RESUMEN
The present work is to establish an HPLC characteristic chromatograms of Asarum heterotropoides var. mandshuricum(AH) and A. sieboldii(AS), combined with cluster analysis for the identification of the two species, and predict their potential anti-inflammatory related targets by network pharmacological method. Eighty-nine samples(12 batches of AS and 77 batches of AH) were analyzed, and 11 characteristic peaks were identified by reference substances, UV spectrum and LC-MS. Cluster analysis showed that AS and AH were divided into two groups, and the ratio of characteristic peak areas can be used to distinguish them. When the ratio of characteristic peak sarisan to kakuol was greater than 5, it was AS, and when the ratio was less than 2, it was AH. The network pharmacological analysis of 119 constituents of Asari Radix et Rhizoma suggested that the anti-inflammatory effect of Asari Radix et Rhizoma might be related to COX-2, COX-1, iNOS, MAPK14, NR3 C1, PPARG and TNF. Among them, COX-2 is a relatively key target, which interacted with the characteristic constituents, asarinin, sesamin, safrole, methyleugenol and sarisan. The characteristic constituents asarinin and sesamin also interacted with the iNOS and MAPK14. Safrole and sarisan can also interact with iNOS, COX-1 and LAT4 H. Methyleugenol also showed interaction with COX-1 and LAT4 H. Since asarinin and sesamin interacted with three targets, COX-2, iNOS and MAPK14, it implied that they were the main active constituents for the anti-inflammatory activity of Asari Radix et Rhizoma. The COX-2 inhibitory activities of asarinin and sesamin were further studied by molecular docking and bioassay. The HPLC method established was simple, feasible and reliable, with predicted anti-inflammatory targets and anti-inflammatory constituents, which could provide a reference for improving the quality evaluation system of Asari Radix et Rhizoma.
Asunto(s)
Antiinflamatorios/aislamiento & purificación , Asarum/química , Cromatografía Líquida de Alta Presión , Simulación del Acoplamiento Molecular , Fitoquímicos/aislamiento & purificación , Rizoma/químicaRESUMEN
UbcH5c belongs to the ubiquitin-conjugating enzyme family and plays an important role in catalyzing ubiquitination during TNF---triggered NF-B activation. Therefore, UbcH5c is a potent therapeutic target for the treatment of inflammatory and autoimmune diseases induced by aberrant activation of NF-B. In this study, we established a stable expression system for recombinant UbcH5c and solved the crystal structure of UbcH5c belonging to space group P222with one molecule in the asymmetric unit. This study provides the basis for further study of UbcH5c including the design of UbcH5c inhibitors.
RESUMEN
The aim of the study was to prepare and evaluate the quality of negatively charged selfmicroemulsifying drug delivery system of curcumin (NC-CUR-SMEDDS).Based on the CUR-SMEDDS,the optimum amount of excipients was confirmed by the single-factor design experiment taking mean particle size,Zeta potential,drug entrapment efficiency and the drug loadings of curcumin as the evaluation indices for the preparation of NC-CUR-SMEDDS.The quality of NC-CUR-SMEDDS was evaluated by observing its appearance status,transmission electron microscope micrographs and determining particle diameter,Zeta electric potential,drug entrapment efficiency and drug loading.As a result,it was found that the Zeta potential reached -43.43 ± 0.29 mV when 4％ docusate sodium was added.The appearance of NC-CUR-SMEDDS remained clarified and transparent,and the microemulsion droplets appeared spherical without aggregation with uniform particle size distribution.The mean particle size was 14.08 ± 0.082 nm,the drug loading of curcumin was 26.48 mg·g-t and the drug entrapment efficiency was 94.12％.It was concluded that NC-CUR-SMEDDS with high entrapment efficiency and uniform particle size distribution met the requirement of inflammatory target binding in the colon.