脉冲电磁场对椎间盘退行性病变大鼠髓核神经肽Y、细胞凋亡及基质降解的影响  

The effects of pulsed electromagnetic irradiation on neuropeptide Y regulation,the apoptosis of nucleus pulposus cells and degradation of the extracellular matrix in rats with intervertebral disc degeneration

作　　者：王正坤 姚智 魏梦诚 张诗爽 周俊龙 黎清波 蔡磊 周传坤 寇博文 刘伟军 Wang Zhengkun;Yao Zhi;Wei Mengcheng;Zhang Shishuang;Zhou Junlong;Li Qingbo;Cai Lei;Zhou Chuankun;Kou Buen;Liu Weijun(Deprtment of Spine Surgery,Wuhan Fourth Hospital,Wuhan 430030,China;Department of Spine Surgery,Puai Hospital,Tongji Medical College Affiliated to Huazhong Unirersiy of Scierece and Technology,Wuhan 430030,China;Schoot of Medicine,Jianghan Unirersity,Wuhan 430056,China)

机构地区：[1]武汉市第四医院脊柱及骨肿瘤外科中心,武汉430030 [2]华中科技大学同济医学院附属武汉普爱医院脊柱及骨肿瘤外科中心,武汉430030 [3]江汉大学医学院,武汉430056

出　　处：《中华物理医学与康复杂志》2024年第7期601-607,共7页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：湖北省自然科学基金项目(2021CFB522);武汉市科技局知识创新专项-曙光计划项目(2023020201020550)。

摘　　要：目的观察脉冲电磁场(PEMF)对椎间盘退行性病变(IDD)大鼠髓核组织及离体髓核细胞神经肽Y(NPY)的调控作用,并探讨其对髓核细胞凋亡及基质降解的影响。方法采用随机数字表法将18只SD大鼠分为对照组、IDD模型组(简称模型组)和PEMF组。将模型组及PEMF组大鼠制成IDD动物模型,PEMF组于造模后给予PEMF干预,每天曝磁1次,连续干预14 d。同期体外培养大鼠原代髓核细胞并分为对照组、TNF-α模型组(简称模型组)和TNF-α+PEMF组(简称PEMF组),分别向模型组、PEMF组细胞培养板内注入25μL TNF-α(终浓度为50 ng/ml),PEMF组随后给予曝磁处理,每天曝磁1次,连续干预6 d,对照组则同期向细胞培养板内注入25μL磷酸盐缓冲液(PBS)。于造模8周后采用番红固绿染色评估各组大鼠椎间盘组织病理形态学改变;同时检测各组大鼠椎间盘及离体髓核细胞NPY、神经肽Y2受体(NPY2R)、B细胞淋巴瘤/白血病-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2因子(Bcl-2)、Ⅱ型胶原(Col-Ⅱ)、基质金属蛋白酶-3(MMP3)的表达水平。结果模型组椎间盘纤维组织出现断裂、褶皱,且多糖、蛋白质等成分明显减少,骨纤维增多。PEMF组椎间盘纤维组织断裂较少,软骨组织增多,纤维环与髓核边界较清晰。模型组大鼠椎间盘纤维环及髓核组织中均有NPY表达,且NPY、NPY2R表达量均较对照组明显增加(P<0.05),PEMF组NPY、NPY2R表达量较模型组进一步升高(P<0.05)。与对照组比较,模型组Bax含量显著增加(P<0.05),Bcl-2表达明显减少(P<0.05),而PEMF组Bax含量较模型组明显降低(P<0.05)。模型组Col-Ⅱ水平显著低于对照组(P<0.05),MMP3蛋白表达则明显增强(P<0.05);PEMF组Col-ⅡmRNA表达较模型组显著升高(P<0.05),MMP3蛋白表达明显降低(P<0.05)。各组离体髓核细胞上述指标检测结果与在体实验观察结果基本一致。结论PEMF干预可能通过上调NPY表达,阻滞Bax/Bcl-2信号通路抑制髓核细胞凋亡Objective To document any effect of a pulsed electromagnetic field(PEMF)on the regulation of neuropeptide Y(NPY)in nucleus pulposus(NP)tissue,NP cell apoptosis and matrix degradation using rats with intervertebral disc degeneration(IDD).Methods Eighteen Sprague-Dawley rats were randomly divided into a control group,an IDD model group(the model group),and a PEMF group.IDD was induced in both the model and PEMF groups.Right after the modeling,the PEMF group received 14 days of PEMF treatment,while the control group and model group were given no special treatment.Meanwhile,the primary rat nucleus pulposus cells(NPCs)were cultured using Dulbecco′s Modified Eagle Medium at 37℃and 5%CO2.When the fusion rate reached 90%after passage,the NPCs were divided into a control group,a TNF-αmodel group(referred to as model group)and TNF-α+PEMF group(referred to as PEMF group)and treated accordingly.Eight weeks after the modeling,safranin-o/fast green staining was used to assess any pathological morphology changes.The expression of NPY,neuropeptide Y receptor Y2(NPY2R),bcl-2-associated X protein(Bax),B-cell lymphoma 2(Bcl-2),collagen type II(Col-II)and matrix metalloproteinase-3(MMP3)in the intervertebral disc and the cultivated nucleus pulposus cells of the 3 groups were determined.Results The intervertebral disc cells in the model group were ruptured and folded,with significantly increased polysaccharide and protein components,and significantly increased bone fibers.In the PEMF group the cell boundaries were clearer,with less fibrin fracture and increased cartilage tissue.NPY was expressed in the fibrous annulus and the nucleus pulposus of the intervertebral disc in the model group.The average expression levels of NPY and NPY2R were significantly higher than in the control group and the model group.Compared with the control group,there was a significant increase in the level of Bax and a significant decrease in the expression of Bcl-2 in the model group,and there was a significant decrease in the level of Bax in the PEMF

关 键 词：脉冲电磁场 神经肽Y 椎间盘退变 凋亡 

分 类 号：R681.53[医药卫生—骨科学]