Abstract
This study aims to explore the regulatory mechanisms of dexmedetomidine in parthanatos. MTT assay was applied to reveal cell viability; JC-1 staining assay was utilized to reveal mitochondrial membrane potential. Reactive oxygen species (ROS) probe, DCFH-DA, was used to detect intracellular ROS production. Luciferase activity assay was applied to measure the binding between miR-7-5p and PARP1. We first identified that bupivacaine inhibited the viability and induced the parthanatos of human neuroblastoma SH-SY5Y cells. In addition, dexmedetomidine, a potent α2-adrenoceptor agonist, reversed the regulatory effect of bupivacaine on parthanatos of SH-SY5Y. More importantly, dexmedetomidine counteracted bupivacaine-induced changes of mitochondrial membrane potential and ROS production in SH-SY5Y cells. Hyper-activation of PARP1 plays a vital role in parthanatos. Further exploration of our study identified that bupivacaine triggered overexpression of PARP1 in SH-SY5Y cells. Bioinformatics analysis revealed that miR-7-5p targeted the 3′ untranslated region (3′ UTR) of PARP1 to inhibit PARP1 expression. In addition, dexmedetomidine recovered the suppressive effects of bupivacaine on miR-7-5p expression. Dexmedetomidine suppressed bupivacaine-induced parthanatos in SH-SY5Y cells via the miR-7-5p/PARP1 axis, which may shed a new insight into parthanatos-dependent neuronal injury.
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01 April 2021
A Correction to this paper has been published: https://doi.org/10.1007/s00210-021-02080-8
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Acknowledgments
We appreciate the support of Fujian Medical University, Fujian Provincial Hospital and Fujian Provincial Emergency Center.
Funding
This work was supported by the
1. The Medical Education Branch of the Chinese Medical Association, the Medical Education Committee of the Chinese Higher Education Society (2016B-LC033)
2. the Natural Science Foundation of Fujian Province (2018J01246)
3. the Medical Innovation Project of Fujian province (2018-CX-2)
4. high-level hospital foster grants from Fujian Provincial Hospital, Fujian Province, China (2019HSJJ23)
5.high-level hospital foster grants from Fujian Provincial Hospital, Fujian Province, China (2019HSJJ21)
6. the Training Project for Talents of Fujian Provincial Health Commission (2019-ZQN-1)
7.Provincial special subsidy funds for health care of Fujian Provincial Department of Finance (No. 2020-467)
8. Fujian Science and Technology Innovation Joint Fundation (Major Program)(2019Y9028)
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Study concepts and design: Ting Zheng and Xiao-chun Zheng
Data acquisition and analysis: Ting Zheng, Chunying Zheng, and Fei Gao
Experimental studies and statistical analysis: Ting Zheng, Chunying Zheng, Fei Gao, and Fengyi Huang
Manuscript preparation and editing: Ting Zheng, Chunying Zheng, Fei Gao, Fengyi Huang, and Bin Hu
Manuscript review: Ting Zheng, Chunying Zheng, Fei Gao, Fengyi Huang, Bin Hu, and Xiaochun Zheng
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Supplementary Figure 1
(A) Cell viability by treatment of Bupivacaine for different time was revealed. N = 5. (B) RT-qPCR revealed expression of MIR7-3HG by Bupivacaine and (or) Dexmedetomidine treatment. #p < 0.05 compared with DMSO group; Фp < 0.05 compared with Bupivacaine group or Dexmedetomidine group. N = 5. (C) RT-qPCR revealed expression of EZH2 by Bupivacaine and (or) Dexmedetomidine treatment. #p < 0.05 compared with DMSO group; Фp < 0.05 compared with Bupivacaine group or Dexmedetomidine group. N = 5. (D) RT-qPCR revealed expression of EZH2 by transfection of EZH2#1/2 in Bupivacaine (10 μM) treated SH-SY5Y cells. #p < 0.05 compared with sh-NC group. (E-F) RT-qPCR revealed the influence of EZH2#1/2 on expression of MIR7-3HG and miR-7-5p in Bupivacaine (10 μM) treated SH-SY5Y cells. #p < 0.05 compared with sh-NC group. (G) Luciferase reporter assay revealed activity of MIR7-3HG promoter under the influence of EZH2#1/2 in Bupivacaine (10 μM) treated cells. #p < 0.05 compared with sh-NC group. (H) ChIP assay revealed relative enrichment of MIR7-3HG promoter precipitated by IgG, EZH2 and H3K27me3. #p < 0.05 compared with IgG group. (PNG 459 kb)
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Zheng, T., Zheng, C., Gao, F. et al. Dexmedetomidine suppresses bupivacaine-induced parthanatos in human SH-SY5Y cells via the miR-7-5p/PARP1 axis-mediated ROS. Naunyn-Schmiedeberg's Arch Pharmacol 394, 783–796 (2021). https://doi.org/10.1007/s00210-020-01971-6
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DOI: https://doi.org/10.1007/s00210-020-01971-6