Effect of AMP-activated protein kinase activation on cardiac fibroblast proliferation induced by coxsackievirus B3.
Excessive fibroblast proliferation and collagen production are the major pathogenic mechanisms in the progression of viral myocarditis, which is most frequently associated with infection by coxsackievirus B3 (CVB3). AMP-activated protein kinase (AMPK) has been confirmed to be involved in the progression of myocardial remodeling. However, it remains unclear whether AMPK has an effect on CVB3-induced cardiac fibroblast proliferation. In the present study, the effects of AMPK on cardiac fibroblast proliferation and collagen secretion induced by CVB3 were investigated. Proliferation of neonatal cardiac fibroblasts was determined by cell counting and detection of newly synthesized DNA in cells, and the proportion of cells in the S phase was measured. Hydroxyproline ELISA was used to detect collagen secretion. Phosphorylation of AMPKα-Thr172was evaluated by western blotting. It was found that neonatal cardiac fibroblasts were clearly proliferating markedly and secreting collagen at 24 h after CVB3 infection, and peaked at 48 h. These effects were inhibited following pretreatment of the fibroblasts with 5-aminoimidazole-4-carboxamide-ribonucleoside (AICAR), a specific AMPK activator, for 2 h prior to CVB3 infection. However, if the cells were preincubated with compound C for 30 min, the inhibitive effects of AICAR were reversed. Western blotting results indicated that AMPK α-Thr172phosphorylation was increased by AICAR and attenuated by Compound C. The results of the present study suggest that CVB3 infection increases cardiac fibroblast proliferation and collagen secretion, and that these phenomena can be inhibited by activated AMPK. These findings contribute to our understanding of AMPK function and the future design of therapeutic approaches for the treatment of cardiac fibrosis caused by chronic viral infection, such as CVB3-induced myocarditis.
Department of Cardiology, The Third Affiliated Hospital of Nantong University, Wuxi Institute of Integrated Traditional Chinese and Western Medicine, Wuxi, Jiangsu 214041, P.R. China.
Shengyang
S
Jiang
Department of Geratology, Tianjin Geriatric Institute, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China.
Shunli
S
Tian
Department of Cardiology, The Third Affiliated Hospital of Nantong University, Wuxi Institute of Integrated Traditional Chinese and Western Medicine, Wuxi, Jiangsu 214041, P.R. China.
Xueming
X
Wu
Department of Cardiology, The Third Affiliated Hospital of Nantong University, Wuxi Institute of Integrated Traditional Chinese and Western Medicine, Wuxi, Jiangsu 214041, P.R. China.
Yijia
Y
Tao
Clinical Central Laboratory, The Third Affiliated Hospital of Nantong University, Wuxi Institute of Integrated Traditional Chinese and Western Medicine, Wuxi, Jiangsu 214041, P.R. China.
Donglin
D
Jiang
AMP-activated protein kinase; cardiac fibroblast; cell proliferation; collagen; coxsackievirus