Combining Sprague–Dawley rat uterus cell membrane chromatography with HPLC/MS to screen active components from Leonurus artemisia

Abstract

Context: Leonurus artemisia (Lour.) S.Y.Hu (Lamiaceae) (YiMuCao in Chinese) is a traditional Chinese medicine. Leonurus artemisia has been shown to have many pharmacological effects such as increasing uterine contraction amplitude, and tension, but the active components are still unknown.Objective: The objective of this study is to determine active components of L. Artemisia that are responsible for the biological activity using HPLC and cell membrane-based system.Materials and methods: The whole L. artemisia ethanol extract and its eight fractions were screened using Sprague–Dawley rat uterus cell membrane chromatography (CMC) combined with the HPLC/MS system. Oxytocin was used to investigate the activity of CMC column. The effect of active components screened from L. artemisia was studied by tension measurement of isolated rat uterine strips in vitro at a dose of 10−7–10−4 mol/L with oxytocin as a control.Results: The acetone extract showed obvious activity when compared with the eight extracts of L. artemisia. From the acetone extract, in the negative ionization mode, the active compound was identified as genkwanin, with a molecular weight of 283. In vitro pharmacological experiments proved that genkwanin promoted uterine contractions at a dose from 10−7 to 10−4 mol/L. The EC50 value was 4.86 ± 4.21 μmol/L for genkwanin and 4.30 ± 3.65 μmol/L for oxytocin on the contractile amplitude of uterine strips isolated from rats.Discussion and conclusion: Genkwanin was identified as the active compound in L. artemisia by this method. In vitro pharmacological experiments proved that genkwanin promoted uterine contractions. Genkwanin may be used to uterine inertia and may have an effect on postpartum hemorrhage.