Deciphering the chemical profile and pharmacological mechanism of Jinlingzi powder (金铃子散) against bile reflux gastritis using ultra-high performance liquid chromatography coupled with Q exactive focus mass spectrometry, network pharmacology, and molecular docking

doi:10.19852/j.cnki.jtcm.20230908.002

Abstract

Abstract:

OBJECTIVE: To elucidate the chemical profile and the pharmacological mechanism by which Jinlingzi powder (金铃子散, JLZP) treats bile reflux gastritis (BRG).
METHODS: A BRG model was established in rats by oral administration of the model solution. JLZP was orally administered for 35 d. Residual gastric rate and tumor necrosis factor (TNF)-α, interleukin (IL)-6, and gastrin levels in the serum were measured, and stomach tissues were collected for histopathological analysis. We used ultra-high performance liquid chromatography coupled with Q Exactive Focus mass spectrometry to identify the chemical ingredients in JLZP. Then, protein-protein interaction and herb-compound-target networks were constructed to screen potential bioactive compounds and targets. Kyoto Encyclopedia of Genes and Genomes pathway analysis was then performed to elucidate the pathway involved in the JLZP-mediated treatment of BRG. After constructing the core compound-target-pathway interaction network, molecular docking was performed to study the binding free energy of core bioactive compounds and two candidate targets [RAC-alpha serine/threonine-protein kinase (AKT1) and phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform (PIK3CA)].
RESULTS: JLZP extracts significantly promoted gastric emptying, regulating the release of cytokines (TNF-α and IL-6) and improving gastrin secretion and mucosal repair. Fifty-six compounds were tentatively characterized in JLZP. Moreover, the network pharmacology and molecular docking results showed that alkaloids and flavonoids might be the bioactive compounds in JLZP that treat BRG. JLZP might improve mucosal repair during BRG progression by modulating the phosphatidylinositol-4,5-bisphosphate 3-kinase-protein kinase B, hypoxia inducible factor-1, mitogen-activated protein kinase, forkhead box O, TNF, and IL-17 signaling pathways.
CONCLUSIONS: We elucidated the chemical constituents and the pharmacological mechanism of JLZP in treating BRG and provided a basis for clinical application.

Key words: bile reflux gastritis, liquid chromatography coupled with mass spectrometry, network pharmacology, molecular docking simulation, Jinlingzi powder

REN Hui, ZHAO Lintao, GAO Kai, YANG Yuanyuan, CUI Xiaomin, HU Jing, CHEN Zhiyong, LI Ye. Deciphering the chemical profile and pharmacological mechanism of Jinlingzi powder (金铃子散) against bile reflux gastritis using ultra-high performance liquid chromatography coupled with Q exactive focus mass spectrometry, network pharmacology, and molecular docking[J]. Journal of Traditional Chinese Medicine, 2023, 43(6): 1209-1218.

Figures/Tables 5

Figure 1 Histopathological examination of gastric antrum was observed by hematoxylin and eosin staining (×100) A: normal group; B: model group; C: domperidone group (Domperidone was administered to rats by gavage for 35 consecutive days at a dose of 3.15 mg/kg body weight); D: high dose JLZP extract group (JLZP extract was administered to rats by gavage for 35 consecutive days at a dose of 2.48 g/kg body weight); E: middle dose JLZP extract group (JLZP extract was administered to rats by gavage for 35 consecutive days at a dose of 1.24 g/kg body weight); F: low dose JLZP extract group (JLZP extract was administered to rats by gavage for 35 consecutive days at a dose of 0.62 g/kg body weight); JLZP: Jinlingzi powder.

Table 1 Comparison of residual gastric rate, and TNF-α, IL-6, and gastrin levels ($\bar{x}$ ± s)

Group	n	Residual gastric rate (%)	Serum levels (pg/mL)
Group	n	Residual gastric rate (%)	TNF-α	IL-6	Gastrin
N	10	20.64±0.14	17.25±3.80	50.71±24.99	14.79±4.96
G	10	50.53±0.24^a	32.05±15.37^a	85.19±30.87^d	8.92±1.38^a
D	10	26.14±0.15^b	19.12±7.93^b	55.52±11.10^b	13.13±4.19^b
HJ	10	26.07±0.14^b	17.63±8.94^c	46.71±25.07^c	13.07±1.91^b
MJ	10	35.50±0.20	20.12±10.10^b	63.57±23.25	12.82±2.26^b
LJ	10	40.24±0.18	22.56±7.36	42.95±28.51^c	9.74±2.85

Figure 2 Possible fragmentation pathways of alkaloids A: tetrahydroberberine; B: α-allocryptopine; RDA: Retro-Diels-Alder.

Figure 3 Core compound-target-pathway network for Jinlingzi powder in treatment of bile reflux gastritis

Figure 4 Docking simulation of key compounds to AKT1 (PDB ID: 4EKL) or PIK3CA (PDB ID: 6OAC) targets A: dihydrochelerythrine to AKT1 (Binding free energy = －8.98 kcal/mol); B: isoquercitrin to AKT1 (binding free energy = －9.76 kcal/mol); C: coptisine to AKT1 (binding free energy =－8.90 kcal/mol); D: coptisine to PIK3CA (binding free energy = －10.52 kcal/mol); E: dihydrochelerythrine to PIK3CA (binding free energy = －9.45 kcal/mol); F: jatrorrhizine to PIK3CA (binding free energy = －9.05 kcal/mol). AKT1: RAC-alpha serine/threonine-protein kinase; PIK3CA: phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform.

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