Department of Orthopedics, Changhai Hospital Affiliated to the Navy Military Medical University, 168 Changhai Road, Yangpu District 200433, Shanghai, China.
Department of Cardiology, Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine, 181 Friendship Road, Baoshan District 201901, Shanghai, China.
Int Immunopharmacol. 2022 Jun;107:108627. doi: 10.1016/j.intimp.2022.108627. Epub 2022 Feb 22.
This article aims to discuss the role of l KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1) in myocardial injury caused by a hip fracture and further investigate its potential molecular mechanisms.
X-Ray and H&E staining are used to observe hip fracture and pathological changes of myocardial tissue. ELISA and kits are used to detect inflammatory cytokines, lactate dehydrogenase (LDH), and creatine kinase (CK) in serum. The proliferation and apoptosis of H9c2 are determined by CCK-8 and flow cytometry. RT-qPCR and Western blot are applied to quantitatively assess the expression of related genes. Bioinformatics analysis is performed to search the downstream target of KCNQ1OT1 and miR-224-3p. Furthermore, the interaction is verified by a luciferase reporter assay.
A hip fracture model was successfully established. The high expression of inflammatory cytokines and cardiac injury markers indicated that hip fracture successfully induced myocardial injury. In TNF-ɑ treated cardiomyocyte model, high expression of KCNQ1OT1 promoted H9c2 cell proliferation and inhibited apoptosis. Furthermore, in the myocardial injury model rats induced by hip fracture, a high expression of KCNQ1OT1 reduced pathological damage in the myocardial tissue. Further research illustrated that miR-224-3p was the direct target of KCNQ1OT1, and GATA4 was the direct target of miR-224-3p. Importantly, functional research findings indicated that KCNQ1OT1 regulated myocardial injury caused by hip fracture via targeting the miR-224-3p/GATA4 axis.
Our study demonstrates that the KCNQ1OT1 suppresses myocardial injury via mediating miR-224-3p/GATA4, which provides a latent target for myocardial injury treatment.
本文旨在探讨长链非编码 RNA KCNQ1 反义转录本 1（KCNQ1OT1）在髋部骨折引起的心肌损伤中的作用，并进一步探讨其潜在的分子机制。
X 射线和 H&E 染色观察髋部骨折和心肌组织的病理变化。ELISA 和试剂盒检测血清中炎症细胞因子、乳酸脱氢酶（LDH）和肌酸激酶（CK）。CCK-8 和流式细胞术检测 H9c2 的增殖和凋亡。RT-qPCR 和 Western blot 定量评估相关基因的表达。生物信息学分析搜索 KCNQ1OT1 和 miR-224-3p 的下游靶基因。进一步通过荧光素酶报告基因实验验证其相互作用。
成功建立了髋部骨折模型。炎症细胞因子和心脏损伤标志物的高表达表明髋部骨折成功诱导了心肌损伤。在 TNF-ɑ 处理的心肌细胞模型中，KCNQ1OT1 的高表达促进了 H9c2 细胞的增殖并抑制了凋亡。此外，在髋部骨折诱导的心肌损伤模型大鼠中，KCNQ1OT1 的高表达减轻了心肌组织的病理损伤。进一步的研究表明，miR-224-3p 是 KCNQ1OT1 的直接靶基因，GATA4 是 miR-224-3p 的直接靶基因。重要的是，功能研究结果表明，KCNQ1OT1 通过靶向 miR-224-3p/GATA4 轴调节髋部骨折引起的心肌损伤。
本研究表明，KCNQ1OT1 通过介导 miR-224-3p/GATA4 抑制心肌损伤，为心肌损伤治疗提供了一个潜在的靶点。
