Background: Wu-Lan Thirteen-Flavour decoction (WLTd), a traditional Mongolian medicine, has been used for treating hypertension in clinical practice, but the chemical basis and underlying mechanisms remain unknown.
Methods: The main components of WLTd were identified and quantified using HPLC and UPLC-MS/MS techniques. A compound-target-disease network was constructed using network pharmacology analysis to forecast the potential anti-hypertension targets. In vivo animal and in vitro cellular experiments were performed to validate the efficacy and molecular mechanisms of renal protection of WLTd and its main active components in spontaneous hypertension.
Results: A total of 136 active compounds in WLTd were collected through relevant databases, and network pharmacology analysis identified that the aryl hydrocarbon receptor (AhR) signalling pathway may serve as a potential anti-hypertension targets. Eight of the active components, including vitexin, kaempferol, toosendanin, ursolic acid, matrine, oxymatrine, gardenoside and quercetin, were identified and quantified by HPLC and UPLC-MS/MS. WLTd effectively lowered the mean blood pressure (159.16 ± 13.91 vs 135 ± 13.37 mmHg), reduced the BUN (391.55 ± 59.96 vs 240.88 ± 51.15 mmol/L) and creatinine (1.78 ± 0.41 vs 0.67 ± 0.34 nmol/L) levels, and reduced hypertension-induced renal damage in SHR. AhR and related key gene expression changes predicted by network pharmacology analysis were validated by immunohistochemistry, RT-qPCR, and Western blot analyses. In vitro studies also showed that WLTd up-regulated AhR expression in angiotensin II-induced HEK293 cell injury.
Conclusions: Wu-Lan Thirteen-Flavour decoction effectively protects hypertension-induced renal injury by regulating Aryl Hydrocarbon Receptor signalling pathway.
Keywords: Aryl Hydrocarbon Receptor; Traditional Mongolian Medicine; Wu-Lan thirteen-flavor decoction; human embryonic kidney cell HEK293; hypertensive renal injury; molecular docking; network pharmacology.