ABSTRACT
OBJECTIVE:To conduct component analysis for the 40％ ethanol eluate of Hyssopus officinalis,and investigate its improvement effect on inflammation in asthmatic mice. METHODS:The 40％ ethanol eluate of polyamide resin column was col-lected,and HPLC-high resolution mass spectrometry was used for the component analysis of 40％ ethanol eluate of H. officinalis. Totally 72 mice were randomly divided into blank group (normal saline),model group (normal saline),dexamethasone group (positive control,1.6 mg/kg) and 40％ ethanol eluate of H. officinalis high-dose,medium-dose,low-dose groups (200,100,50 mg/kg),12 in each group. Except for normal group,mice in other groups were intraperitoneally injected 0.2 mL ovalbumin (OVA)for sensitization in 0,14 d and intragastrically administrated in 25-31 d,once a day. After administration,2 mg/mL OVA was dropped in nose for 7 d. After 24 h of last dropping in nose,tumor necrosis factor α(TNF-α),interleukin-4(IL-4),interfer-on-γ(IFN-γ)levels in bronchoalveolar lavage fluid(BALF)were detected;pathological changes in lung tissue were observed. RE-SULTS:Totally 11 compounds were identified,the relative percentage content of 40.89％. The main components were rosmarinic acid,luteolin 7-O-β-D-rhamnoserhamnose (1→6)-α-D-pyran glucoside,hyperoside,etc. Compared with blank group,TNF-α, IL-4 levels in BALF in model group were increased,IFN-γ level was declined,and IL-4/IFN-γ ratio was enlarged(P<0.01);lung tissue was seriously damaged,there was infiltration of inflammatory cells around the blood vessels. Compared with model group, TNF-α,IL-4 levels in BALF in dexamethasone group,40％ ethanol eluate of H. officinalis high-dose,medium-dose groups were declined,IFN-γ level was increased,and IL-4/IFN-γ ratio was reduced (P<0.01);pathological changes in lung tissue were improved. CONCLUSIONS:The established analysis method can effectively analysis the chemical components of 40％ etha-nol eluate of H. officinalis,which has certain regulatory effect on releasing inflammatory factors and reducing inflammatory lesions in lung tissue of mice with bronchial asthma.
ABSTRACT
OBJECTIVE:To provide reference for clarifying the pharmacodynamic material basis of anti-inflammation effect of Uygur medicine Hyssopus cuspidatus. METHODS:HPLC was conducted to establish the fingerprint of 8 extracts with different po-larities. Using macrophage RAW264.7 as object,the inhibitory effect of extracts with different polarities on inflammatory factor ni-tric oxide(NO),TNF-α and IL-6 in supernatant of cell culture medium was detected,the differences of anti-inflammatory activity in vitro were compared. Grey correlation analysis method was used to analyze the spectrum-effect relationship of peak area of common peaks and anti-inflammation activity. RESULTS:The fingerprint of 8 extracts with different polarities showed obvious differences. Anti-inflammation in vitro results suggested that 30% ethanol extract had the strongest inhibitory effect on inflammatory cytokines in vitro. There were 14 common peaks in the established HPLC fingerprints,the 5 common peaks that was closely related to the in-hibitory effect of inflammatory factors in vitro were peak 8,6,5,10,13,respectively;peak 8 was rosmarinic acid. CONCLUSIONS:The established fingerprint and its spectrum-effect relationship with anti-inflammation activity in vitro can provide certain reference for its pharmacodynanic material basis study.
ABSTRACT
With development of bio-technique, more and more proteins were applied as clinical approaches. However, the protein homogeneity, especially the N-glycosylation limited the further research and application of these protein drugs. The analysis method for N-glycans is believed to be critical in protein drugs development. To enhance the N-glycans isolation efficiency and accelerate the pretreatment, a new strategy was built on ultrafiltration-devices. New methods increased the isolation efficiency of N-glycans containing N-acetylglucosa mine with 10%-20%. The degrading of N-glycans containing sialic acids was also minimized with this method. 20%-100% more N-glycans with sialic acids were isolated. The pretreatment was finished within 30 min. Coupled with HPLC-HRMS, an effective and reliable strategy designed for protein drugs N-glycans analysis were developed.
ABSTRACT
With development of bio-technique, more and more proteins were applied as clinical approaches. However, the protein homogeneity, especially the N-glycosylation limited the further research and application of these protein drugs. The analysis method for N-glycans is believed to be critical in protein drugs development. To enhance the N-glycans isolation efficiency and accelerate the pretreatment, a new strategy was built on ultrafiltration-devices. New methods increased the isolation efficiency of N-glycans containing N-acetylglucosa mine with 10%-20%. The degrading of N-glycans containing sialic acids was also minimized with this method. 20%-100% more N-glycans with sialic acids were isolated. The pretreatment was finished within 30 min. Coupled with HPLC-HRMS, an effective and reliable strategy designed for protein drugs N-glycans analysis were developed.