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Aim To investigate the mechanism and search for potential biomarkers of ovalbumin ( OVA ) -induced asthma in mice base on lipidomics. Methods A BALB/c mouse model of asthma was prepared by OVA. TNF-α, IL-4, IL-10, IFN-γ levels in BALF and IgE level in serum were measured by ELISA. The inflammatory changes in mouse lung tissue were observed using HE staining. Lipid mediators ( LMs) in lung tissue and serum were quantified with UPLC-MS/ MS strategy. Results IgE level in serum and TNF-α, IFN-γ levels in BALF were higher (P <0.05) of asthmatic mice.Typical inflammatory manifestations were seen in lung tissue of asthmatic mice. A total of 57 lipid mediators were quantified with UPLC-MRM. LMs metabolic profiles differed significantly in serum and lung tissue between asthmatic and normal mice, 17 significantly different LMs were found in lung tissue and 6 LMs were found in serum, and the differential metabolites were produced through the cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 oxidase (P450) metabolic pathways. Conclusions OVA-induced allergic asthma can cause disorder of lip-id mediators, LMs and cytokines are involved in the occurrence and development of asthma. The differential LMs have potential research value as biomarkers for the development of allergic asthma.
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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.
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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.
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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.