中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (31): 6625-6633.doi: 10.12307/2025.556

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

人脐带间充质干细胞共培养联合人参皂苷Rg1对心力衰竭细胞模型的影响

任疏桐1，郝  苗2，刘  越3，侯  平1，4，全娟花5   

  1. 1辽宁中医药大学，辽宁省沈阳市   110000；2沈阳市第四人民医院中医科，辽宁省沈阳市   110031；3沈阳医学院中医教研室，辽宁省沈阳市   110031；4辽宁中医药大学附属医院，辽宁省沈阳市   110000；5广东医科大学附属医院， 广东省湛江市   524000
  • 收稿日期:2024-07-18 接受日期:2024-09-26 出版日期:2025-11-08 发布日期:2025-02-18
  • 通讯作者: 侯平，硕士研究生，教授，博士生导师，辽宁中医药大学，辽宁省沈阳市 110000；辽宁中医药大学附属医院，辽宁省沈阳市 110000
  • 作者简介:任疏桐，女，1990年生，辽宁中医药大学在读博士，主治医师，主要从事中西医结合心血管方向研究。
  • 基金资助:
    国家自然科学基金项目(81971389)，项目负责人：全娟花

Effect of human umbilical cord mesenchymal stem cells co-culture combined with ginsenoside Rg1 on heart failure cell model

Ren Shutong1, Hao Miao2, Liu Yue3, Hou Ping1, 4, Quan Juanhua5   

  1. 1Liaoning University of Traditional Chinese Medicine, Shenyang 110000, Liaoning Province, China; 2Department of Traditional Chinese Medicine, Shenyang Fourth People’s Hospital, Shenyang 110031, Liaoning Province, China; 3Traditional Chinese Medicine Teaching and Research Office of Shenyang Medical University, Shenyang 110031, Liaoning Province, China; 4Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110000, Liaoning Province, China; 5Affiliated Hospital of Guangdong Medical University, Zhanjiang 524000, Guangdong Province, China
  • Received:2024-07-18 Accepted:2024-09-26 Online:2025-11-08 Published:2025-02-18
  • Contact: Hou Ping, MS, Professor, Doctoral supervisor, Liaoning University of Traditional Chinese Medicine, Shenyang 110000, Liaoning Province, China; Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110000, Liaoning Province, China
  • About author:Ren Shutong, Doctoral candidate, Attending physician, Liaoning University of Traditional Chinese Medicine, Shenyang 110000, Liaoning Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81971389 (to QJH)

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摘要：

文题释义：

人参皂苷Rg1：是人参中分离出来的皂苷，也是人参的主要生物活性成分。人参皂苷Rg1具有抗氧化、抗炎、抗血管生成和抗凋亡等活性。另外，人参皂苷Rg1可预防多种心血管疾病，已被报道具有抗心衰作用，例如人参皂苷Rg1通过线粒体自噬改善心力衰竭患者心脏重构。
人脐带间充质干细胞：指存在于新生儿脐带组织中的一种多功能干细胞，具有来源丰富、无痛采集、自我更新快、多向分化潜能、合成和分泌一系列营养因子和细胞因子、支持其他细胞的扩增、向病理区域迁移和归巢以及易于通过常规方法进行转染等优点，在再生领域备受关注。目前人脐带间充质干细胞移植需要解决的主要问题是促进细胞扩增、减少细胞损失、提高细胞归巢率、精确调节分化以及减少细胞衰老和凋亡。

摘要
背景：目前，如何促进细胞扩增、减少细胞损失、提高细胞归巢率、减少细胞凋亡是人脐带间充质干细胞临床前研究的主要问题。人参皂苷Rg1可在体外或体内促进间充质干细胞在不同微环境中的增殖、分化，其可能是提高人脐带间充质干细胞移植效率的候选药物。
目的：探讨人脐带间充质干细胞共培养联合人参皂苷Rg1对戊巴比妥钠诱导的心衰细胞模型的影响。
方法：将H9C2细胞分为5组：对照组、戊巴比妥钠组、人脐带间充质干细胞组、人参皂苷Rg1组、人脐带间充质干细胞+人参皂苷Rg1组。对照组H9C2细胞用正常DMEM培养基培养24 h，其他组H9C2细胞用含0.8%戊巴比妥钠的DMEM培养基培养7 min构建心衰细胞模型。建模后，分别用人脐带间充质干细胞、人参皂苷Rg1单独或共同处理。采用CCK-8法和EdU染色检测细胞增殖情况，TUNEL法检测细胞凋亡情况，按试剂盒说明检测Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性，ELISA测定上清液中肿瘤坏死因子α、白细胞介素1β和白细胞介素6水平，RT-qPCR测定细胞中肿瘤坏死因子α、白细胞介素1β、白细胞介素6、Bax和Bcl-2 mRNA表达，Western blot测定细胞中Bax、Bcl-2、Toll样受体4、p65和p-p65蛋白表达。
结果与结论：①与戊巴比妥钠组比较，人脐带间充质干细胞组、人参皂苷Rg1组和人脐带间充质干细胞+人参皂苷Rg1组H9C2细胞活力和EdU阳性率升高，TUNEL阳性率以及Bax mRNA和蛋白表达降低，Bcl-2 mRNA和蛋白表达升高，Na+-K+-ATP酶活性降低，Ca2+-Mg2+-ATP酶活性升高，H9C2细胞上清液中肿瘤坏死因子α、白细胞介素1β和白细胞介素6水平降低，H9C2细胞中肿瘤坏死因子α、白细胞介素1β和白细胞介素6 mRNA表达降低，Toll样受体4和p-p65蛋白表达降低，差异均有显著性意义(P < 0.05)；②与人脐带间充质干细胞组和人参皂苷Rg1组比较，人脐带间充质干细胞+人参皂苷Rg1组上述指标进一步改善(P < 0.05)。结果表明：人脐带间充质干细胞联合人参皂苷Rg1提高了戊巴比妥钠诱导的心衰细胞活力，并抑制了Toll样受体4/核因子κB通路介导的炎症。

关键词: 人脐带间充质干细胞, H9C2细胞, 心力衰竭, 人参皂苷Rg1, 戊巴比妥钠, Toll样受体4, 核因子κB

Abstract: BACKGROUND: How to improve the expansion of cells, reduce cell loss, increase homing rate and reduce apoptosis is the main problem in the preclinical research of human umbilical cord mesenchymal stem cells. Ginsenoside Rg1 can promote the proliferation and differentiation of mesenchymal stem cells in different microenvironments in vitro or in vivo, which may be a candidate drug to improve the efficiency of human umbilical cord mesenchymal stem cell transplantation. 
OBJECTIVE: To investigate the effect of human umbilical cord mesenchymal stem cells co-culture combined with ginsenoside Rg1 on pentobarbital sodium induced heart failure cell model. 
METHODS: H9C2 cells were divided into five groups: Control group, pentobarbital sodium group, human umbilical cord mesenchymal stem cell group, ginsenoside Rg1 group, and human umbilical cord mesenchymal stem cell + ginsenoside Rg1 group. H9C2 cells in the control group were cultured in normal DMEM for 24 hours. H9C2 cells in the other groups were cultured in DMEM containing 0.8% pentobarbital sodium for 7 minutes to establish a heart failure cell model. After modeling, above models were treated with human umbilical cord mesenchymal stem cells, ginsenoside Rg1, or their combination. CCK-8 assay and EdU staining were used to detect cell proliferation. TUNEL assay was used to detect cell apoptosis. Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities were detected according to kit instructions. The mRNA levels of tumor necrosis factor α, interleukin 1β, and interleukin 6 in the supernatant were determined by ELISA. The mRNA levels of tumor necrosis factor α, interleukin 1β, interleukin 6, Bax, and Bcl2 in the cells were determined by RT-qPCR. The protein levels of Bax, Bcl2, Toll-like receptor 4, p65, and p-p65 were determined by western blot assay.  
RESULTS AND CONCLUSION: (1) Compared with the pentobarbital sodium group, H9C2 cell viability and EdU positive rate were increased; TUNEL positive rate and Bax mRNA and protein expression were decreased, and Bcl-2 mRNA and protein expression were increased; Na+-K+ -ATPase activity decreased; Ca2+-Mg2+-ATPase activity increased; tumor necrosis factor α, interleukin-1β, and interleukin-6 levels decreased in H9C2 cell supernatant, and tumor necrosis factor α, interleukin-1β, and interleukin-6 mRNA expression decreased in H9C2 cells; the expression of toll-like receptor 4 and P-P65 protein decreased with significant difference in human umbilical cord mesenchymal stem cell group, ginsenoside Rg1 group and human umbilical cord mesenchymal stem cell + ginsenoside Rg1 group (P < 0.05). (2) Compared with human umbilical cord mesenchymal stem cell group and ginsenoside Rg1 group, the above indexes in human umbilical cord mesenchymal stem cells + ginsenoside Rg1 group were further improved (P < 0.05). The results showed that human umbilical cord mesenchymal stem cells combined with ginsenoside Rg1 promoted the viability of heart failure cells induced by pentobarbital sodium and inhibited inflammation mediated by the Toll-like receptor 4/nuclear factor κB pathway.

Key words: human umbilical cord mesenchymal stem cell, H9C2 cell, heart failure, ginsenoside Rg1, pentobarbital sodium, Toll-like receptor 4, nuclear factor κB

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