Network pharmacology and animal experiments revealed the protective effects of Guilong prescription on chronic prostatitis and its possible mechanisms.

Abstract

Objective: To investigate the protective effects of Guilong prescription (, GL) on chronic prostatitis (CP) and unravel the underlying mechanisms of its pharmacological effects.

Methods: The composition of GL was determined via linear ion trap/electrostatic field orbital trap tandem high-resolution mass spectrometry, and the identified compounds were performed network pharmacological analysis to predict possible pathways of the effects of GL on CP. A CP rat model was established by carrageenan, and rats were randomly assigned into a Control group, Sham group, CP group, GL low dose (3.5 g/kg) group, GL medium dose (7 g/kg) group, and GL high dose (14 g/kg) group. Hematoxylin-eosin staining of the prostate, and prostate blood -perfusion measured by laser speckle contrast analysis were used to evaluate the efficacy of GL. Expression of intercellular cell adhesion molecule-1 (ICAM-1) and induce nitric oxide synthase (iNOS) were determined by immunohistochemistry, and the content of interferon-γ (IFN-γ), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-10 (IL-10), chemokine ligand 1 (CXCL1) and tumor necrosis factor-α (TNF-α) were determined by electro-chemiluminescence assays. The expression of p38 mitogen-activated protein kinase (p38 MAPK), phosphatidylinositol 3-kinase (PI3K), ribosome-associated complex-alpha serine/threonine-protein kinase (Akt), nuclear factor-κ-gene binding p65 (NF-κB p65), inhibitor of NF-κB-α (IκBα), glycogen synthase kinase-3β (GSK-3β), and their phosphorylated forms were tested by Western blot.

Results: In GL, a total of 48 compounds were identified, including 14 flavonoids, 14 alkaloids, 11 carboxylic acids, 4 lactones, 2 glycosides, 2 terpenoids and 1 aldehyde. Network pharmacological analysis suggested that the mechanism of GL may be related to PI3K-Akt signaling pathway and cytokine expression. After treatment with GL, inflammatory pathological changes in the prostate of rats were significantly improved, and blood perfusion of the prostate was significantly decreased. GL reduced the expression of IFN-γ, CXCL1, TNF-α, IL-1β, iNOS, ICAM-1, p38 MAPK, p-p38 MAPK, PI3K, p-PI3K, NF-κB, p-NF-κB, IκBα, p-IκBα, GSK-3β, p-GSK-3β, p-Akt in CP rats, and increased the expression of IL-4 and IL-10 in CP rats.

Conclusion: The chemical compositions of GL were first identified. GL can improve pathological changes in the prostate and recover the prostate blood perfusion of CP rats. The possible mechanisms of GL on CP involve increasing the expression of anti-inflammatory cytokines IL-4 and IL-10, inhibiting pro-inflammatory cytokines TNF-α, IL-1β, and IFN-γ, and down regulating the expression of CXCL1, iNOS, and ICAM-1 via inhibiting PI3K-Akt and NF-κB signaling pathway.