RESUMEN

This study aimed to evaluate the anti-inflammatory mechanism of neferine, a bisbenzylisoquinoline alkaloid, on the lipopolysaccharide (LPS)-induced inflammation in Human Umbilical Endothelial Cells (HUVECs) and pulmonary aorta cells. Production of pro-inflammatory cytokines and nitric oxide (NO) was determined using Griess reaction in human endothelial cells. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and mitogen-activated protein kinases (MAPKs) were analyzed using real time PCR and western blotting. Neferine significantly prevented the NO, TNF-α, COX-2, iNOS, IL-1B, and other inflammatory mediators formation in increasing dose as compared to LPS-induced human endothelial cells. The expressions of NADPH oxidase subunits p22phox, p47phox, and gp91phox were increased in LPS-induced HUVECs but neferein was able to reverse the effect in a dose-dependent manner. The anti-inflammatory effects of neferine in LPS-induced endothelial cells are attributed through the modulation of MAPK and NF-κß pathways. Collectively, these results suggest that neferine could be beneficial in the early treatment of atherosclerosis to prevent stroke and heart disease.

Asunto(s)

Antiinflamatorios/farmacología , Bencilisoquinolinas/farmacología , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Inflamación/tratamiento farmacológico , Animales , Antiinflamatorios/administración & dosificación , Aorta/citología , Aorta/patología , Aterosclerosis/tratamiento farmacológico , Aterosclerosis/patología , Bencilisoquinolinas/administración & dosificación , Células Cultivadas , Ciclooxigenasa 2/metabolismo , Citocinas/metabolismo , Relación Dosis-Respuesta a Droga , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Inflamación/patología , Mediadores de Inflamación/metabolismo , Lipopolisacáridos/toxicidad , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Ratas , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Our work focuses on the quality control and structural identification of Photocyanine as a cancer therapeutic photosensitizer. Photocyanine is a mixture which contains four ZnPcS2P2 type substituted Phthalocyanine isomers. In order to obtain the single component from Photocyanine, the mixture of four isomers possessing the similar structures and chemical property had been isolated and purified. An HPLC method with a mixture of methanol-acetonitrile-ion-pair buffer as the mobile phase was applied to isolate the four isomers by means of a semi-preparative C18 column. To remove the salts which were mixed in the preparative product, a SPE C18 column was used to separate the salts by elution with water and then the marker component was eluted by methanol. Subsequently, a column of Sephadex LH-20 gel was applied to elute the crudes with methanol to desalination. The purity of the isolated compound was measured by TLC and four different isomers of phthalocyanine were obtained. The chemical structures of them were elucidated by 1H NMR spectra, gCOSY and NOE1D. An HPLC-DAD method was developed for simultaneously determination of four major isomers in Photocyanine with a C18 column (Grace Smart, 150 mm x 4.6 mm ID, 5 microm). The separation was carried out with a gradient program at a flow rate of 1.0 mL x min(-1). The mobile phase was a mixture of acetonitrile and ion-pair buffer (0.01 mol x L(-1) hexadecyl trimethyl ammonium bromide and 0.01 mol x L(-1) potassium dihydrogen phosphate, adjusted the pH value to 6.8 with potassium hydroxide solution). The resolution values of four isomers were 2.5, 1.20, 1.33, and 1.8. Linear regression analysis for four compounds was performed by the external standard method. Four constituents were linear in the concentration range of 0.005 to 10 microg. The values of relative standard deviation (RSD) of intra-day were 0.12%, 0.66%, 0.99%, and 1.21%, respectively. The limits of detection for four compounds were 15 ng, 20 ng, 12 ng, and 25 ng, respectively. This method was simple, accurate and reproducible. The developed method can be successfully applied to analyze isomers in Photocyanine.

RESUMEN

The methods of 1H Nuclear Magnetic Resonance (NMR) and mass spectroscopy were used in detecting the metabolites of brodimoprim (BDP) in rat plasma. The endogenic compounds in the plasma were removed with solid phase extraction (SPE) column firstly, then the mixture of metabolites was identified with 1H NMR and mass spectroscopy (MS). Two metabolites of BDP in the plasma at 20h were detected, they were demethyl-brodimorpim glucuronide and brodimoprim sulfurate. The study proved that the method of SPE coupled with NMR and MS can be applied to the analysis of metbolites in plasma quickly and conveniently.