ABC | Volume 111, Nº2, August 2018

Original Article Zhong et al Pioglitazone and VEGFR-2 signaling pathway Arq Bras Cardiol. 2018; 111(2):162-169 Figure 3 – Pioglitazone and Apatinib regulate VEGFR-2 signaling in neonatal rat cardiomyocytes under hypertrophy induced by Angiotensin II (n = 12 in each group). A and B, Representative western bolotting trace of phospho-VEGFR-2, VEGFR-2, phospho-mTOR, mTOR, phospho-Akt, Akt, Bax, phospho-P53 and P53 protein levels in rat neonatal cardiomyocytes under hypertrophic stimuli, and treated with pioglitazone (20 μM) or apatinib (2 μM) for 24 hours, and intensity of the above bands in A normalized to GAPDH. All data represent the means ± SD. *p < 0.01 compared with controls; #p < 0.01 compared withi pioglitazone 20 μM group, calculated by one-way ANOVA followed by the post hoc Bonferroni test for pairwise comparisons. Control Pioglitazone 20 µM Apatinib 2 µM Control Pioglitazone 20 µM Apatinib 2 µM Control Pioglitazone 20 µM Apatinib 2 µM Control Pioglitazone 20 µM Apatinib 2 µM Control Pioglitazone 20 µM Apatinib 2 µM Control Pioglitazone 20 µM Apatinib 2 µM A B pVEGFR2 (Tyr1175) VEGFR2 pmTOR (Ser2448) mTOR pAkt (Thr308) Akt bax pP53(Ser15) P53 GAPDH Normalized pVEGFR2 (Tyr1175) levels Relative Bax protein expression Normatized pP53(Sr15) levels Normatized pAKT (Thr308) levels Normatized pmTOR (Ser2448) levels 1.5 0.5 1.0 0.0 1.5 0.5 1.0 0.0 1.5 0.5 1.0 0.0 4 3 2 1 0 4 3 2 1 0 * * * * * *# *# *# *# *# also participates in survival and hypertrophy of these cells by inhibiting P53-dependent pathways and activating mTOR-dependent pathways, respectively. 37 In this study, pioglitazone and VEGFR-2 inhibitor apatinib increased phospho-P53 and Bax expression in cardiomyocytes and decreased phospho-Akt and phospho-mTOR expression in hypertrophic cardiomyocytes, indicating the connection between pioglitazone and the VEGFR-2 signaling pathway. Conclusion In conclusion, these findings indicate that pioglitazone induces apoptosis and inhibits hypertrophy of cardiomyocytes in part by acting on the VEGFR-2 signaling pathway. These findings contribute to understanding cardiovascular risks of pioglitazone. Author contributions Conception and design of the research: Zhong W, Chen L; Acquisition of data: Zhong W, Jin W, Wu Y, Luo S, Liang M; Analysis and interpretation of the data and statistical analysis: Zhong W, Jin W, Xu S, Wu Y, Luo S, Liang M;Obtaining financing: Zhong W; Writing of the manuscript: Zhong W, Jin W, Liang M; Critical revision of the manuscript for intellectual content: Zhong W, Xu S, Liang M. Potential Conflict of Interest No potential conflict of interest relevant to this article was reported. Sources of Funding This study was funded by the Natural Science Foundation of Fujian Province Project (N°. 2010J01371). Study Association This article is part of the thesis of Post-Doctoral submitted by WenliangZhong, fromUnionHospital of FujianMedical University. Ethics approval and consent to participate This study was approved by the Ethics Committee on Animal Experiments of the First Hospital of Nanping City under the protocol number NP01371. 167