Cardiac Protective Effect of Kirenol against Doxorubicin-Induced Cardiac Hypertrophy in H9c2 Cells through Nrf2 Signaling via PI3K/AKT Pathways
Authors: Hamza Hanieh
Vishnu Priya Veeraraghavan
Abdullah M Alzahrani
Peramaiyan Rajendran
Journal: International Journal of Molecular Sciences (2021)
Discipline: CHEMISTRY
Keywords: H9c2; Nrf2; cardiotoxicity; doxorubicin; hypertrophy
Affiliations: Department of Biological Sciences, College of Science, King Faisal University, Al Ahsa 31982, Saudi Arabia., Department of Biochemistry, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai 600 077, India., Department of Medical Analysis, Al-Hussein Bin Talal University, Ma'an 71111, Jordan.

Abstract:  Kirenol (KRL) is a biologically active substance extracted from Herba Siegesbeckiae. This natural type of diterpenoid has been widely adopted for its important anti-inflammatory and anti-rheumatic properties. Despite several studies claiming the benefits of KRL, its cardiac effects have not yet been clarified. Cardiotoxicity remains a key concern associated with the long-term administration of doxorubicin (DOX). The generation of reactive oxygen species (ROS) causes oxidative stress, significantly contributing to DOX-induced cardiac damage. The purpose of the current study is to investigate the cardio-protective effects of KRL against apoptosis in H9c2 cells induced by DOX. The analysis of cellular apoptosis was performed using the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining assay and measuring the modulation in the expression levels of proteins involved in apoptosis and Nrf2 signaling, the oxidative stress markers. Furthermore, Western blotting was used to determine cell survival. KRL treatment, with Nrf2 upregulation and activation, accompanied by activation of PI3K/AKT, could prevent the administration of DOX to induce cardiac oxidative stress, remodeling, and other effects. Additionally, the diterpenoid enhanced the activation of Bcl2 and Bcl-xL, while suppressing apoptosis marker proteins. As a result, KRL is considered a potential agent against hypertrophy resulting from cardiac deterioration. The study results show that KRL not only activates the IGF-IR-dependent p-PI3K/p-AKT and Nrf2 signaling pathway, but also suppresses caspase-dependent apoptosis. 

Retracted: No

PubPeer comments:
 These two data may be identical.![file] , 
 They may use same beta-actin data as internal controls in different experiments.![file] , 
 180 degrees rotation may provide different data. ![file] , 
 The bands in Fig 3B unnaturally look similar. ![file] , 
 Ok Mr Acrostalagmus annulatus, I think you came on us very viciously with a bad intent. You started throwing different accusations here and there that are categorically baseless. You came up with all these manipulation tactics as if you were one of us and knew everything we did. You are also insulting the editors and reviewers as if they did not see this manuscript before they published it, but you Mr genius knew it from the first look. . I hate to talk to someone invisible but I thought I d do it just in case if some reasonable is passing by. Ok let us talk about your unconventional  review . #1: We ran about 10 experiments using B-actin as a loading standard, and we could not run two more!!? This totally insane. Go to the paper and see how many times we ran B-actin. However, despite your welcomed tactics, we have all the raw data and we can send them to the editors if they chose to ask for it #2: Partially similar? Partially !!!! So you mean we only took part of the other figure !?! and why did you make it four bands instead of 5 !?? or maybe it does not fit your narratives. #3: duplication? So you are suggesting superimposing!!? Even your blown up does not fit your story. Look at your magnified picture and compare the bands. How about the next set of bands right beneath it (AKT)!? Do you think we borrowed some bands there too !!? Grow up , It is a new year. It is ok to have some disagreement with someone and not be friendly with that person. However, it is not ok to use this platform for personal goals. Generally speaking, people in academia discuss ideas and may be point out some flaws in a paper but not come out of the gate accusing randomly hoping to smear someone. , 
 We reject all these baseless allegation of figure manipulation Comment #2: We ran about 10 experiments in this paper using B-actin as a loading standard, and we could have ran more if we had too. There was no need to borrow from one figure to the other. The raw data comparison show that this claim is false. (Attachment #1) ![file] Comment#3: Partially similar? Partially !!!! So you mean we only took part of the other figure !?! and why did you make it four bands instead of 5 !?? The attached comparison using the claimed rotation show that this claim is also false. (Attachment #2)![file] Comment#4: duplication? So you are suggesting superimposing!!? Even your blown up figure does not fit your hinted claim. Look at your magnified picture and compare the bands. If you look at the next set of bands right beneath it (AKT) the bands are similar too, but that does not man we used figure manipulation to come up with data!! , 
 #6 Thank you for your reply. Did you also have a response to the concern expressed in #1? , 
 #2 the 2B actin band is different, I think. , 
 #3 https: doi.org 10.3390 ijms22063269, Fig. 3B, HO1 indeed looks strikingly similar to this paper,Fig. 5A, Nuclear Nrf2. I generated an animated overlay to better appreciate the similarity. Click to animate: ![file] , 
 #9 We already debunked this theory. Please see my response above. , 
 Fig3B pPI3K data is made by 180 degrees rotation of GST data in different paper. ![file] , 
 It may be better to see the data carefully.![file] ,