Modified Danggui Sini formula promotes wound healing regulating natural killer cell function: a network pharmacology-based study targeting killer cell lectin like receptor K1
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Published on: February 22, 2020
X U Yannan1,2,3, Tian Yuan1,2, Wang Xionghui1,2,4
1Department of Chinese Medicine, Changhai Hospital, Naval Medical University, Shanghai 200433, China.
Objective:
To explore the potential mechanism of the modified Danggui Sini formula (, M-DGSNF) in the treatment of freezing cold injury (FCI).
Methods:
FCI model rats were used to evaluate the effects of M-DGSNF on wound appearance viahistopathological analysis. Furthermore, the potential targets of M-DGSNF were identified viaultra-performance liquid chromatography quadrupole-time-flight mass spectrometry (UPLC-Q-TOF/MS) analysis, network pharmacology and RNA sequencing. Frozen mouse skin sections were analysed by Western blotting and immunohistochemistry. Interleukin (IL)-6, interferon-gamma (IFN-γ), IL-4, IL-10 and vascular endothelial growth factor (VEGF) were assessed by enzyme-linked immunosorbent assay. The effects of M-DGSNF on macrophage polarization were monitored in vitroby flow cytometry. Depletion of natural killer (NK) cells in vivo was used to confirm the role of NK cells in the effects of M-DGSNF.
Results:
M-DGSNF promoted wound healing and ameliorated the inflammatory response to frostbite wounds in rats with FCI. Additionally, the expression of CD31, alpha-smooth muscle actin (α-SMA), and type I collagen, which promote wound healing, increased after M-DGSNF treatment. Furthermore, killer cell lectin-like receptor k1 (KLRK1) and indoleamine 2,3-dioxygenase 1 (IDO1), which are associated with the function of NK cells, were identified by UPLC-high-resolution mass spectro-metry analysis, network pharmacology and RNA-seq. Accordingly, an increase in the release of inflammatory factors such as VEGF, IFN-γ, IL-6, IL-4, and IL-10 was observed in the early stage of FCI, after which the expression of IFN-γ and IL-6 decreased in the late stage of frostbite. Furthermore, M-DGSNF treatment contributed to M2 macrophage polarization both in vivo and in vitro. Depletion of NK cells abolished the effects of M-DGSNF on wound healing, suggesting that NK cells play a key role in the mechanism by which M-DGSNF affects mice with FCI.
Conclusion:
M-DGSNF may promote wound healing in mouse models of FCI by regulating the function of NK cells through targeting KLRK1.
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