中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (7): 1624-1631.doi: 10.12307/2026.589

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

血红素氧合酶1减轻脂多糖诱导髓核间充质干细胞的炎症反应

蔡子鸣1，于庆贺2，马鹏飞2，张  鑫2，周龙千1，张崇阳1，林文平2   

  1. 1广州中医药大学附属深圳平乐骨伤科医院，广东省深圳市  518118；2广州中医药大学附属深圳平乐骨伤科医院脊柱外科，广东省深圳市  518118
  • 收稿日期:2024-12-18 修回日期:2025-05-09 接受日期:2025-05-29 出版日期:2026-03-08 发布日期:2025-08-18
  • 通讯作者: 林文平，博士，主任医师，博士生导师，广州中医药大学附属深圳平乐骨伤科医院脊柱外科，广东省深圳市 518118
  • 作者简介:蔡子鸣，男，2000年生，江西省南昌市人，汉族，广州中医药大学在读硕士，主要从事中西医结合治疗脊柱疾病的研究。 并列第一作者：于庆贺，男，1992年生，河北省邢台市人，汉族，2020年南方医科大学毕业，硕士，医师，主要从事脊柱脊髓损伤修复方面的研究。
  • 基金资助:
    国家自然科学基金面上项目(81771323)，项目负责人：林文平；广东省自然科学基金面上项目(2021A1515010722，2024A1515010445)，项目负责人：林文平；深圳市自然科学基金面上项目(JCYJ20190813112401660)，项目负责人：林文平

Heme oxygenase-1 alleviates lipopolysaccharide-induced inflammatory response in nucleus pulposus mesenchymal stem cells

Cai Ziming1, Yu Qinghe2, Ma Pengfei2, Zhang Xin2, Zhou Longqian1, Zhang Chongyang1, Lin Wenping2   

  1. 1Shenzhen Pingle Orthopedic Hospital Affiliated to Guangzhou University of Chinese Medicine, Shenzhen 518118, Guangdong Province, China; 2Department of Spine Surgery, Shenzhen Pingle Orthopedic Hospital Affiliated to Guangzhou University of Chinese Medicine, Shenzhen 518118, Guangdong Province, China
  • Received:2024-12-18 Revised:2025-05-09 Accepted:2025-05-29 Online:2026-03-08 Published:2025-08-18
  • Contact: Lin Wenping, MD, Chief physician, Doctoral supervisor, Department of Spine Surgery, Shenzhen Pingle Orthopedic Hospital Affiliated to Guangzhou University of Chinese Medicine, Shenzhen 518118, Guangdong Province, China
  • About author:Cai Ziming, Master candidate, Shenzhen Pingle Orthopedic Hospital Affiliated to Guangzhou University of Chinese Medicine, Shenzhen 518118, Guangdong Province, China Yu Qinghe, MS, Physician, Department of Spine Surgery, Shenzhen Pingle Orthopedic Hospital Affiliated to Guangzhou University of Chinese Medicine, Shenzhen 518118, Guangdong Province, China Cai Ziming and Yu Qinghe contributed equally to this article.
  • Supported by:
    National Natural Science Foundation of China, No. 81771323 (to LWP); Natural Science Foundation of Guangdong Province of China, No. 2021A1515010722 and No. 2024A1515010445 (to LWP); Natural Science Foundation of Shenzhen Municipality, No. JCYJ20190813112401660 (to LWP)


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摘要：

文题释义：

血红素氧合酶1：是血红素代谢的关键酶，广泛存在于各类组织中，能够促进血红素分解，产生一氧化碳、铁离子和胆绿素，还具有显著的抗炎效果及强大的抗氧化特性。
髓核间充质干细胞：具备多向分化能力和自我更新的特性，是治疗椎间盘退变最具潜力的细胞类型。

摘要
背景：研究表明，血红素氧合酶1具有抗炎、抗凋亡作用，但它能否在髓核间充质干细胞中发挥抗炎保护作用尚不明确。
目的：探讨血红素氧合酶1对炎症微环境下髓核间充质干细胞的保护作用及机制。
方法：①从SD大鼠尾椎椎间盘中提取原代髓核间充质干细胞，通过流式细胞术、三系分化进行鉴定。②采用血红素氧合酶1过表达慢病毒感染髓核间充质干细胞，通过荧光显微镜观察绿色荧光蛋白表达以及Western blot检测血红素氧合酶1蛋白表达水平评估感染效率。③髓核间充质干细胞分为4组：对照组用DMEM/F-12完全培养基培养24 h，模型组、空载体组、血红素氧合酶1过表达组分别为未感染细胞、感染LV-Ctrl细胞、感染LV-HO-1细胞，3组均用添加5 μg/mL脂多糖的DMEM/F-12完全培养基培养24 h。采用Western blot、免疫荧光检测炎症相关蛋白表达，Western blot检测核因子κB信号通路蛋白水平。

结果与结论：①大鼠髓核间充质干细胞排列紧密，呈贴壁生长，多为纺锤形或梭形，通过流式细胞术检测髓核间充质干细胞具有较高的纯度，三系分化鉴定髓核间充质干细胞具有向脂肪细胞、骨细胞、软骨细胞分化的能力；②血红素氧合酶1过表达慢病毒感染的髓核间充质干细胞表达绿色荧光蛋白，血红素氧合酶1蛋白表达显著升高；③与模型组相比，血红素氧合酶1过表达组NOD样受体热蛋白结构域相关蛋白3、基质金属蛋白酶13、基质金属蛋白酶3、肿瘤坏死因子α蛋白表达均明显降低(P < 0.05)；④与模型组相比，血红素氧合酶1过表达组磷酸化核因子κB/核因子κB比值显著下降(P < 0.05)，磷酸化核因子κB抑制蛋白/核因子κB抑制蛋白也显著下降(P < 0.05)。以上结果表明，血红素氧合酶1通过抑制核因子κB信号通路的激活，有效减少了脂多糖诱导的髓核间充质干细胞炎症反应。

https://orcid.org/0000-0001-5029-5466(林文平)

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: 血红素氧合酶1,  髓核间充质干细胞,  脂多糖,  炎症,  椎间盘退变,  核因子κB

Abstract: BACKGROUND: Studies have shown that heme oxygenase-1 has anti-inflammatory and anti-apoptotic effects. However, its potential to exert anti-inflammatory protective effects in nucleus pulposus mesenchymal stem cells remains unclear.
OBJECTIVE: To explore the protective effects and mechanisms of heme oxygenase-1 on nucleus pulposus mesenchymal stem cells under an inflammatory microenvironment. 
METHODS: (1) Primary nucleus pulposus mesenchymal stem cells were isolated from the intervertebral disc of SD rat tails and identified by flow cytometry and trilineage differentiation. (2) Nucleus pulposus mesenchymal stem cells were infected with heme oxygenase-1 overexpression lentivirus, and green fluorescent protein expression was observed under a fluorescence microscope. Western blot assay was used to assess the heme oxygenase-1 protein expression levels and infection efficiency. (3) Nucleus pulposus mesenchymal stem cells were divided into four groups: the control group cells were cultured with DMEM/F-12 complete medium for 24 hours. The model group, empty vector group, and heme oxygenase 1 overexpression group were uninfected cells, LV-Ctrl cells infected, and LV-HO-1 cells infected, respectively, and cultured with DMEM/F-12 complete medium supplemented with 5 μg/mL lipopolysaccharide for 24 hours. Western blot assay and immunofluorescence were used to detect the expression of inflammation-related proteins. The expression levels of proteins related to the nuclear factor κB signaling pathway were assessed using western blot assay.  
RESULTS AND CONCLUSION: (1) Rat nucleus pulposus mesenchymal stem cells exhibited adherent growth, dense arrangement, vigorous proliferation, and spindle-shaped morphology. Flow cytometry results showed high purity of the cultured nucleus pulposus mesenchymal stem cells, and trilineage differentiation assay indicated the nucleus pulposus mesenchymal stem cells had good potential to differentiate into adipocytes, osteocytes, and chondrocytes. (2) The nucleus pulposus mesenchymal stem cells infected with heme oxygenase 1 overexpression lentivirus expressed green fluorescent protein, and heme oxygenase 1 was highly expressed. (3) Compared with the model group, the expression levels of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), matrix metalloproteinase 13, matrix metalloproteinase 3, and tumor necrosis factor alpha were significantly reduced in the heme oxygenase-1 overexpression group (P < 0.05). (4) Compared with the model group, the ratio of phosphorylated nuclear factor κB/nuclear factor κB was significantly decreased (P < 0.05), and phosphorylated nuclear factor κB inhibitor protein/nuclear factor κB inhibitor protein also decreased significantly (P < 0.05) in the heme oxygenase-1 overexpression group. The above results indicate that heme oxygenase 1 effectively reduces the lipopolysaccharide-induced inflammatory response of nucleus pulposus mesenchymal stem cells by inhibiting the activation of the nuclear factor κB signaling pathway. 

Key words: heme oxygenase-1,  , nucleus pulposus mesenchymal stem cell,  , lipopolysaccharide,  , inflammation,  , intervertebral disc degeneration,  , nuclear factor κB

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