ABSTRACT
INTRODUCTION: Artemisia absinthium L. is a well-known medicinal, aromatic, and edible plant with important medicinal and economic properties and a long history of use in treating liver inflammation and other diseases; however, there has been insufficient progress in quality control. OBJECTIVE: This study aimed to investigate the quality markers for the anti-inflammatory and antioxidant activities of A. absinthium based on spectrum-effect relationship analysis. MATERIALS AND METHODS: Eighteen batches of A. absinthium from different origins were used. Chemical fingerprints were obtained by ultra-performance liquid chromatography (UPLC). The chemical compositions were identified by quadrupole-Orbitrap high-resolution mass spectrometry. Anti-inflammatory activity was assessed by inhibition of cyclooxygenase-2 and 15-lipoxygenase in vitro and inhibition of nitric oxide release in lipopolysaccharide-induced BV-2 cells. Antioxidant activity was assessed by DPPH and ABTS radical scavenging assays. The relationship between bioactivity and chemical fingerprints was then analyzed using chemometrics including gray relational analysis, bivariate correlation analysis, and orthogonal partial least squares analysis. RESULTS: Different batches of A. absinthium extracts possessed significant anti-inflammatory and antioxidant activities to varying degrees. Eighty compounds were identified from A. absinthium, and 12 main common peaks were obtained from the UPLC fingerprints. P3 (chlorogenic acid), P5 (isochlorogenic acid A), and P6 (isochlorogenic acid C) were screened as the most promising active compounds by correlation analysis and further validated for their remarkable anti-inflammatory effects. CONCLUSION: This is the first study to screen the quality markers of A. absinthium by establishing the spectrum-effect relationship, which can provide a reference for the development of quality standards and further research on A. absinthium.
Subject(s)
Anti-Inflammatory Agents , Antioxidants , Artemisia absinthium , Antioxidants/pharmacology , Antioxidants/analysis , Antioxidants/chemistry , Artemisia absinthium/chemistry , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/analysis , Mice , Animals , Chromatography, High Pressure Liquid/methods , Nitric Oxide , Plant Extracts/chemistry , Plant Extracts/pharmacology , Lipopolysaccharides , Cell LineABSTRACT
A total of 29 batches of R. graveolens were used in this study, their fingerprints were obtained by ultra-performance liquid chromatography (UPLC) and their melanogenesis activities were evaluated. The common peaks were identified by quadrupole-orbitrap high-resolution mass spectrometry (Q-Orbitrap-HRMS). Eleven coumarins, six alkaloids, three flavonoids, three phenolic acids, and four other compounds were found. The spectrum-effect relationships between R. graveolens' chemical fingerprints, the melanin synthesis, and tyrosine's activation activities were established through chemometrics methods which in detail principal component analysis (PCA), gray correlation analysis (GRA), bivariate correlation analysis (BCA) and orthogonal partial least squares analysis (OPLS). The results showed that P18 (bergapten), P22 (isoimperatorin), P15 (kokusaginine), P7 (rutin), P12 (psoralen), and P13 (graveolinine) were relevant to intracellular melanin synthesis activity and tyrosinase activity. Among them, P18 (bergapten), P15 (kokusaginine), and P12 (psoralen) were validated with good melanogenesis activities. This study provides a research basis for future quality control and medicinal application of R. graveolens.
Subject(s)
Furocoumarins , Ruta , Melanins , 5-Methoxypsoralen , Furocoumarins/chemistry , Ficusin , Chromatography, Liquid , Chromatography, High Pressure Liquid/methodsABSTRACT
Daphne altaica Pall. (D. altaica; Thymelaeaceae) has long been used in traditional Kazakh medicine for the treatment of cancer and respiratory diseases. Previous studies have demonstrated the in vitro anticancer effects of D. altaica extract and its constituents in certain cancer cell lines; however, the underlying molecular mechanisms are not completely understooD. The present study aimed to investigate the molecular mechanisms underlying the activity of an ethyl acetate extract of D. altaica (DaEa) by assessing its effects on cell morphology, cell apoptosis, cell cycle progression and the expression levels of peroxisome proliferatoractivated receptor γ (PPARγ) in Eca109 cells. Cell morphology was observed under a phase contrast microscope. Cell apoptosis and cell cycle progression were assessed by flow cytometry following Annexin V/propidium iodide (PI) double staining and PI single staining, respectively. The mRNA and protein expression levels of PPARγ were determined by reverse transcriptionquantitative PCR and western blotting, respectively. Compared with the control group, the percentage of apoptotic cells, cell cycle arrest at S phase and apoptotic morphological cell characteristics were increased in DaEatreated Eca109 cells. Furthermore, DaEa treatment upregulated the mRNA and protein expression levels of PPARγ compared with the control cells. Highperformance liquid chromatography with diodearray detection indicated that daphnetin7OßDglucoside, daphnetin, demethyldaphnoretin7OßDglucopyranoside and genkwanol A were the main constituents of DaEa. Collectively, the results suggested that DaEa displayed antiproliferative activities in Eca109 cells by inducing apoptosis and S phase cell cycle arrest, as well as upregulating PPARγ expression levels.