.Flow cytometry was used to observe the effect of different concentrations of recombinant human LIF protein on apoptosis of degenerated NPCs.Western Blot was used to detect the effects of different concentrations of recombinant human LIF protein on extracellular matrix components and MAPK-ERK1/2 pathway protein expression.RESULTS:The intervertebral disc height and signal-to-noise ratio(SNR)in the model group were lower than those in the control group,with statistically significant difference(P<0.05).The expression level of LIF protein in nucleus pulposus tissue of model group was higher than that of control group,with statistically significant difference(P<0.05).The difference of LIF protein expression level in rabbit nucleus pulposus tissue after acupuncture for 2 weeks,4 weeks and 8 weeks was statistically significant(P<0.05).The apoptosis rate of degenerated NPCs treated with 0,10,20,50 and 100 ng/ml recombinant human LIF protein were compared,and the differences were statistically significant(P<0.05).0,10,20,50,and 100 ng/ml of restructuring LIF protein processing the degeneration of NPCs Aggtecan protein,collagen typeⅡand p-ERK1/2 protein expression level were compared,with statistically significant difference(P<0.05).CONCLUSIONS:LIF protein is highly expressed in the degenerated nucleus pulposus of rabbits,which may inhibit the apoptosis of nucleus pulposus cells by activating ERK1/2 pathway and promoting the synthesis of extracellular matrix.Re

关 键 词：重组人LIF蛋白 兔椎间盘退变模型 髓核细胞凋亡 机制 

分 类 号：R96[医药卫生—药理学]