中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (7): 1032-1036.doi: 10.3969/j.issn.2095-4344.2168

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

丹酚酸B抑制骨髓间充质干细胞氧化损伤及促进分化为心肌样细胞

裴丽丽1，孙贵才2，王  弟3   

  1. 1秦皇岛中医医院，河北省秦皇岛市   066000；2南昌大学第一附属医院骨科，江西省南昌市   330003；3齐齐哈尔医学院附属第一医院，黑龙江省齐齐哈尔市   161041
  • 收稿日期:2019-09-05 修回日期:2019-09-07 接受日期:2020-03-04 出版日期:2021-03-08 发布日期:2020-12-08
  • 通讯作者: 孙贵才，主任中医师，南昌大学第一附属医院骨科，江西省南昌市 330003
  • 作者简介:裴丽丽，女，1982年生，黑龙江省齐齐哈尔市人，汉族，2009年吉林大学毕业，硕士，主治医师。
  • 基金资助:
    国家自然科学基金项目(81960881，81660801)

Salvianolic acid B inhibits oxidative damage of bone marrow mesenchymal stem cells and promotes differentiation into cardiomyocytes

Pei Lili1, Sun Guicai2, Wang Di3   

  1. 1Qinhuangdao Hospital of Traditional Chinese Medicine, Qinhuangdao 066000, Hebei Province, China; 2Department of Orthopedics, First Affiliated Hospital of Nanchang University, Nanchang 330003, Jiangxi Province, China; 3First Affiliated Hospital of Qiqihar Medical University, Qiqihar 161041, Heilongjiang Province, China
  • Received:2019-09-05 Revised:2019-09-07 Accepted:2020-03-04 Online:2021-03-08 Published:2020-12-08
  • Contact: Sun Guicai, Chief TCM physician, Department of Orthopedics, First Affiliated Hospital of Nanchang University, Nanchang 330003, Jiangxi Province, China
  • About author:Pei Lili, Master, Attending physician, Qinhuangdao Hospital of Traditional Chinese Medicine, Qinhuangdao 066000, Hebei Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81960881, 81660801

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摘要：

文题释义：
Nrf2：Nrf2与细胞质中的Keap1结合，导致蛋白酶体降解。在应激暴露时，出现Keap1半胱氨酸修饰的特定模式。Keap1释放 Nrf2，其转移到细胞核后与小Maf 蛋白形成异二聚体复合物。该复合物识别位于细胞保护基因中的抗氧化反应元件的增强子序列，激活它的转录。
Keap1：是由624个氨基酸残基组成的二聚体蛋白质，主要位于细胞质中，Keap1的特定半胱氨酸残基修饰是氧化应激条件下细胞中蛋白质构象变化的原因，Keap1 可与Nrf2结合阻止其进入细胞核。

背景：近年来，骨髓间充质干细胞已用于临床治疗由永久性心肌细胞减少引起的心脏损伤。然而，促进骨髓间充质干细胞增殖及向心肌细胞分化的最佳诱导剂仍然在研究筛选当中。 
目的：探讨丹酚酸B对骨髓间充质干细胞氧化应激损伤的保护作用及向心肌样细胞分化的影响。
方法：以骨髓间充质干细胞为研究对象，用300 μmol/L过氧化氢干预24 h建立氧化应激损伤模型，同时给予丹酚酸B干预24 h，采用试剂盒检测各组细胞上清液中乳酸脱氢酶和肌酸激酶水平及细胞裂解液中丙二醛和超氧化物歧化酶水平，Western blot检测骨髓间充质干细胞中Nrf2、Keap1蛋白与Bcl-2、Bax蛋白表达，干预14 d后采用RT-PCT检测心肌细胞分化相关标记物的表达。
结果与结论：①与模型组比较，丹酚酸B组上清液中肌酸激酶和乳酸脱氢酶水平明显降低(P < 0.05)；②与模型组比较，丹酚酸B组细胞裂解液中丙二醛水平降低，超氧化物歧化酶水平升高(P < 0.05)；③与模型组比较，丹酚酸B组细胞中Nrf2蛋白表达明显增加，Keap1蛋白表达显著降低；④与模型组比较，丹酚酸B组Bcl-2蛋白表达显著提高，Bax蛋白表达显著降低；⑤与模型组比较，丹酚酸B组GATA4、cTnT mRNA表达显著提高；⑥结果表明丹酚酸B能够诱导骨髓间充质干细胞激活Nrf2-ARE信号通路相关蛋白的表达从而达到抗氧化作用，抑制细胞凋亡，还可以提高骨髓间充质干细胞向心肌样细胞分化的能力。
https://orcid.org/0000-0002-1515-9378(孙贵才) 

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: 干细胞, 骨髓间充质干细胞, 丹酚酸B, 心肌细胞, 氧化应激, 凋亡, 通路

Abstract: BACKGROUND: In recent years, bone marrow mesenchymal stem cells have been used in clinical treatment of heart injury caused by permanent cardiomyocytopenia. However, the best inducer to promote the proliferation and differentiation of bone marrow mesenchymal stem cells into cardiomyocytes is still in the research and selection.
OBJECTIVE: To investigate the protective effect of salvianolic acid B on oxidative stress injury and cardiomyocyte differentiation of bone marrow mesenchymal stem cells. 
METHODS:  Bone marrow mesenchymal stem cells were used as study objects to establish oxidative stress injury model by 300 μmol/L H2O2 for 24 hours. Simultaneously, bone marrow mesenchymal stem cells were treated with salvianolic acid B for 24 hours. The contents of lactate dehydrogenase and creatine kinase in the supernatant and malondialdehyde and superoxide dismutase in the cell lysate were determined in each group using the kit. Western blot assay was used to detect the expression of nuclear factor E2 related factor 2 (Nrf2), Keap1, Bcl-2 and Bax in bone marrow mesenchymal stem cells. RT-PCT was used to detect cardiomyocyte differentiation-related markers 14 days after intervention.  
RESULTS AND CONCLUSION: (1) Compared with the model group, the levels of lactate dehydrogenase and creatine kinase in the supernatant were significantly decreased in the salvianolic acid B group (P < 0.05). (2) Compared with the model group, the level of malondialdehyde in cell lysate was decreased, and the level of superoxide dismutase was increased in the salvianolic acid B group (P < 0.05). (3) Compared with the model group, expression of Nrf2 protein in cells was significantly increased, and the expression of Keap1 protein was significantly decreased in the salvianolic acid B group. (4) The expression of Bcl-2 in salvianolic acid B group was significantly higher than that in model group, and the expression of Bax in salvianolic acid B group was significantly lower than that in model group. (5) Compared with the model group, the expression of GATA4 and cTnT mRNA was significantly increased in the salvianolic acid B group. (6) Salvianolic acid B can induce the expression of proteins that activate Nrf2-ARE signaling pathway in bone marrow mesenchymal stem cells for anti-oxidation, and salvianolic acid B can inhibit apoptosis and improve the ability of bone marrow mesenchymal stem cells to differentiate into cardiomyocytes.

Key words: stem cells, bone marrow mesenchymal stem cells, salvianolic acid B, cardiomyocytes, oxidative stress, apoptosis, pathway

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