Antagonism of cortistatin against cyclosporine-induced apoptosis in rat myocardial cells and its effect on myocardial apoptosis gene expression
Authors: J Yu
X-T Wang
L Geng
Y-L Yang
Journal: European review for medical and pharmacological sciences (2018)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: cortistatin; cyclosporine A; myocardial apoptosis; rats; H9C2 cells
Affiliations: Department of International Medical, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China. .

Abstract:  Objective: To investigate the role of cortistatin (CST) on cyclosporine A (CsA)-induced myocardial apoptosis in rats and determine its effect on the expressions of myocardial apoptosis genes. Materials and methods: H9C2 cells were treated with different concentrations of CsA solution (0.04, 0.2, 1 and 5 μM) for 24, 48 and 72 h, respectively. The cell viability was detected via methyl thiazolyl tetrazolium (MTT) assay, and the appropriate dose and time were compared and determined. At the same time, CST in different concentrations (0.08, 0.04, 0.2, 1, 5 and 25 μM) was added into cell culture, and the appropriate dose was identified using MTT assay. The cellular morphology in each group was observed, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed for the detection of cell apoptosis. Moreover, in molecular mechanism research, the apoptosis-associated factors, factor associated suicide (Fas), Fas ligand (FasL) and B-cell lymphoma-2-associated X protein (Bax), were detected via quantitive Real-time polymerase chain reaction (qPCR). Finally, the levels of a protein related to myocardial apoptosis in rats were investigated via Western blotting. Results: The treatment with 1 μM CsA for 48 h caused significant apoptosis. The results of TUNEL staining showed the inhibitory role of CST on the myocardial apoptosis in rats induced by CsA. The detection of apoptosis factors via Real-time PCR revealed that after the induction of CsA, the expressions of Fas, FasL and Bax mRNA in cells were significantly higher than those in control group, but were significantly decreased after administration of CST. Western blotting showed that the protein expressions of Caspase 3 and Caspase 9 were remarkably elevated in cells after the use of CsA, but were significantly reduced after administration of CST (p &lt; 0.01). Conclusions: CST contributes to antagonistic function against the CsA-induced apoptosis of rat myocardial cells, and its effect is related to the down-regulation of expressions of apoptotic factors, Fas, FasL, Bax, Caspase 3, and Caspase 9. 

Retracted: No

PubPeer comments:
 In Figure 3, areas in micrographs have been duplicated, manipulated and presented as something else. In Figure 4, an area of a flow cytometry dot plot has been duplicated and presented as something else. In Figure 10, a Western immunoblot panel has been duplicated and presented as something else (AKT VS ACTIN) ![file] , 
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 *[right]* Figure 3 again.  AO EB dual staining assay for determination of cellular morphology and apoptosis after Cinnamolide treatment to human cervical HeLa cancer cells. The results indicate membrane blebbing, increased number of yellowgreen (early apoptotic cells), orange (late apoptotic cells) and orange-red (necrotic cells) fluorescent cells. Arrows indicate apoptotic cells. I have repeated two panels awith 180-deg flips. Colored rectangles added to show unexpected similarities with... *[left]* Figure 6 from [Tan et al (2019)] .  Wogonin triggers apoptosis in SW48 colorectal cancer cells as depicted by AO EB staining. ... Green arrow indicates early apoptotic cells; blue arrow shows late apoptotic cells; purple arrow shows membrane blebbing; and red arrow shows loss of membrane shape. ![file] , 
 *[right]* Figure 3 again. One panel repeated with a 180-deg flip. Colored rectangles added to show unexpected similarity with... *[left]* Figure 3 of [Tian et al (2019)] .  Scopoletin induces apoptotic cell death in HeLa cervical cancer cells as depicted by AO EB staining. Green arrows indicate early apoptotic cells, blue arrows depict late apoptotic cells, purple arrows depict membrane blebbing, yellow arrows depict cells which appear shrunken and red arrows depict loss of membrane shape. ![file] , 
 *[left]* Fig 4 again. Though extensively re-worked, the panels appear to be based on ... *[right]* Fig 4 from [Zhang et al (2018)] .  Estimation of apoptotic cell population by annexin V PI staining at indicated doses of lactucopicrin. ... The results indicate that with increasing doses of the compound, the population of both early and late apoptotic cells increased significantly . ![file] UPDATE: Another sighting! Fig 5 from [Wang et al (2018)] .  Estimation of apoptotic cell populations by annexin V PI staining of HCT-116 cells using flow cytometry...The results revealed that the apoptotic cell populations increased from 3.04% in the controls to 33.9% at 30u03bcM concentration. ![file] , 
 #3 Another sighting! Fig 1D of [Wu et al (2020)] . ![] , 
 *[left]* Figure 2. A:  Clonogenic assay for determination of anticolony potential of Cinnamolide in human cervical HeLa cancer cells. Cinnamolide treatment led to a significant and dose-dependent reduction in the number of cancer cell colonies . *[right]* Fig 2A of [Li et al (2020)] .  miR-7 induces apoptosis in TPC-1 cancer cells. A. Colony formation by TPC-1 cells transfected with miRNC and miR-7 mimics . Plus 180-deg-flipped version. ![file] Two panels are repeated. UPDATED: Another sighting! Fig 3E of [Liu, Liu Zhao (2020)] .  Colony formation of the NC or Si-MAPK1 transfected DoTc2 cells . ![] , 
 The paper has undergone a stealth revision or correction, replacing Figures 2, 3 and 4, and the AKT band of Fig 10. The version currently provided on the journal website was created modified on 22 January 2021. ![file] At the time of writing, the original Figures are still cached in Google Image Search, though this will change. ![file] , 
 The paper has reverted to its original version, and acquired a [May June Expression of Concern] .  Following the publication of this article [1], readers drew to our attention that part of the data was possibly unreliable. We sent emails to the authors with a request to provide the raw data to prove the originality, but received no reply. Therefore, as we continue to work through the issues raised, we advise readers to interpret the information presented in the article with due caution. We thank the readers for bringing this matter to our attention. We apologize for any inconvenience it may cause. ,