[1]刘浠,易威威,温亚枫,等.锌指蛋白A20及其相关炎症因子在椎间盘髓核细胞退变前后的表达变化[J].第三军医大学学报,2016,38(18):2077-2081.
 Liu Xi,Yi Weiwei,Wen Yafeng,et al.Expression of zinc finger protein A20 and related inflammatory factors in nucleus pulposus cells in degenerative intervertebral disc[J].J Third Mil Med Univ,2016,38(18):2077-2081.
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锌指蛋白A20及其相关炎症因子在椎间盘髓核细胞退变前后的表达变化(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第18期
页码:
2077-2081
栏目:
临床医学
出版日期:
2016-09-30

文章信息/Info

Title:
Expression of zinc finger protein A20 and related inflammatory factors in nucleus pulposus cells in degenerative intervertebral disc
作者:
刘浠易威威温亚枫叶禾谭富强刘渤
重庆医科大学附属第一医院骨科
Author(s):
Liu XiYi WeiweiWen YafengYe HeTan FuqiangLiu Bo

Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing,400016, China

关键词:
椎间盘退变髓核细胞锌指蛋白A20炎症反应
Keywords:
disc degeneration nucleus pulposus zinc finger protein A20 inflammatory reaction
分类号:
R341;R363.22;R681.53
文献标志码:
A
摘要:

目的     检测椎间盘髓核细胞退变前后锌指蛋白A20及其相关炎症因子的表达变化。方法     获取我院骨科患者术中切除的椎间盘组织,分为正常组和退变组,采用阿辛蓝染色观察人椎间盘髓核细胞的退变情况,免疫组化观察人椎间盘髓核细胞A20、TNF-α、IL-1β及NF-κB p65蛋白的表达。体外培养胚胎小鼠髓核细胞,分为对照组(只加入等量的PBS)、内毒素(LPS)刺激组和NF-κB抑制组,分别以Real-time PCR检测LPS刺激细胞和加入NF-κB抑制剂后,A20、TNF-α、IL-1β、Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-4(ADAMST-4)、Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶-5(ADAMST-5)、基质金属蛋白酶-13(MMP-13)的表达变化;Western blot检测A20、TNF-α、IL-1β、ADAMTS-4、ADAMTS-5、MMP-13、p65、磷酸化p65(p-p65) 的表达变化。结果      退变组髓核细胞数量显著少于正常组,并有明显的聚集现象;退变组髓核细胞中A20、TNF-α、IL-1β、p65蛋白表达明显高于正常组;LPS刺激胚胎小鼠髓核细胞后,LPS刺激组A20、TNF-α、IL-1β、ADAMST-4、ADAMST-5、MMP-13 mRNA与蛋白的表达均高于NF-κB抑制组与对照组(P<0.05),A20、TNF-α、IL-1β、ADAMTS-4、ADAMTS-5、MMP-13、p65、p-p65蛋白含量也较NF-κB抑制组与对照组显著增高,差异有统计学意义(P<0.05)。结论     髓核细胞炎症反应与椎间盘退变的发生、发展密切相关, A20可能通过NF-κB信号通路对抑制髓核细胞退变时炎症反应起到关键作用。

Abstract:

Objective    To study the expression of zinc finger protein A20 and related inflammatory factors in nucleus pulposus cells in degenerative intervertebral disc. Methods      Human intervertebral disc tissues provided by department of orthopedics in our hospital were categorized into normal and degenerative groups. The degenerative degree of nucleus pulposus cells was evaluated by Alcian blue staining. The expression of A20, TNF-α, IL-1β and p65 in nucleus pulposus cells was detected by immunohistochemical (IHC) staining. In vitro embryonic mouse nucleus pulposus cells were cultured. Based on the treatment, the cells were divided into control group, LPS stimulation group and NF-κB inhibition group. Real-time PCR (RT-PCR) was performed to measure the mRNA expression of A20, TNF-α, IL-1β, ADAMST-4, ADAMST-5, and MMP-13. Western blotting was used to detect the protein expression of A20, TNF-α, IL-1β, ADAMTS-4, ADAMTS-5, MMP-13, p65, and p-p65. Results     In human degenerative intervertebral disc, the number of nucleus pulposus cells was obviously decreased in favor of aggregation as compared to the normal group. Also, the expression of A20 was significantly higher than the normal group detected by IHC staining. In embryonic mouse nucleus pulposus cells, the mRNA expression of A20, TNF-α, IL-1β, ADAMST-4, ADAMST-5 and MMP-13 was significantly higher in LPS stimulation group than that in NF-kB inhibition group and control group (P<0.05). Likely, LPS stimulation increased the protein levels of A20, TNF-α, IL-1β, ADAMST-4, ADAMST-5, MMP-13, p65 and p-p65 compared to NF-kB inhibition group and control group, which was of statistic significance (P<0.05). Conclusion     The inflammatory response of nucleus pulposus cells is closely related to the occurrence and development of intervertebral disc degeneration, which A20 may play an important role in inhibiting this inflammatory reaction through NF-kB signaling pathway.

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更新日期/Last Update: 2016-09-21