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1.075000 河北 张家口,河北北方学院基础医学院组织学与胚胎学教研室
2.075000 河北 张家口,河北北方学院干细胞与生殖生物学实验室
[王海萍,E-mail:2646077539@qq.com WANG Haiping, E-mail: 2646077539@qq.com
收稿:2024-04-03,
修回:2024-07-12,
纸质出版:2024-11-15
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董子晗, 王海萍, 吕洋, 等. FGF-2和三七总皂苷诱导骨髓间充质干细胞向心肌样细胞分化[J]. 陆军军医大学学报, 2024,46(21):2415-2423.
Zihan DONG, Haiping WANG, Yang LYU, et al. Differentiation of bone marrow mesenchymal stem cells into cardiomyocytes with FGF-2 and Panax notoginseng saponins[J]. Journal of Army Medical University, 2024, 46(21): 2415-2423.
董子晗, 王海萍, 吕洋, 等. FGF-2和三七总皂苷诱导骨髓间充质干细胞向心肌样细胞分化[J]. 陆军军医大学学报, 2024,46(21):2415-2423. DOI: 10.16016/j.2097-0927.202404018.
Zihan DONG, Haiping WANG, Yang LYU, et al. Differentiation of bone marrow mesenchymal stem cells into cardiomyocytes with FGF-2 and Panax notoginseng saponins[J]. Journal of Army Medical University, 2024, 46(21): 2415-2423. DOI: 10.16016/j.2097-0927.202404018.
目的
2
探究碱性成纤维细胞生长因子(basic fibro-blast growth factor
FGF-2)和三七总皂苷(Panax notoginseng saponins,PNS)单独或联合诱导对骨髓间充质干细胞(bone mesenchymal stem cells
BMSCs)向心肌细胞(cardiomyocytes
CMs)分化的影响以及PNS和FGF-2促分化的作用机制。
方法
2
全骨髓贴壁法分离20~30 g 3周龄SD大鼠的骨髓间充质干细胞,通过流式细胞术鉴定骨髓间充质干细胞的纯度。鉴定纯度合格后通过PNS和FGF-2单独或联合诱导以及PNS和FGF-2分别与LY294002(phosphatidylinositol 3 kinase抑制剂)联用诱导骨髓间充质干细胞向心肌细胞分化。根据培养的相应时间点,分为对照组、三七总皂苷组(PNS组)、碱性成纤维细胞生长因子组(FGF-2组)、三七总皂苷+碱性成纤维细胞生长因子组(PNS+FGF-2组)。通过免疫组织化学检测诱导后Desmin和Cx43的表达水平;Western blot检测心肌特异性蛋白、有关通路蛋白及细胞凋亡有关蛋白的表达水平;采用RT-qPCR检测心肌转录因子表达水平。
结果
2
各单独诱导组(FGF-2组、PNS组)或联合诱导组(PNS+FGF-2组)均提高心肌特异性蛋白和基因表达水平(
P
<0.05),且最高表达水平出现在联合组。PNS和FGF-2分别与LY294002联用,相较于两单独诱导组降低了心肌特异性蛋白和基因的表达水平,降低通路相关蛋白Akt的磷酸化水平,且Bcl-2水平降低,Bax水平升高,Bcl-2/Bax比值减小(
P
<0.05)。
结论
2
PNS和FGF-2均可使骨髓间充质干细胞诱导后向心肌细胞定向分化,且二者的联合诱导是一种更高效的诱导方法。PI3K/Akt信号通路参与了PNS和FGF-2促进骨髓间充质干细胞向心肌细胞分化的过程,且PNS和FGF-2起到减少细胞凋亡的作用。
Objective
2
To investigate the effects of basic fibro-blast growth factor (FGF-2) and Panax notoginseng saponins (PNS) alone or combined together on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocytes (CMs) and the mechanism of action of PNS and FGF-2 in the differentiation.
Methods
2
BMSCs were isolated from 3-week-old SD rats (20~30 g) by whole bone marrow adherence
and then the purity of BMSCs was identified by flow cytometry. After the purity was qualified
PNS and FGF-2 alone or in combination
or combined with LY294002 (phosphatidylinositol 3 kinase inhibitor)
were used to induce BMSCs to differentiate. According to the corresponding culture time
the cells were divided into control group
PNS group
FGF-2 group
and PNS+FGF-2 group. The expression levels of Desmin and Cx43 after induction were detected by immunohistochemical assay. Western blotting was used to measure the expression levels of myocardium-specific proteins
related pathway proteins
and apoptosis-related proteins. RT-qPCR was conducted to detect the expression of myocardial transcription factors.
Results
2
The expression levels of myocardium-specific proteins and genes were increased in each single induction group (PNS group and FGF-2 group) and combined induction group (PNS+FGF-2 group
P
<
0.05)
and the highest expression levels were observed in the combined group. PNS and FGF-2 combined with LY294002 decreased the expression levels of myocardium-specific proteins and genes
decreased the phosphorylation of pathway-related protein Akt
decreased the level of Bcl-2
increased the level of Bax
and decreased the ratio of Bcl-2/Bax when compared with the 2 single induction groups (
P
<
0.05).
Conclusion
2
Both PNS and FGF-2 can induce BMSCs to differentiate into CMs after induction
and the combined induction is more effective in the differentiation. The PI3K/Akt signaling pathway is involved in the differentiation process by PNS and FGF-2
and PNS and FGF-2 play a role in reducing apoptosis.
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