A solid-phase microextraction method is introduced to overcome limitations of classical phytochemical pattern of identifying bioactive compounds, including tedious and time-consuming separation and purification step and consumption of large amounts of organic solvents , which was non-environmentally- friendly. In this proposed method for solid-phase microextraction, polyvinylidene fluoride [email protected] @ [email protected] as a solid part of the extractors were pushed into sample solution of medicinal plants , and the procedure was followed by stirring and easily dissociation of receptor binding ligands in organic solvent through pulling out of the functionalized fibers. Xanthine oxidase was chosen as the model receptor, while isoacteoside was selected as the model inhibitor. Several effecting parameters were optimized by experimental design, including temperature, ion strength and pH. Nine bioactive components were obtained from extract of Plantago depressa by using the established solid-phase micro-extraction method. The limit of detection and limit of quantification of the nine components ranged from 0.0008 to 0.03 mg mL −1 and from 0.001 to 0.016 mg mL −1 , respectively. The RSD values of intra-day and inter-day precisions ranged from 0.24% to 2.19% and 0.62%–2.84%, respectively. The average recoveries of the nine components were from 95.06 to 104.03% with relative standard deviation (RSD) values from 1.02 to 2.90% for Plantago depressa . The RSD values of stability of the nine components ranged from 1.36% to 2.74%, which satisfied the requirements of an analytical method. In addition, surface plasmon resonance biosensor was utilized to corroborate the binding affinity between these compounds and receptor. The avidity values of these ligands corresponded well with their IC 50 values. The results confirmed that polydopamine and polyethyleneimine hybrid modified polyvinylidene fluoride fibers based solid-phase microextraction method was successfully utilized for locating bioactive compounds of medicinal plants.
Yanan Wu,Yihan Yang,Liping Du,Yibin Zhuang,Tao Liu
Glycosylation reactions mediated by UDP-glycosyltransferases (UGTs) are common post-modifications involved in plant secondary metabolism and significantly improve the solubility and bioactivity of aglycones. Penstemon barbatus is rich in phenylethanoid glycosides (PhGs), such as echinacoside and verbascoside. In this study, a promiscuous glycosyltransferase UGT84A95 was identified from P. barbatus. In vitro enzyme assays showed that UGT84A95 catalyzed the glucosylation of the phenol hydroxyl group of PhGs efficiently as well as other structurally diverse phenolic glycosides, including flavonoids, terpenoids, stilbene glycosides, coumarins, and simple polyphenols. By using UGT84A95, 12 glycosylated products were prepared and structurally identified by NMR spectroscopy, among which 7 are new compounds. These findings suggest that UGT84A95 could be a potential biocatalyst to synthesize multi-glycosylated glycosides.
Ethnopharmacological relevance The Chinese herbal Plantaginis Herba (PL) is one of the most widely used plants for both medicinal and dietary purposes. Plantaginis Herba is the main medicine used in a traditional Chinese prescription called Cheqiancao decoction, and it is known for its liver and kidney protective properties. Aim of the study : The aim of the present study was to explore the interventions and mechanisms of PL in ADR nephropathy by performing an integrated analysis of in vitro and in vivo experiments. Materials and methods The ingredients of PL were analysed by Ultra High Performance Liquid Chromatography (UPLC). The biochemical indicators of renal injury in the serum and urine were detected by a Micronumerase assay and ELISA. The renal histopathology and ultrastructure were analysed by H&E staining, Masson’s trichrome staining and transmission electron microscopy (TEM), respectively. By identifying the targets of PL and ADR nephropathy, a network of PL–constituents–targets–ADR nephropathy was constructed, and a KEGG signaling pathway enrichment analysis was performed to complete the network pharmacology. A transcriptomic analysis was performed on a sequencing platform (Illumina). Results Plantaginis Herba significantly decreased the levels of BUN, Scr, ALB, MAU and KIM-1. Plantaginis Herba inhibited renal histopathological injury and alleviate foot process fusion and podocyte basement membrane thickening. In addition, the results of the transcriptomic analysis and network pharmacology analysis indicated that the HIF-1, TGF-β, and PI3K/AKT signaling pathways; apoptosis; and ECM–receptor interactions might be pivotal pathways for the effect of the PL intervention on ADR nephropathy. Moreover, the validation results revealed that PL could effectively attenuate collagen fibre deposition and inhibit oxidative stress. Plantaginis Herba could regulate the expression levels of pivotal proteins in the β-catenin/TGF-β1, HIF-1, and PI3K signaling pathways in renal tissues. Plantaginis Herba could reduce the level of apoptosis and the percentage of decrease in the mitochondrial membrane potential (MMP) in primary renal cells from rats with ADR nephropathy, and regulate key proteins involved in mitochondrial apoptosis. Furthermore, Luteolin from PL had good affinity for HIF-1α, and the ability of Luteolin to ameliorate in ameliorating ADR-induced MPC-5 cell injury was attenuated by overexpressing HIF-1α. Conclusions Plantaginis Herba alleviates ADR-induced nephropathy by regulating mitochondrial apoptosis via the HIF-1α signaling pathway. Luteolin may be one of the active ingredients responsible for these effects, and these findings provide an innovative strategy for the intervention and treatment of ADR nephropathy.
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