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Protection of pancreatic β-cell by phosphocreatine through mitochondrial improvement via the regulation of dual AKT/IRS-1/GSK-3β and STAT3/Cyp-D signaling pathways

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Abstract

Diabetes mellitus (DM) is a metabolic syndrome, caused by insufficient insulin secretion or insulin resistance (IR). DM enhances oxidative stress and induces mitochondrial function in different kinds of cell types, including pancreatic β-cells. Our previous study has showed phosphocreatine (PCr) can advance the mitochondrial function through enhancing the oxidative phosphorylation and electron transport ability in mitochondria damaged by methylglyoxal (MG). Our aim was to explore the potential role of PCr as a molecule to protect mitochondria from diabetes-induced pancreatic β-cell injury with insulin secretion deficiency or IR through dual AKT/IRS-1/GSK-3β and STAT3/Cyclophilin D (Cyp-D) signaling pathways. MG-induced INS-1 cell viability, apoptosis, mitochondrial division and fusion, the morphology, and function of mitochondria were suppressed. Flow cytometry was used to detect the production of intracellular reactive oxygen species (ROS) and the changes of intracellular calcium, and the respiratory function was measured by oxygraph-2k. The expressions of AKT, IRS-1, GSK-3β, STAT3, and Cyp-D were detected using Western blot. The result showed that the oxidative stress–related kinases were significantly restored to the normal level after the pretreatment with PCr. Moreover, PCr pretreatment significantly inhibited cell apoptosis, decreased intracellular calcium, and ROS production, and inhibited mitochondrial division and fusion, and increased ATP synthesis damaged by MG in INS-1 cells. In addition, pretreatment with PCr suppressed Cytochrome C, p-STAT3, and Cyp-D expressions, while increased p-AKT, p-IRS-1, p-GSK-3β, caspase-3, and caspase-9 expressions. In conclusion, PCr has protective effect on INS-1 cells in vitro and in vivo, relying on AKT mediated STAT3/ Cyp-D pathway to inhibit oxidative stress and restore mitochondrial function, signifying that PCr might become an emerging candidate for the cure of diabetic pancreatic cancer β-cell damage.

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The data and material used and/or analyzed during the current study are available from the corresponding authors on reasonable request.

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Funding

This work was financially supported by the Natural Science Foundation of China (30772601) and the University Innovation Team Project Foundation of Education Department of Liaoning Province (LT2013019).

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  1. Hongyan Wang, Jie Ai and Abdullah Shopit contributed equally to this work

Authors and Affiliations

  1. Acad Integrated Med & College of Pharmacy, Department of Pharmacology, Dalian Medical University, 9 Western Section, Lvshun South Street, Dalian, 116044, China

    Hongyan Wang, Jie Ai, Abdullah Shopit, Mengyue Niu, Nisar Ahmed, Tsehaye Tesfaldet, Yazeed Jamalat, Peng Chu, Jinyong Peng, Xiaodong Ma, Eskandar Qaed, Guozhu Han & Zeyao Tang

  2. Department of Jilin University, Changchun, China

    Zhongyuan Tang

  3. Second Clinical College, Dalian Medical University, Dalian, China

    Xiaodong Li

  4. First Clinical College, Dalian Medical University, 9 Western Section, Lvshun South Street, Dalian, 116044, China

    Weisheng Zhang

  5. Department of Pathophysiology, Dalian Medical University, 9 Western Section, Lvshun South Street, Dalian, 116044, China

    Jun Wang

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  1. Hongyan Wang
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Contributions

H.W., J.A., and A.S. share first authorship. H.W., J.A., and A.S. designed and directed the experimental studies. H.W. and J.A., performed in vitro cell line-based studies. H.W., J.A., A.S., M.N., Z.T., and X.L. performed the genes expression studies. H.W., A.S., N.A., Y.J., and P.C. performed statistical analysis and interpreted the data. H.W., J.A., A.S., Z.T., X.L., H.L., and J.P. executed the in-vivo experiments. X.M., T.T., and E.A. performed the immunofluorescence and confocal experiments. H.W., J.A., A.S., M.N., and T.T. wrote the manuscript. Z.T., J.W., W.Z., and G.H. directed the overall project. Z.T., J.W., and W.Z. author corresponding.

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Correspondence to Weisheng Zhang, Jun Wang or Zeyao Tang.

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Highlights

• PCr possesses a protective effect on the damage of INS-1 cells in vitro and in vivo.

• PCr against apoptosis and oxidative stress also improves mitochondrial function.

• The AKT/IRS-1/GSK-3β and STAT3/Cyp-D signaling pathways play a central role in the protection of PCr.

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Wang, H., Ai, J., Shopit, A. et al. Protection of pancreatic β-cell by phosphocreatine through mitochondrial improvement via the regulation of dual AKT/IRS-1/GSK-3β and STAT3/Cyp-D signaling pathways. Cell Biol Toxicol 38, 531–551 (2022). https://doi.org/10.1007/s10565-021-09644-7

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