Regulatory effects and mechanisms of the acupuncture-induced complement component 1q on the high mobility group box 1 -receptor for advanced glycation end products-nuclear factor kappa-B pathway at Zusanli (ST36).

چکیده اصلی

OBJECTIVE: To investigate what are the crucial regulators and mechanisms involved in regulating the inflammatory responses at Zusanli (ST36). METHODS: Initially, we utilized enzyme-linked immunosorbent assay (ELISA) to detect the variations of high mobility group box 1 (HMGB1), interleukin-10 (IL-10), and complement component 1q (C1q) concentrations at Zusanli (ST36) on day 1 (D1), day 3 (D3), day 5 (D5) and day 7 (D7) after acupuncture to clearly depict the time-dependent changes of these inflammatory substances. Subsequently, we utilized C1qa-knockout (C1qa-KO) mice to investigate the role of acupuncture-induced C1q in inflammatory responses at Zusanli (ST36). We utilized hematoxylin and eosin staining to observe the inflammatory morphological alterations, examined the activation of the NF-κB pathway by immunofluorescence and assessed HMGB1 and IL-10 changes using ELISA. Lastly, to uncover the specific mechanism by which C1q regulates the acupuncture-induced inflammatory responses at Zusanli (ST36), we employed Western blot (WB) to measure the expression levels of receptor of advanced glycation end products (RAGE), leukocyte associated immunoglobulin like receptor 1 (LAIR-1), and src homology region 2 domain-containing protein tyrosine phosphatase 1 (SHP-1) proteins and used immunofluorescence to detect the polarization of classically activated macrophages (M1) and alternatively activated macrophages (M2). RESULTS: Compared with the wild-type group, the levels of HMGB1 and C1q of acupuncture (ACU) group at Zusanli (ST36) were increased on D3 after acupuncture, while the level of IL-10 decreased. With continued higher level of HMGB1, both C1q and IL-10 level were increased on D7. After knocking out C1q, compared with the ACU group, the C1qa-KO+acupuncture (C1q-/-+ACU) group exhibited further aggravated inflammatory infiltrations in the epidermal layer, dermal layer, and muscular layer. At Zusanli (ST36), the level of HMGB1 was further increased, accompanied by an intensified activation of the NF-κB pathway. Meanwhile, the level of IL-10 exhibited a downward tendency, which jointly led to the exacerbation of inflammatory responses at Zusanli (ST36). In comparison with the ACU group, the C1q-/-+ACU group showed a reduction in SHP-1 levels, a downward tendency of LAIR-1 expression, whereas the expression of RAGE exhibited an upward inclination. Furthermore, the number of M1 macrophages at Zusanli (ST36) increased significantly, in stark contrast to a pronounced decline in the number of M2 macrophages. CONCLUSION: Zusanli (ST36) acupuncture initiates a time-regulated inflammatory cascade might through the following mechanism: HMGB1 release (D1) triggers C1q upregulation (D3), culminating on D7 with C1q binding to LAIR-1, which recruits SHP-1 to inhibit HMGB1-RAGE-NF-κB signaling viaM2 polarization and IL-10 feedback, ensuring precisely controlled inflammation at acupoints.

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