Author:
Ouyang Weiwei,Fu Shimei,Zhao Xing,Su Shengfa,Zhang Jun,Luo Daxian,Liu Lina,Ding Wenjin,Cao Dongdong,Liu Likun,He Zhixu,Lu Bing
Abstract
Abstract
Purpose
The aim of the present study was to investigate the efficacy of recombinant human endostatin (ES) (rh-ES) combined with radiation on rat cardiomyocyte apoptosis and the regulatory mechanism of transforming growth factor beta1 (TGF-β1)/Sma and Mad-related protein 3 (Smad3)/connective tissue growth factor (CTGF) signaling.
Method
The primary cardiomyocytes were isolated from neonatal Sprague–Dawley rats for culture in vitro and divided into blank control group (without treatment), 10 Gy radiation + siTGF-β1 siRNA (gene silencing) group, ES + siTGF-β1 siRNA group, and 10 Gy radiation + ES + siTGF-β1 siRNA group. Methyl thiazolyl tetrazolium assay was used to calculate the half-maximal inhibitory concentration (IC50) of rh-ES on cardiomyocytes. Adenoviral vector was constructed for virus packaging to silence TGF-β1 expression in cardiomyocytes. Quantitative real-time polymerase chain reaction and Western blot were carried out to analyze TGF-β1, Smad2, Smad3 and CTGF expression at both gene and protein levels. Flow cytometry and electron microscope were used to examine cell apoptosis.
Results
ES had a dose-dependent inhibitory effect on the proliferation of primary rat cardiomyocytes. ES combined with radiotherapy significantly inhibited cardiomyocyte proliferation and promoted cell apoptosis (P < 0.01). The gene and protein expression of TGF-β1, Smad2, Smad3 and CTGF were significantly up-regulated in primary cardiomyocytes transfected with TGF-β1 gene (P < 0.05).
Conclusion
The combination therapy with rh-ES and radiation can promote cardiomyocyte apoptosis and aggravate myocardial cell damage via TGF-β1/Smad3/CTGF signaling pathway.
Funder
National Science Foundation of China
Project of the Science and Technology Department of Guizhou Province
Guizhou Province Application of Basic Research Program Major Projects
Guizhou Provincial Department of Education Innovation Group Research Project
China International Medical Exchange Foundation
The Affiliated Hospital of Guizhou Medical University and the Affiliated Cancer Hospital of Guizhou Medical University
Publisher
Springer Science and Business Media LLC
Subject
Cardiology and Cardiovascular Medicine
Cited by
3 articles.
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