Cilostazol ameliorates heart failure with preserved ejection fraction and diastolic dysfunction in obese and non-obese hypertensive mice
Authors: Manoj Kumar Barthwal
Heena Agarwal
Sukka Santosh Reddy
Journal: Journal of Molecular and Cellular Cardiology (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: AngiotensinII; Cilostazol; Diastolic dysfunction; HFpEF; Obesity
Affiliations: Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific & Innovative Research (AcSIR), New Delhi 110025, India., Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow 226031, India., Pharmacology Division, CSIR-Central Drug Research Institute, Lucknow 226031, India. Electronic address: .

Abstract:  Cilostazol (Ciloz) a potent Type III phosphodiesterase inhibitor is effective against inflammation, insulin resistance and cardiomyopathy. However, the effect of Ciloz on obesity-associated left ventricular diastolic dysfunction has not been explored yet. Hence, we examined the effect of Ciloz on cardiac remodelling and dysfunction in non-obese and obese-insulin resistant mice infused with AngiotensinII (AngII). Male C57BL/6 J mice were initially subjected to 19 weeks of chow or high fat diet (HFD) regimen and thereafter animals were randomised for AngII (1500 ng/kg/min, s.c) infusion or saline and Ciloz (50 mg/kg, p.o) for another 1 week. Obese and non-obese mice infused with AngII exhibited significant diastolic dysfunction and features of heart failure with preserved ejection fraction (HFpEF) since a decrease in fractional shortening and no change in ejection fraction were observed when compared with respective controls. Administration of AngII and Ciloz in HFD fed mice significantly improved the left ventricular function compared with AngII infused HFD mice as evinced from the echocardiographic data. Further, Ciloz treatment significantly reduced cardiomyocyte area, interstitial and perivascular fibrosis; and collagen deposition. Moreover, Ciloz reduced the inflammatory milieu in the heart as evinced by decreased F4/80+ and CD68+ cells; IL-1β and IL-6 gene transcripts. Quantitative assessment of the expression levels revealed substantial upregulation of MMP-9 (pro- and mature-forms) and α-SMA in the left ventricle of AngII infused HFD-fed mice, which was considerably suppressed by Ciloz regimen. The beneficial effect of Ciloz was associated with the normalization in gene expression of hypertrophic and fibrotic markers. Likewise, Ciloz administration markedly reduced the AngII and HFD induced TGF-β1/SMAD3 and Akt/mTOR signalling. Additionally, AngII administered and HFD-fed mice showed increased glycolytic flux, which was considerably diminished by Ciloz treatment as indicated from suppressed PKM2, HK-2, PDK-2, HIF-1α mRNA and GLUT-1 protein expression. Taken together, Ciloz might be therapeutically exploited against AngII and obesity-associated diastolic dysfunction thereby preventing overt heart failure. 

Retracted: No

PubPeer comments:
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 For E-H: Thanks for your comments. To support our results, we will upload the whole blots very soon. However, it has to be understood that since the common whole cell lysates from different treatment groups have been blotted for the expression of different proteins, the actin expression and profile will be the same. The results depicted in panel H I are corroborated by the results shown in panel B and C respectively. The phosphorylation results shown in E, G are also supported by the expression of total proteins in the same panel. Therefore, there is no bearing on outcome of the study. We appreciate your concern and interest in our work. For I: Appreciate the comments and concern. I agree with the comment that there is background and some non-specificity signals are also present which is not unusual when doing Westerns with varied antibodies available in the market. Sometimes polyclonal and ascites antibodies do give lot of backgrounds. Sometimes multiple bands may also be due to presence of splice variants, degradation or post-translationally modified products of the original protein. Specificity of the band is ascertained on the basis of molecular weight and previous reported literature under similar conditions, like done in the present case also. Where there is less information characterization of the protein in the given cell, we use competing peptides to ascertain the signal specificity. Initially most of the time a positive control is also, run to determine the specificity of antibody. Therefore, we take a holistic and situation dependent case specific approach for ascertaining such issues and utmost care is taken. Yes, background issues are there with the antibody, just like with many other ones. Besides this, the results shown in this panel are also supported and confirmed by data presented in panel C by different methodology. Therefore, the present scenario does not affect the outcome of the study. Apologies for any inconvenience. ,