Stem Cell Reviews and Reports

, Volume 14, Issue 4, pp 558–573 | Cite as

The Delta Opioid Peptide DADLE Represses Hypoxia-Reperfusion Mimicked Stress Mediated Apoptotic Cell Death in Human Mesenchymal Stem Cells in Part by Downregulating the Unfolded Protein Response and ROS along with Enhanced Anti-Inflammatory Effect

  • Madhubanti Mullick
  • Dwaipayan Sen


Hypoxia-reperfusion (H/R) emblems a plethora of pathological conditions which is potent in contributing to the adversities encountered by human mesenchymal stem cells (hMSCs) in post-transplant microenvironment, resulting in transplant failure. D-Alanine 2, Leucine 5 Enkephaline (DADLE)-mediated delta opioid receptor (DOR) activation is well-known for its recuperative properties in different cell types like neuronal and cardiomyocytes. In the current study its effectiveness in assuaging hMSC mortality under H/R-like insult has been delineated. The CoCl2 mimicked H/R conditions in vitro was investigated upon DOR activation, mediated via DADLE. hMSCs loss of viability, reactive oxygen species (ROS) production, inflammatory responses and disconcerted unfolded protein response (UPR) were assessed using AnnexinV/PI flow cytometry, fluorescence imaging, mitochondrial complex 1 assay, quantitative PCR, immunoblot analysis and ELISA. H/R like stress induced apoptosis of hMSCs was significantly mitigated by DADLE via modulation of the apoptotic regulators (Bcl-2/Bax) along with significant curtailment of ROS and mitochondrial complex 1 activity. DADLE concomitantly repressed the misfolded protein aggregation, alongside the major UPR sensors: PERK/BiP/IRE-1α /ATF-6, evoked due to the H/R mimicked endoplasmic reticulum stress. Undermined phosphorylation of the Akt signalling pathway was observed, which concerted its effect onto regulating both the pro and anti-inflammatory cytokines, actuated as a response to the H/R-like insult. The effects of DADLE were subdued by naltrindole (specific DOR antagonist) reaffirming the involvement of DOR in the process. Taken together these results promulgate the role of DADLE-induced DOR activation on improved hMSC survival, which signifies the plausible implications of DOR-activation in cell-transplantation therapies and tissue engineering aspect.


DADLE Delta opioid peptide hMSCs Hypoxia-reperfusion Survival Transplantation 



human mesenchymal stem cells




d-Ala2, d-Leu5 enkephalin


Delta opioid receptor


Endoplasmic reticulum


Unfolded protein response


chloro-methyl derivative 2′,7′-dichlorofluorescein


PRKR-like ER kinase


Activating transcription factor 6α


Inositol-requiring protein 1α


immunoglobulin heavy-chain binding protein


Mitogen activated protein kinase



Centre for Stem Cell Research, Christian Medical College, Vellore, India for providing access to flow cytometry and fluorescence microscope.

Authors’ contribution

MM has carried out experiments, analyzed data and wrote the paper. DS conceptualized, wrote the paper and analysed the data.


This work was supported by a start-up fund from VIT, Vellore given to DS. DS is also supported by a Indian Council of Medical Research (ICMR) Funded Project (Sanction Order No.NCD/Ad-hoc/66/2016–17) and a ‘Fast Track Young Scientist’ grant (YSS/2014/000027) from the Science and Engineering Research Board (SERB), Department of Science and Technology (DST), Government of India.

Compliance with Ethical Standards

Competing Interests

The authors declare that there are no conflicts of interest.

Consent for Publication

All authors have read the manuscript and agreed to the publication.


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Authors and Affiliations

  1. 1.Cellular and Molecular Therapeutics Laboratory, Centre for Biomaterials Cellular and Molecular Theranostics (CBCMT)Vellore Institute of Technology (VIT)VelloreIndia

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