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Ropivacaine Induces Cell Cycle Arrest in the G0/G1 Phase and Apoptosis of PC12 Cells via Inhibiting Mitochondrial STAT3 Translocation

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Abstract—

STAT3 has neuroprotective effect via non-canonical activation and mitochondrial translocation, but its effect on ropivacaine-induced neurotoxicity remains unclear. Our previous study revealed that apoptosis was an important mechanism of ropivacaine-induced neurotoxicity; this study is to illustrate the relationship between STAT3 with ropivacaine-induced apoptosis. Those results showed that ropivacaine treatment decreased cell viability, induced cell cycle arrest in the G0/G1 phase, apoptosis, oxidative stress, and mitochondrial dysfunction in PC12 cells. Moreover, ropivacaine decreased the phosphorylated levels of STAT3 at Ser727 and downregulated the expression of STAT3 upstream gene IL-6. The mitochondrial translocation of STAT3 was also hindered by ropivacaine. To further illustrate the connection of STAT3 protein structure with ropivacaine, the autodock-vina was used to examine the interaction between STAT3 and ropivacaine, and the results showed that ropivacaine could bind to STAT3’s proline site and other sites. In addition, the activator and inhibitor of mitoSTAT3 translocation were used to demonstrate it was involved in ropivacaine-induced apoptosis; the results showed that enhancing the mitochondrial STAT3 translocation could prevent ropivacaine-induced apoptosis. Finally, the expression of p-STAT3 and the levels of apoptosis in the spinal cord were also detected; the results were consistent with the cell experiment; ropivacaine decreased the expression of p-STAT3 protein and increased the levels of apoptosis in the spinal cord. We demonstrated that ropivacaine induced apoptosis by inhibiting the phosphorylation of STAT3 at Ser727 and the mitochondrial STAT3 translocation. This effect was reversed by the activation of the mitochondrial STAT3 translocation.

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Data Availability

Data that support the study findings are available from the corresponding author upon reasonable request.

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Funding

The Innovative Research Program for Graduates of Hubei University of medicine (NO. YC2020029), Hubei Natural Science Foundation (No. 2019CFB105).

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Author notes

  1. Lian Zeng, Aohan Li, and Zhen Zhang contributed equally to the work and should be regarded as co-first authors.

Authors and Affiliations

  1. Department of Anesthesiology, Xiangyang Key Laboratory of Movement Disorders, Xiangyang No.1 People’s Hospital, Hubei Clinical Research Center of Parkinson’s Disease, Hubei University of Medicine, Hubei, China

    Lian Zeng, Aohan Li, Zhen Zhang, Fuyu Zhang, Ying Wang & Huiyu Luo

  2. Department of Oncology, Xiangyang No.1 People’s Hospital, Hubei University of Medicine, Hubei, China

    Huaxian Chen & Xudong Ding

  3. Department of Rehabilitation Medicine, Xiangyang No.1 People’s Hospital, Hubei University of Medicine, Hubei, China

    Huiyu Luo

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Contributions

LHY and DXD designed the studies; ZL, LAH, and ZZ undertook the cell experiments and the construction of animal experiment; ZL and ZFY undertook the molecular biology testing; ZL, CHX, and WY undertook the molecular docking. ZL, ZFY, and ZZ analyzed data and wrote the draft of manuscript; ZL and LAH undertook the revision of manuscript.

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Correspondence to Huiyu Luo.

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All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at the Animal Ethics Committee of the Hubei University of Medicine (permit number 2018DW003).

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10753_2021_1508_MOESM1_ESM.pdf

Supplementary file1 Supplementary Fig. 1 Ropivacaine promotes the protein expression of Cytochrome c and cleaved-PARP. A: The protein expression of Cytochrome c, PARP and GAPDH was detected by Western-blot. B: Quantification of the gray values in graphs A. C: Effect of ropivacaine on the colocalization of mitochondria (red) and Cytochrome c (green) was detected by fluorescence microscope. (PDF 8043 KB)

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Zeng, L., Li, A., Zhang, Z. et al. Ropivacaine Induces Cell Cycle Arrest in the G0/G1 Phase and Apoptosis of PC12 Cells via Inhibiting Mitochondrial STAT3 Translocation. Inflammation 44, 2362–2376 (2021). https://doi.org/10.1007/s10753-021-01508-w

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