Exploration of the chemical constituents and mechanisms of Runzaoling prescription in treatment of Sjögren's syndrome using UHPLC-Q-Exactive Orbitrap MS/MS combined with experimental validation

Abstract

Background

Runzaoling prescription (RZL) has been used clinically for many years, and has shown potential therapeutic effects in previous studies for the treatment of Sjögren's Syndrome (SS). Even so, the specific active components and underlying mechanisms by which RZL exerts its anti-SS effects remain unclear.

Materials and methods

Ultra-high performance liquid chromatography coupled with Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-Exactive Orbitrap MS/MS) was employed to systematically identify the chemical constituents of RZL in vitro, as well as those components that enter the bloodstream in mice. The main active components, core genes and related signaling pathways of RZL for SS were screened by network pharmacology. Furthermore, the affinity between important components and their primary targets was predicted using molecular docking technology. Finally, the animal model of spontaneous SS (NOD mice) was established to evaluate the efficacy of RZL.

Results and conclusions

728 components were identified in RZL, and 49 prototype components as well as 7 metabolites were identified from the blood of mice. Network pharmacological analysis showed that RZL targeted key proteins like ICAM1, TNF, TLR4, and EGFR through active components such as isorhapontigenin, hypaphorine and cyanidin, modulating pathways like NF-κB, PI3K/AkT, MAPK, HIF-1 and TNF to exhibit the anti-inflammatory effects for treating SS. Molecular docking revealed that key active components of RZL effectively interact with proteins in the NF-κB pathway. Animal studies indicated that RZL alleviated pathological damage, enhanced saliva production and submandibular gland index in SS model mice. Additionally, it may regulate the TLR4/MyD88/NF-κB signaling pathway and effectively inhibit the inflammatory response.