解剖学报 ›› 2024, Vol. 55 ›› Issue (5): 565-572.doi: 10.16098/j.issn.0529-1356.2024.05.007

• 细胞和分子生物学 • 上一篇    下一篇

小鼠冠状动脉粥样硬化区域炎性因子对干细胞迁移的调控作用

杨云1  孙攀文1  许亚平2,3  郭志坤1,2*
  

  1. 1.新乡医学院河南省医用组织再生重点实验室, 河南 新乡  453003; 2.郑州市第七人民医院河南省心脏重构与移植重点实验室,郑州  453000; 3.USM-ALPS Laboratory for Heart Research, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia
  • 收稿日期:2023-07-12 修回日期:2023-10-07 出版日期:2024-10-06 发布日期:2024-10-06
  • 通讯作者: 郭志坤 E-mail:gzk@xxmu.edu.cn
  • 基金资助:
    河南省自然科学基金;河南省医学科技攻关计划

Regulation of inflammatory factors in the coronary atherosclerosis region on stem cell migration in mice

YANG  Yun1  SUN  Pan-wen1  XU  Ya-ping2,3  GUO  Zhi-kun1,2*   

  1. 1.He’nan Key Laboratory of Medical Tissue Reneration, Xinxiang Medical University, He’nan Xinxiang  453003, China;  2.He’nan Key Laboratory of Cardiac Remodeling and Heart Transplantation, Zhengzhou Seventh People’s Hospital,Zhengzhou  45300,China; 3.USM-ALPS Laboratory for Heart Research, Advanced Medical and Dental Institute, University Sains Malaysia, Bertam, Penang, Malaysia
  • Received:2023-07-12 Revised:2023-10-07 Online:2024-10-06 Published:2024-10-06
  • Contact: GUO Zhi-kun E-mail:gzk@xxmu.edu.cn

摘要:

目的   探讨冠状动脉粥样硬化(AS)区域肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)炎性因子对干细胞抗原-1(Scal-1)和Nanog阳性细胞迁移的调控作用。 方法   取动脉粥样硬化模型小鼠和正常小鼠各10只,取出两组新鲜心脏,OCT包埋,冷冻切片,分别进行HE染色和油红O染色,观察冠状动脉病理变化;免疫组织化学染色,光学显微镜观察TNF-α、IL-6、Scal-1和Nanog在冠状动脉壁的表达变化。取20只出生1周小鼠的心室组织,利用密度梯度离心法提取原代Scal-1和Nanog阳性细胞,并纯化、培养,免疫荧光技术鉴定Scal-1和Nanog阳性细胞纯度。取第1代(P1)细胞进行Transwell迁移实验,观察不同浓度的炎症因子对Scal-1和Nanog阳性细胞迁移的影响。 结果   血脂检测证明,模型小鼠总胆固醇、甘油三脂、低密度脂蛋白胆固醇均明显高于正常对照组;组织学染色证明,所用小鼠模型均存在明显的冠状动脉粥样硬化。免疫组织化学结果显示,动脉粥样硬化区域炎症因子TNF-α和IL-6呈强阳性表达;同时,动脉粥样硬化斑块处Scal-1、Nanog阳性细胞较正常组增多。P1细胞中Scal-1的阳性率为80%,Nanog阳性率为91%。Transwell实验显示,低浓度IL-6对细胞迁移无作用,高浓度IL-6有抑制其迁移的作用;低浓度TNF-α促进细胞迁移,0.5μg/L TNF-α对细胞迁移影响最明显,在高浓度时起抑制迁移作用。 结论   TNF-α、IL-6和Scal-1、Nanog阳性细胞均参与AS的发生和发展,适当浓度的炎症因子可以增强干细胞向冠状动脉粥样硬化区域迁移。 

关键词: 干细胞抗原-1|Nanog|炎症因子|动脉粥样硬化|免疫组织化学|免疫荧光|细胞迁移实验|小鼠 

Abstract:

Objective   To investigate the effects of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in coronary arteriosclerosis (AS) on the migration of stem cell antigen-1(Scal-1) and Nanog positive cells migration.  Methods   Ten atherosclerotic model mice and 10 normal mice were taken respectively, and the fresh hearts of the two groups were taken out, OCT embedding, frozen sections, HE staining and oil red O staining were performed respectively to observe the pathological changes of coronary arteries; Using immunohistochemical staining techniques, the expression changes of TNF-α, IL-6, Scal-1, and Nanog in the coronary artery wall were observed under light microscope. The ventricular tissue of 20 1-week-old mice was excised. Primary Scal-1 and Nanog positive cells were extracted using density gradient centrifugation, and these cells were purified and cultured. Immunofluorescence technology was used to identify the purity of Scal-1 and Nanog positive cells. Transwell migration experiments on passage 1(P1) cells was performed to observe the effects of different concentrations of inflammatory factors on the migration of Scal-1 and Nanog positive cells.  Results  The blood lipid test showed that the total cholesterol, triglycerides, and low-density lipoprotein cholesterol of the model mice were significantly higher than those of the normal control group. Histological staining showed that all the mouse models had obvious coronary atherosclerosis plaque. Immunohistochemical staining results showed that the inflammatory factor TNF-α and IL-6 in atherosclerosis region were strong positive expression, while Scal-1 and Nanog positive cells in atherosclerotic plaques increased, compared to the normal group. The positive rate of Scal-1 in P1 generation cells was about 80%, The positive rate of Nanog was approximately 91%. The Transwell experiment showed that IL-6 had no effect on cell migration at low concentrations, but had an inhibitory effect on cell migration at high concentrations; Low concentrations TNF-α promoted cell migration with 0.5 μg/L TNF-α showing the most significant effection cell migration, and exhibiting inhibitcry effects inhibit at high concentrations.  Conclusion   TNF-α  and IL-6 inflammatory factors, Scal-1 and Nanog positive cells are all involved in the occurrence and development of AS, and the appropriate concentration of inflammatory factors can enhance the migration of stem cells to the coronary atherosclerosis region. 

Key words: Stem cell antigen-1|Nanog, Inflammatory factor|Atherosclerosis| Immunohistochemistry| Immunofluorescence|Cell migration experiment|Mouse

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