Applying Complex Network and Cell‐Cell Communication Network Diagram Methods to Explore the Key Cytokines and Immune Cells in Local Acupoint Involved in Acupuncture Treating Inflammatory Pain

Abstract

Manual acupuncture (MA) can effectively treat a variety of diseases, but its specific mechanism remains unclear. The “acupoint network” activated by MA participates in MA signal transduction, in which immune-related cells and cytokines play an important role. However, which cells and cytokines in the acupoint have changed after MA? What is the network relationship between them? Which cells and cytokines may play the most important role in MA effect? These problems are unclear. In this study, on the basis of affirming the analgesic, detumescence, and anti-inflammatory effect of MA, the concentration of 24 cytokines in ST36 acupoint in rats with inflammatory pain after MA treatment was detected by multiplex immunoassay technology. Then, using statistical and complex network and cell-cell communication (CCC) network diagram method to analyze the detected data depicts the network relationship between the cytokines and related cells objectively and establishes cytokine connection network and CCC network, respectively. The results showed that MA reinforced communication intensity between cells while reducing the overall correlation intensity. On this basis, the key cytokines and key cells at three MA time-points were screened out, cytokines IL-6, MCP-1, fibroblasts cell, and monocyte macrophage screened by the three methods at three MA time-points might be the key cytokines or key cells. After that, we detected the macrophages in ST36 acupoint by flow cytometry and immunofluorescence and found that the relative amount of macrophages increased significantly after MA, especially the macrophage of the dermis of skin. This study provided a basis for revealing the initiated mechanism of MA effect.

The Anti-Inflammatory Actions and Mechanisms of Acupuncture from Acupoint to Target Organs via Neuro-Immune Regulation

“…Fibroblasts and macrophages are the key cells that are stimulated with MA stimulation of the ST36 acupoint. 114 Gene chip assays revealed that the MA stimulation of ST36 modulates the expression of 236 genes and seven signaling pathways (ERK, P38, JNK, P53, BCR, TCR, and TLR receptor signaling), among which the ERK pathway was the most notable. 115 Apart from this, the upregulation of NF-κB signaling and the related cytokine gene expression (p-p65, p-p50, Nikbia, IL-1β, IL-6, CXCL1, CCL2, Rela, and NF-κB1) are also associated with ST36 stimulation.…”

Anti-Inflammatory Effects of Acupuncture at ST36 Point: A Literature Review in Animal Studies

“…MA at ST36 increased MCP-1, CXCL1, MIP-3α, MIP-1α, RANTES, IL-1β, IL-1α, IL-6, IL-2, IL-12, IL-13, granulocyte colony-stimulating factor (G-CSF), and M-CSF and decreased IL-18, IL-10, CRP, IFN-γ, GM-CSF, and percentage of CD45+CD11b+CD68+ cells in ST36 acupoint tissues. This study revealed the key changes of immune-related cells, cytokines, chemokines, and macrophages caused by MA and defined that MA effects are mediated by regulating cell–cell communication (CCC) networks ( 87 ). In another CFA-injected inflammatory pain model, ST36 MA increased TRPV1, TRPV4, ASIC3, PanX1, Cx43, and P2Y1 and decreased PGP 9.5 in muscle tissues at ST36.…”

The immunomodulatory mechanisms for acupuncture practice

“…Zhang et al. treated the complete Freund’s Adjuvant (CFA)-induced inflammatory pain rats with manual acupuncture (MA), and they found that the inflammatory pain was significantly relieved, and the amounts of macrophages at acupoints were also raised in the experimental group ( 23 ). Interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1, and IL-1 are important cytokines that have been identified by statistical and cytokine association networks.…”