Dr. Zafar H. Zaidi Center for Proteomics, University of Karachi, Karachi, 75270, Pakistan.
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan.
Mol Biol Rep. 2023 Sep;50(9):7371-7380. doi: 10.1007/s11033-023-08634-8. Epub 2023 Jul 14.
Cardiovascular diseases remain a major cause of death globally. Cardiac cells once damaged, cannot resume the normal functioning of the heart. Bone marrow derived mesenchymal stem cells (BM-MSCs) have shown the potential to differentiate into cardiac cells. Epigenetic modifications determine cell identity during embryo development via regulation of tissue specific gene expression. The major epigenetic mechanisms that control cell fate and biological functions are DNA methylation and histone modifications. However, epigenetic modifiers alone are not sufficient to generate mature cardiac cells. Various small molecules such as ascorbic acid (AA) and salvianolic acid B (SA) are known for their cardiomyogenic potential. Therefore, this study is aimed to examine the synergistic effects of epigenetic modifiers, valproic acid (VPA) and 5-azacytidine (5-aza) with cardiomyogenic molecules, AA and SA in the cardiac differentiation of MSCs.
BM-MSCs were isolated, propagated, characterized, and then treated with an optimized dose of VPA or 5-aza for 24 h. MSCs were maintained in a medium containing AA and SA for 21 days. All groups were assessed for the expression of cardiac genes and proteins through q-PCR and immunocytochemistry, respectively. Results show that epigenetic modifiers VPA or 5-aza in combination with AA and SA significantly upregulate the expression of cardiac genes MEF2C, Nkx2.5, cMHC, Tbx20, and GATA-4. In addition, VPA or 5-aza pretreatment along with AA and SA enhanced the expression of the cardiac proteins connexin-43, GATA-4, cTnI, and Nkx2.5.
These findings suggest that epigenetic modifiers valproic acid and 5-azacytidine in combination with ascorbic acid and salvianolic acid B promote cardiac differentiation of MSCs. This pretreatment strategy can be exploited for designing future stem cell based therapeutic strategies for cardiovascular diseases.
心血管疾病仍然是全球主要的死亡原因。心脏细胞一旦受损，就无法恢复心脏的正常功能。骨髓间充质干细胞（BM-MSCs）已显示出分化为心脏细胞的潜力。表观遗传修饰通过调节组织特异性基因表达来决定胚胎发育过程中的细胞身份。控制细胞命运和生物学功能的主要表观遗传机制是 DNA 甲基化和组蛋白修饰。然而，单独的表观遗传修饰剂不足以产生成熟的心脏细胞。已知各种小分子，如抗坏血酸（AA）和丹酚酸 B（SA）具有心脏生成潜力。因此，本研究旨在研究表观遗传修饰剂丙戊酸（VPA）和 5-氮杂胞苷（5-aza）与心脏生成分子 AA 和 SA 协同作用在 MSC 心脏分化中的作用。
分离、增殖、鉴定 BM-MSCs，然后用优化剂量的 VPA 或 5-aza 处理 24 小时。将 MSC 维持在含有 AA 和 SA 的培养基中 21 天。通过 q-PCR 和免疫细胞化学分别评估所有组的心脏基因和蛋白质的表达。结果表明，表观遗传修饰剂 VPA 或 5-aza 与 AA 和 SA 联合使用可显著上调 MEF2C、Nkx2.5、cMHC、Tbx20 和 GATA-4 等心脏基因的表达。此外，VPA 或 5-aza 预处理与 AA 和 SA 联合使用可增强连接蛋白 43、GATA-4、cTnI 和 Nkx2.5 等心脏蛋白的表达。
这些发现表明，表观遗传修饰剂丙戊酸和 5-氮杂胞苷与抗坏血酸和丹酚酸 B 联合使用可促进 MSC 的心脏分化。这种预处理策略可用于设计未来基于干细胞的心血管疾病治疗策略。
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