长链非编码 RNA-心脏诱导 RNA6 通过细胞周期蛋白依赖性激酶 1 诱导间充质干细胞向成心肌细胞分化来介导梗死心脏的修复。

Long Non-Coding RNA-Cardiac-Inducing RNA 6 Mediates Repair of Infarcted Hearts by Inducing Mesenchymal Stem Cell Differentiation into Cardiogenic Cells through Cyclin-Dependent Kinase 1.

机构信息

Department of Physiology and Pathophysiology, College of Medicine, Yanbian University, Yanji 133002, China.

出版信息

Int J Mol Sci. 2024 Mar 19;25(6):3466. doi: 10.3390/ijms25063466.

DOI:10.3390/ijms25063466

PMID:38542439

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10971150/

Abstract

This study aims to investigate the induction effect of LncRNA-CIR6 on MSC differentiation into cardiogenic cells in vitro and in vivo. In addition to pretreatment with Ro-3306 (a CDK1 inhibitor), LncRNA-CIR6 was transfected into BMSCs and hUCMSCs using jetPRIME. LncRNA-CIR6 was further transfected into the hearts of C57BL/6 mice via 100 μL of AAV9-cTnT-LncRNA-CIR6-ZsGreen intravenous injection. After three weeks of transfection followed by AMI surgery, hUCMSCs (5 × 10/100 μL) were injected intravenously one week later. Cardiac function was evaluated using VEVO 2100 and electric mapping nine days after cell injection. Immunofluorescence, Evans blue-TTC, Masson staining, FACS, and Western blotting were employed to determine relevant indicators. LncRNA-CIR6 induced a significant percentage of differentiation in BMSCs (83.00 ± 0.58)% and hUCMSCs (95.43 ± 2.13)% into cardiogenic cells, as determined by the expression of cTnT using immunofluorescence and FACS. High cTNT expression was observed in MSCs after transfection with LncRNA-CIR6 by Western blotting. Compared with the MI group, cardiac contraction and conduction function in MI hearts treated with LncRNA-CIR6 or combined with MSCs injection groups were significantly increased, and the areas of MI and fibrosis were significantly lower. The transcriptional expression region of LncRNA-CIR6 was on Chr17 from 80209290 to 80209536. The functional region of LncRNA-CIR6 was located at nucleotides 0-50/190-255 in the sequence. CDK1, a protein found to be related to the proliferation and differentiation of cardiomyocytes, was located in the functional region of the LncRNA-CIR6 secondary structure (from 0 to 17). Ro-3306 impeded the differentiation of MSCs into cardiogenic cells, while MSCs transfected with LncRNA-CIR6 showed a high expression of CDK1. LncRNA-CIR6 mediates the repair of infarcted hearts by inducing MSC differentiation into cardiogenic cells through CDK1.

摘要

本研究旨在探讨长链非编码 RNA-CIR6 在体外和体内诱导间充质干细胞向心肌细胞分化的诱导作用。除了用 Ro-3306（CDK1 抑制剂）预处理外，还通过 jetPRIME 将 LncRNA-CIR6 转染到 BMSCs 和 hUCMSCs 中。进一步通过静脉注射 100μL 的 AAV9-cTnT-LncRNA-CIR6-ZsGreen 将 LncRNA-CIR6 转染到 C57BL/6 小鼠的心脏中。转染 3 周后进行 AMI 手术，1 周后静脉注射 5×10/100μL 的 hUCMSCs。细胞注射后 9 天，使用 VEVO 2100 和电映射评估心功能。通过免疫荧光、Evans 蓝-TTC、Masson 染色、FACS 和 Western blot 测定相关指标。免疫荧光和 FACS 检测 cTnT 的表达表明，LncRNA-CIR6 诱导 BMSCs（83.00±0.58）%和 hUCMSCs（95.43±2.13）%分化为心肌细胞的比例显著增加。Western blot 检测到 LncRNA-CIR6 转染后的 MSCs 中 cTNT 表达高。与 MI 组相比，LncRNA-CIR6 或联合 MSC 注射组治疗的 MI 心脏的心脏收缩和传导功能明显增加，MI 和纤维化面积明显降低。LncRNA-CIR6 的转录表达区域位于 Chr17 上，从 80209290 到 80209536。LncRNA-CIR6 的功能区域位于序列中的 0-50/190-255 核苷酸。CDK1 是一种与心肌细胞增殖和分化相关的蛋白，位于 LncRNA-CIR6 二级结构的功能区域（0-17）。Ro-3306 阻碍 MSCs 向心肌细胞分化，而转染 LncRNA-CIR6 的 MSCs 中 CDK1 表达较高。LncRNA-CIR6 通过诱导 MSC 分化为心肌细胞来介导受损心脏的修复。