中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (28): 4469-4474.doi: 10.3969/j.issn.2095-4344.2017.28.007

• 脊柱组织构建 spinal tissue construction • 上一篇    下一篇

丝裂原活化蛋白激酶信号通路在骨桥蛋白介导黄韧带骨化中的作用

李学斌,许 政,周盛源,刘晓东,王智清,许国峰,陈雄生   

  1. 解放军第二军医大学附属上海长征医院脊柱三科,上海市 200003
  • 修回日期:2017-06-20 出版日期:2017-10-08 发布日期:2017-11-10
  • 通讯作者: 陈雄生,博士,主任医师,解放军第二军医大学附属长征医院脊柱三科,上海市 200003
  • 作者简介:李学斌,男,1989年生,山东省烟台市人,汉族, 2016年解放军第二军医大学毕业,硕士,医师。
  • 基金资助:

    国家自然科学基金面上项目(81171753);上海市科学技术委员会项目(15140903800)

Role of MAPK signaling pathway in osteopontin-mediated ossification of the ligamentum flavum

Li Xue-bin, Xu Zheng, Zhou Sheng-yuan, Liu Xiao-dong, Wang Zhi-qing, Xu Guo-feng, Chen Xiong-sheng   

  1. Third Department of Spinal Surgery, Shanghai Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China
  • Revised:2017-06-20 Online:2017-10-08 Published:2017-11-10
  • Contact: Chen Xiong-sheng, M.D., Chief physician, Third Department of Spinal Surgery, Shanghai Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China
  • About author:Li Xue-bin, Master, Physician, Third Department of Spinal Surgery, Shanghai Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81171753; the Project of Science and Technology Commission of Shanghai, No. 15140903800

摘要:

文章快速阅读:

文题释义:
黄韧带骨化:
黄韧带骨化是发生在脊柱的异位骨化,近年来作为一种独立的临床疾病被国内外学者广泛重视,多发于东南亚,好发年龄50-60岁。其好发于下胸椎及胸腰段脊柱,可致椎管严重狭窄引起脊髓压迫损伤,常导致脊髓严重的不可逆性伤害,目前手术是惟一的解决方法。其发病机制尚不明确,目前研究表明脊柱局部机械应力、激素以及肥胖与其发病有关。
丝裂原活化蛋白激酶信号通路:丝裂原活化蛋白激酶信号通路是生物体内重要的信号转导系统之一,它可以被物理应激、炎性细胞因子、生长因子以及细胞复合物等一系列细胞外信号或刺激激活,从而参与介导细胞生长、发育、分裂以及分化等多种生理及病理过程。在哺乳动物细胞中丝裂原活化蛋白激酶信号的主要信号分子包括:ERK1/2,JNK及p38等。

 

摘要
背景:
丝裂原活化蛋白激酶(MAPK)信号通路是生物体内重要的信号转导系统之一。课题组前期动物实验提示骨桥蛋白可介导黄韧带骨化的发生。
目的:观察MAPK信号通路在骨桥蛋白介导黄韧带骨化中的作用。
方法:将16例患者后路椎板切除手术中取得黄韧带标本分为黄韧带骨化以及非骨化两组,每组8例。通过免疫组织化学染色观察骨桥蛋白及其受体CD44、整合素的表达;利用骨桥蛋白对黄韧带细胞进行诱导,并利用蛋白印记方法检测丝裂素活化蛋白激酶信号通路分子的磷酸化激活情况,随后再利用SB203580和U0126阻滞剂分别阻断丝裂素活化蛋白激酶相应信号通路观察骨桥蛋白的诱导情况。
结果与结论:①骨桥蛋白及其受体CD44及整合素的表达:骨化组的标本中均检测到骨桥蛋白及其受体CD44的表达,但未检测到整合素的表达,非骨化组中则未见到骨桥蛋白及其受体的表达;②MAPK信号通路变化:在骨桥蛋白的诱导作用下黄韧带细胞中骨化指标碱性磷酸酶及骨钙素表达增加(P < 0.05),同时骨桥蛋白能激活丝裂素活化蛋白激酶信号通路中的p38及ERK1/2信号分子的磷酸化(P < 0.05),但JNK氨基末端激酶的磷酸化激活并不明显。利用SB203580阻滞剂阻断p38的磷酸化可明显阻断骨桥蛋白诱导的黄韧带细胞的骨向分化(P < 0.05),而U0126阻滞剂阻断效果不明显;③结果证实,MAPK信号通路在骨桥蛋白介导黄韧带骨化中其重要作用,p38是诱导黄韧带细胞骨向分化的关键因子。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID:
0000-0002-9487-3886(陈雄生)

关键词: 组织构建, 组织工程, 黄韧带骨化症, 骨桥蛋白, 丝裂素活化蛋白激酶信号通路, 黄韧带细胞, 国家自然科学基金

Abstract:

BACKGROUND: Mitogen-activated protein kinase (MAPK) signaling pathway is an important signal transduction system. Our precious animal experiments have shown that osteopontin can mediate the ossification of the ligamentum flavum.
OBJECTIVE: To investigate the role of MAPK signaling pathway in osteopontin-induced ossification of the ligamentum flavum.
METHODS: The ligamentum flavum specimens obtained from 16 cases undergoing thoracic/lumbar posterior decompression surgery were divided into ossification and non-ossification groups (n=8 per group). The expression of osteopontin and its receptors CD44 and integrin was observed by immunohistochemical staining. The activation of phosphorylation in MAPK signaling pathway was detected by western blot assay. The MAPK signaling pathway was blocked by SB203580 or U0126 blocker alone to observe the induction of osteopontin.
RESULTS AND CONCLUSION: Osteopontin and its receptor CD44 were expressed in the ossification group, but not in the non-ossififcation group. However, the expression of integrin was not detected in the ossification group. The expression levels of alkaline phosphatase and osteocalcin in the ligamentum flavum were significnatly increased under the induction of osteopontin (P < 0.05), and osteopontin could activate the phosphorylation of P38 and ERK1/2 in the MAPK signaling pathway (P < 0.05), but the phosphorylation of JNK was not obvious. p38 phosphorylation blocked with SB203580 blocker could significantly inhibit the osteopontin-induced osteoblast differentiation of ligament flavum cells (P < 0.05), while U0126 blocker had no obvious effect. These results indicate that p38 in MAPK signaling pathway is a key molecule in osteopontin-mediated ossification of the ligamentum flavum.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Tissue Engineering, Ligmentum Flavum, Alkaline Phosphatase

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