Rapid Analysis and Guided Isolation of Astragalus Isoflavonoids by UHPLC-DAD-MSn and Their Cellular Antioxidant Defense on High-Glucose-Induced Mesangial Cell Dysfunction

Isoflavonoids, including isoflavones, isoflavans, and pterocarpans, the principal components in Astragalus membranaceus, have a great deal of versatile health-promoting benefits. In this work, as a continuation of our search for bioactive constituents from A. membranaceus, a fast high-performance liquid chromatography-diode array detection-multiple-stage mass spectrometry method was first used to analyze the isoflavonoid profile of A. membranaceus roots extract. Twelve diverse isoflavonoids in subclasses of isoflavones, isoflavans, and pterocarpans present in glycoside/aglycone pair forms were tentatively characterized; of those 12, eight major isoflavonoids were finally isolated and simultaneously quantified by the established fast UHPLC method. Furthermore, the results confirmed for the first time that Astragalus isoflavonoid aglycones could attenuate mesangial cell proliferation and extracellular matrix (ECM) accumulation triggered by high glucose levels, and the primary mechanism might be via protecting intracellular antioxidant enzymes activities and enhancing endogenous antioxidant function to lower levels of cellular oxidative damage induced by high glucose levels. Collectively, diverse Astragalus isoflavonoid antioxidants have the potential to ameliorate high-glucose-induced mesangial cell dysfunction through the regulation of cellular antioxidant defense.