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Heterogeneity of cannabinoid ligand-induced modulations in intracellular Ca2+ signals of mouse pancreatic acinar cells in vitro

Acta Pharmacologica Sinica (2018) | Download Citation

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Abstract

We recently reported that a CB2R agonist, GW405833 (GW), reduced both the ACh-induced Ca2+ oscillations and the L-arginine-induced Ca2+ signal enhancement in mouse pancreatic acinar cells, suggesting that GW-induced inhibition may prevent the pathogenesis of acute pancreatitis. In this study, we aim to evaluate the effects of other cannabinoid ligands on Ca2+ signaling in acinar cells. Patch-clamp whole-cell recordings were applied to measure ACh-induced intracellular Ca2+ oscillations in pancreatic acinar cells acutely dissociated from wild-type (WT), CB1R knockout (KO), and CB2R KO mice, and the pharmacological effects of various cannabinoid ligands on the Ca2+ oscillations were examined. We found that all the 8 CB2R agonists tested inhibited ACh-induced Ca2+ oscillations. Among them, GW, JWH133, and GP1a caused potent inhibition with IC50 values of 5.0, 6.7, and 1.2 μmol/L, respectively. In CB2R KO mice or in the presence of a CB2R antagonist (AM630), the inhibitory effects of these 3 CB2R agonists were abolished, suggesting that they acted through the CB2Rs. The CB1R agonist ACEA also induced inhibition of Ca2+ oscillations that existed in CB1R KO mice and in the presence of a CB1R antagonist (AM251), suggesting a non-CB1R effect. In WT, CB1R KO, and CB2R KO mice, a nonselective CBR agonist, WIN55,212-2, inhibited Ca2+ oscillations, which was not mediated by CB1Rs or CB2Rs. The endogenous cannabinoid substance, 2-arachidonoylglycerol (2-AG), did not show an inhibitory effect on Ca2+ oscillations. In conclusion, CB2R agonists play critical roles in modulating Ca2+ signals in mouse pancreatic acinar cells, while other cannabinoid ligands modulate Ca2+ oscillations in a heterogeneous manner through a CB receptor or non-CB-receptor mechanism.

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Acknowledgements

This work was partially supported by the Shantou University Seed Fund. The authors thank Karen Vu for his assistance in editing the English.

Author contributions

K.-k.X. performed patch-clamp recording, data analysis, made figures, and wrote initial manuscript; J.-x.S. performed some experiments, data analysis, and wrote part manuscript; Z.-b.H. performed some experiments and analysis data; H.-m.S. performed experiments and data analysis; M.G. performed experiments and data analysis; D.-j.C. performed data analysis; S.-j.Z. designed experiments and revised manuscript; J.W. designed experiments, data analysis, made and revised figures, and wrote and revised the manuscript.

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Author notes

  1. These authors contributed equally: Kun-kun Xia, Jian-xin Shen

Affiliations

  1. Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

    • Kun-kun Xia
    • , Shui-jun Zhang
    •  & Jie Wu
  2. Department of Neurobiology, Barrow Neurological Institute, St. Joseph’s Hospital and Medical Center, Phoenix, AZ, 8501, USA

    • Kun-kun Xia
    • , Ze-bing Huang
    • , Ming Gao
    • , De-jie Chen
    •  & Jie Wu
  3. Department of Physiology, Shantou University Medical College, Shantou, 515100, China

    • Jian-xin Shen
    • , Hui-min Song
    •  & Jie Wu
  4. Department of Neurology, Yunfu People’s Hospital, Yunfu, 527300, China

    • De-jie Chen
    •  & Jie Wu

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The authors declare no competing interests.

Corresponding author

Correspondence to Jie Wu.

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DOI

https://doi.org/10.1038/s41401-018-0074-y