Analysis of the circRNA/lncRNA–miRNA–mRNA Networks of Action of Fire Needle Acupuncture in Relieving Chemotherapy-Induced Neuropathic Pain

Mengyu Yang,1,* Yuexuan Chen,2,* Jingjing Li,3 Ruilin Zhang,4– 6 Xindong Wang,7 Jieshan Guan,5,6,8 Zhijie Luo,5,6,8 Jingchun Zeng4– 6 1Clinical Research and Big Data Laboratory, South China Research Center for Acupuncture and Moxibustion, Medical College of Acu-Moxi and Rehabilitation, Guangzhou University of Chinese Medicine, Guangzhou, 510006, People’s Republic of China; 2Department of Acupuncture and Moxibustion, Shenzhen Hospital (Futian) of Guangzhou University of Chinese Medicine, Shenzhen, 518000, People’s Republic of China; 3Bao’an Traditional Chinese Medicine Hospital, Seventh Clinical Medical College of Guangzhou University of Traditional Chinese Medicine, Shenzhen, 518133, People’s Republic of China; 4Department of Rehabilitation, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510405, People’s Republic of China; 5Guangdong Clinical Research Academy of Chinese Medicine, Guangzhou, 510405, People’s Republic of China; 6First Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou, 510405, People’s Republic of China; 7Nanyang Institute of Technology, Zhang Zhongjing College of Chinese Medicine, Acupuncture and Massage Teaching and Research of Office, Nanyang, 473004, People’s Republic of China; 8Department of Oncology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510405, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jingchun Zeng, Department of Rehabilitation, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, No. 16, Airport Road, Baiyun District, Guangzhou, Guangdong, 510405, People’s Republic of China, Email [email protected]: The current treatment strategies of chemotherapy-induced peripheral neuropathy (CIPN) are not ideal. Fire needle acupuncture (FA) has been shown to have significant therapeutic effects on pain, including cancer-related pain. However, the potential mechanisms by which FA alleviates CIPN remain unclear.Methods: A CIPN model was established using oxaliplatin, followed by FA treatment. Mechanical sensitivity in rats was assessed using the paw withdrawal threshold (PWT), and inflammatory cell infiltration in the L4–L6 spinal cord segments was analyzed via hematoxylin and eosin staining. Subsequently, L4–L6 spinal cord segments were collected and whole-transcriptome sequencing and small RNA sequencing were performed to identify differentially expressed genes. PPI networks, circRNAs/lncRNAs–miRNAs–mRNAs, and functional analyses of hub genes were performed to elucidate the potential mechanisms through which FA therapy improves CIPN.Results: FA therapy rapidly increased the PWT and reduced inflammatory infiltration of the spinal cord in CIPN rats. Additionally, 656, 192, 15, and 657 differentially expressed circRNAs, lncRNAs, miRNAs, and mRNAs, respectively, were identified as being associated with FA treatment of CIPN. FA treatment is associated with extracellular matrix receptor interactions, the PI3K–Akt and IL-17 pathways. Furthermore, T cells, macrophages, and T follicular helper cells were closely associated with hub genes, suggesting that these immune cells play a pivotal role in the FA-mediated regulation of CIPN.Conclusion: This study elucidates the potential mechanisms by which FA therapy alleviates CIPN. This study provides a direction for future research on FA therapy and lays a theoretical foundation for the clinical application of FA. This leads to data analysis involving whole-transcriptome sequencing and smallRNA sequencing. The analysis includes GO and KEGG analysis, identifying DE-genes, DE-circRNAs, DE-miRNAs and DE-lncRNAs. These elements contribute to constructing a PPI network and CircRNAs/lncRNAs–miRNAs–mRNAs regulatory networks. The process identifies a hub gene, which is linked to immune infiltration. Finally, differential gene expression validation is performed using spinal cord samples analyzed by qRT-PCR.Flowchart of chemotherapy-induced neuropathy treatment and gene expression analysis in rats.Keywords: chemotherapy, peripheral neuropathy, acupuncture, immune, inflammation