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.
Highlights
Manual acupuncture (MA) has been practiced in China for more than 3000 years and has gained increasing popularity and acceptance in clinical practice worldwide [1, 2]
Our previous studies found that some immune-related cells and cytokines in acupoint changed significantly after MA. e concentrations of TLR4, E-selectin, L-selectin, and cytokines IL-1β, IL-6, IL-8, and TNF-α in the acupoint were significantly increased within a certain time after MA [12, 13], and the number of mast cells and their degranulation rate in the acupoint increased significantly and participated in the MA effect
We have demonstrated that MA has an analgesic, detumescence, and anti-inflammatory effect on CFA-induced inflammatory pain rats and tried to definite the key cells and key cytokines in rat serum responded to MA by complex network and cell-cell communication (CCC) network method
Summary
Manual acupuncture (MA) has been practiced in China for more than 3000 years and has gained increasing popularity and acceptance in clinical practice worldwide [1, 2]. We detected the concentrations of 24 cytokines of ST36 acupoint in CFA-induced inflammatory pain rats after MA treatment by multiplex immunoassay technology and analyzed the data using statistical and complex network and cell-cell communication network diagram method, in order to depict the network relationship between the cytokines and related cells objectively and establish cytokine connection network and CCC network, respectively. On this basis, the key cytokines and key cells of the acupoint at three time-points were selected, and the key cells were detected by flow cytometry and immunofluorescence for localization and quantitative detection. The key cytokines and key cells of the acupoint at three time-points were selected, and the key cells were detected by flow cytometry and immunofluorescence for localization and quantitative detection. is study hopes to provide an accurate and reliable scientific basis for revealing the acupoint initiation mechanism of MA effect
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