Xin Zhong1, Yu Chen2, Xiangdang Long1, Hongtian Chen1, Zhaofen Zheng2, Hongwei Pan2, Jianqiang Peng2, Yanfu Liu3, Haijun Wang3, Yongjun Hu4. 1. Department of Ultrasound, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University, Changsha, 410002, Hunan, China. 2. 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. 3. Department of Cardiovascular, Lixian People's Hospital, Changde, 415500, Hunan, China. 4. 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. kuqsdi@163.com.
Abstract
BACKGROUND: 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. METHODS AND RESULTS: 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. CONCLUSIONS: 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.
BACKGROUND: 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. METHODS AND RESULTS: 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. CONCLUSIONS: 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.
Authors: Robin Verjans; Tim Peters; Francisco Javier Beaumont; Rick van Leeuwen; Tessa van Herwaarden; Wouter Verhesen; Chantal Munts; Mitchell Bijnen; Michiel Henkens; Javier Diez; Leon J de Windt; Frans A van Nieuwenhoven; Marc van Bilsen; Marie José Goumans; Stephane Heymans; Arantxa González; Blanche Schroen Journal: Hypertension Date: 2017-12-18 Impact factor: 10.190
Authors: Mitchell C Lock; Ross L Tellam; Kimberley J Botting; Kimberley C W Wang; Joseph B Selvanayagam; Doug A Brooks; Mike Seed; Janna L Morrison Journal: J Physiol Date: 2018-06-28 Impact factor: 5.182