Department of Ultrasound, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha, 410002, Hunan, China.
Department of Cardiovascular, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University, 61, Jiefang West Road, Furong District, Changsha City, 410002, Hunan, China.
Mol Biol Rep. 2022 Jul;49(7):6041-6052. doi: 10.1007/s11033-022-07392-3. Epub 2022 Mar 31.
Cardiomyocyte injury is a typical feature in cardiovascular diseases. Changes in cardiomyocytes strongly affect the progression of cardiovascular diseases. This work aimed to investigate the biological function and potential mechanism of action of miR-150-5p in cardiomyocytes.
A myocardial ischemia (MI) injury rat model was constructed to detect miR-150-5p and tetratricopeptide repeat domain 5 (TTC5) expression during heart ischemia injury. Primary cardiomyocytes were isolated for in vitro study. CCK-8 assays were used to detect cardiomyocyte viability. Western blots were used to detect TTC5 and P53 expression. qPCR was utilized to measure RNA expression of miR-150-5p and TTC5. The TUNEL assay was used to determine cell apoptosis. ELISA was used to determine cytokine (TNF-α, IL-1β, IL-6, and IL-8) levels in heart tissues and cell culture supernatants. A dual-luciferase reporter assay was carried out to verify the binding ability between miR-150-5p and TTC5. Oxygen-glucose deprivation (OGD) treatment significantly inhibited cell viability. Ultrasound-targeted microbubble destruction (UTMD)-mediated uptake of miR-150-5p inverted these results. Additionally, UTMD-mediated uptake of miR-150-5p retarded the effects of OGD treatment on cell apoptosis. Besides, UTMD-mediated uptake of miR-150-5p counteracted the effects of OGD treatment on the inflammatory response by regulating cytokine (TNF-α, IL-1β, IL-6, and IL-8) levels. For the mechanism of the protective effect on the heart, we predicted and confirmed that miR-150-5p bound to TTC5 and inhibited TTC5 expression.
UTMD-mediated uptake of miR-150-5p attenuated OGD-induced primary cardiomyocyte injury by inhibiting TTC5 expression. This discovery contributes toward further understanding the progression of primary cardiomyocyte injury.
心肌细胞损伤是心血管疾病的一个典型特征。心肌细胞的变化强烈影响心血管疾病的进展。本研究旨在探讨 miR-150-5p 在心肌细胞中的生物学功能和潜在作用机制。
构建心肌缺血（MI）损伤大鼠模型，检测心脏缺血损伤过程中 miR-150-5p 和四肽重复结构域 5（TTC5）的表达。分离原代心肌细胞进行体外研究。CCK-8 法检测心肌细胞活力。Western blot 法检测 TTC5 和 P53 表达。qPCR 法检测 miR-150-5p 和 TTC5 的 RNA 表达。TUNEL 法检测细胞凋亡。ELISA 法检测心脏组织和细胞培养上清液中细胞因子（TNF-α、IL-1β、IL-6 和 IL-8）水平。双荧光素酶报告实验验证 miR-150-5p 与 TTC5 的结合能力。氧葡萄糖剥夺（OGD）处理显著抑制细胞活力。超声靶向微泡破坏（UTMD）介导的 miR-150-5p 摄取逆转了这些结果。此外，UTMD 介导的 miR-150-5p 摄取通过调节细胞因子（TNF-α、IL-1β、IL-6 和 IL-8）水平，减缓 OGD 处理对细胞凋亡的影响。此外，UTMD 介导的 miR-150-5p 摄取通过调节细胞因子（TNF-α、IL-1β、IL-6 和 IL-8）水平，逆转 OGD 处理对炎症反应的影响。对于心脏的保护作用机制，我们预测并证实 miR-150-5p 与 TTC5 结合并抑制 TTC5 表达。
UTMD 介导的 miR-150-5p 摄取通过抑制 TTC5 表达减轻 OGD 诱导的原代心肌细胞损伤。这一发现有助于进一步了解原代心肌细胞损伤的进展。
