中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (31): 4967-4974.doi: 10.12307/2022.993

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

人羊膜上皮细胞激活EGFR/ERK1信号轴调节关节软骨细胞增殖、凋亡和细胞外基质的合成

王  康1，2，智晓东1，2，张玉强1，2，龚  超1，2，王晨亮1，2，王  伟1，2，3   

  1. 1锦州医科大学附属第一医院骨科，辽宁省锦州市  121000；2辽宁省医学组织工程重点实验室，辽宁省锦州市  121000；3锦州医科大学骨外科学研究所，辽宁省锦州市  121000
  • 收稿日期:2021-07-05 接受日期:2021-08-19 出版日期:2022-11-08 发布日期:2022-04-24
  • 通讯作者: 王伟，博士，主任医师，教授，锦州医科大学附属第一医院骨科，辽宁省锦州市 121000；锦州医科大学骨外科学研究所，辽宁省锦州市 121000；辽宁省医学组织工程重点实验室，辽宁省锦州市 121000
  • 作者简介:王康，男，1995 年生，安徽省潜山县人，汉族，锦州医科大学在读硕士，主要从事软骨损伤修复相关研究。
  • 基金资助:
    锦州医科大学校企合作基金项目(2020002)，项目负责人：王伟；辽宁省科学技术基金项目 (20180551216)，项目负责人：智晓东

Human amniotic epithelial cells regulate the proliferation, apoptosis and extracellular matrix synthesis of articular chondrocytes by activating the EGFR/ERK1 signaling axis

Wang Kang1, 2, Zhi Xiaodong1, 2, Zhang Yuqiang1, 2, Gong Chao1, 2, Wang Chenliang1, 2, Wang Wei1, 2, 3   

  1. 1Department of Orthopedics, First Affiliated Hospital of Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China; 2Liaoning Key Laboratory of Medical Tissue Engineering, Jinzhou 121000, Liaoning Province, China; 3Institute of Orthopedics, Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China
  • Received:2021-07-05 Accepted:2021-08-19 Online:2022-11-08 Published:2022-04-24
  • Contact: Wang Wei, MD, Chief physician, Professor, Department of Orthopedics, First Affiliated Hospital of Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China; Liaoning Key Laboratory of Medical Tissue Engineering, Jinzhou 121000, Liaoning Province, China; Institute of Orthopedics, Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China
  • About author:Wang Kang, Master candidate, Department of Orthopedics, First Affiliated Hospital of Jinzhou Medical University, Jinzhou 121000, Liaoning Province, China; Liaoning Key Laboratory of Medical Tissue Engineering, Jinzhou 121000, Liaoning Province, China
  • Supported by:
    The School-Enterprise Cooperation Fund Project of Jinzhou Medical University, No. 2020002 (to WW); Science and Technology Foundation of Liaoning Provincial Department of Science and Technology, No. 20180551216 (to ZXD)

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摘要：

文题释义：
人羊膜上皮细胞：是近年来在干细胞治疗领域广受关注的“种子细胞”之一，由受精卵发育早期的囊胚内细胞团分化而来，兼具原始的胚胎干细胞特性和上皮细胞样形态结构，在不同外源性因子作用下，具有向神经细胞、骨细胞、心肌细胞、脂肪细胞等三胚层细胞分化的潜能。
细胞治疗在骨性关节炎中的应用价值：骨关节炎是一种晚期关节软骨损伤，其临床表现包括关节疼痛、关节畸形和关节活动受限，将干细胞直接注入关节腔或将含干细胞的材料植入体内可有效治疗轻度至中度软骨损伤，促进透明软骨的再生，改善患者生活质量。

背景：以往研究发现，人羊膜上皮细胞具有促进骨关节炎修复的作用，但人羊膜上皮细胞对软骨再生的作用机制尚未明确。
目的：探讨人羊膜上皮细胞对软骨细胞增殖、凋亡及细胞外基质合成能力的影响及其机制。
方法：酶消化法分离人羊膜上皮细胞。将人羊膜上皮细胞、关节软骨细胞以及表皮生长因子受体信号通路抑制剂AG1478在Transwell小室中共培养，通过CCK-8、流式、RT-qPCR检测软骨细胞增殖、凋亡和细胞外基质合成能力，Western blot检测表皮生长因子受体信号通路相关蛋白的表达。
结果与结论：①与人羊膜上皮细胞共培养后，软骨细胞的增殖活性明显增强，凋亡率明显降低， COL2，COL1，Aggrecan和SOX-9 mRNA表达水平显著增加，p-ERK1的蛋白表达水平显著提高；②加入表皮生长因子受体信号通路抑制剂AG1478后，软骨细胞的增殖活性明显减弱，凋亡率显著增加，COL2，COL1，Aggrecan和SOX-9的mRNA表达水平下降，p-ERK1的蛋白表达水平明显降低；③结果表明，人羊膜上皮细胞可以促进软骨细胞的增殖和细胞外基质的合成，并抑制其凋亡，其作用可能是通过EGFR/ERK1信号轴进行调节的。
缩略语：表皮生长因子受体：epidermal growth factor receptor，EGFR；细胞外信号调节激酶1：extracellular regulated protein kinases1，ERK1

https://orcid.org/0000-0003-2572-5975 (王伟) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 骨关节炎, 人羊膜上皮细胞, 软骨细胞, 增殖, 凋亡, 细胞外基质

Abstract: BACKGROUND: Previous studies have found that human amniotic epithelial cells have the effect of promoting the repair of osteoarthritis, but the mechanism of action of human amniotic epithelial cells on cartilage regeneration is not yet clear.  
OBJECTIVE: To explore the effect of human amniotic epithelial cells on chondrocyte proliferation, apoptosis and extracellular matrix synthesis and its mechanism. 
METHODS: The human amniotic epithelial cells were separated by enzymatic digestion. Human amniotic epithelial cells, articular chondrocytes, and epidermal growth factor receptor signaling pathway inhibitor (AG1478) were co-cultured in a Transwell chamber. The chondrocyte proliferation, apoptosis, and extracellular matrix synthesis were detected using CCK-8, flow cytometry, and RT-qPCR, respectively. The expression of related proteins in the epidermal growth factor receptor signaling pathway was determined by western blot assay.  
RESULTS AND CONCLUSION: (1) After co-culture with human amniotic epithelial cells, the proliferation of chondrocytes was significantly enhanced; the apoptotic rate was significantly reduced; the expression levels of COL2, COL1, Aggrecan, and SOX-9 mRNA were significantly increased; and the protein expression level of p-ERK1 was significantly increased. (2) After the addition of epidermal growth factor receptor signaling pathway inhibitor AG1478, the proliferation of chondrocytes was significantly weakened; the apoptotic rate was significantly increased; COL2, COL1, Aggrecan, and SOX-9 mRNA expression decreased; protein expression level of p-ERK1 was significantly reduced. (3) It is concluded that human amniotic epithelial cells can promote the proliferation of chondrocytes and the synthesis of extracellular matrix, and inhibit their apoptosis, and its effect may be regulated by the EGFR/ERK1 signaling axis. 

Key words: osteoarthritis, human amniotic epithelial cells, chondrocytes, proliferation, apoptosis, extracellular matrix

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