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AIM To establish a GC method for the simultaneous determination of β-eudesmol,atractylon,atractylodin,atractylolide Ⅰ,atractylaxanthin Ⅱ and (4E,6E,12E)-tetradecene-8,10-diyne-1,3-diacetate in Atractylodes rhizome,and cluster analysis of A.rhizome according to the content level.METHODS The analysis of A.rhizome solution was performed on an HP-5 capillary column (30 m × 0.32 mm,0.25 μm) with FID as the detector,the initial temperature 100 ℃,with 10 ℃/min to 135 ℃;with 1 ℃/min to 150 ℃;with 50 ℃/min to 200 ℃ (keeping 6 minutes),and with 50 ℃/min to 250 ℃ (keeping 8 minutes).The FID detector temperature was 300 ℃ and the injector temperature was 250 ℃,with the flow rate carrier gas 1.4 L/min;The tail gas was N2 (99.999％),with the ratio of carrier gas Air ∶ H2 ∶ N2 =400 ∶ 30 ∶ 25;The sample volume was 1 μL,and the split ratio was 20 ∶ 1.The results were analyzed by cluster analysis with SPSS 21.0 statistical software.RESULTS Six constituents showed good linear relationships within their own ranges (r ＞ 0.999 6),whose average recoveries were 99.46％-100.95％ with the RSDs of 0.09-0.41.A.rhizome was divided into three categories.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of A.rhizome.
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Objective To investigate the effects of alcohol extract of bark and male flower of Eucommia ulmoides Oliv. on airway allergic inflammation induced by chicken ovalbumin (OVA) in mice; To explore its mechanism of action. Methods On day 0, day 7, mice were intraperitoneally injected OVA for sensitization, followed by nasal stimulation for 21 days to establish airway allergic inflammation mice models. The mice were divided into normal group, model group, alcohol extract of bark of Eucommia ulmoides Oliv. group, alcohol extract of male flower of Eucommia ulmoides Oliv.group,and Dexamethasone group.Each medication group was given relevant medicine for gavage. The lung tissue was embedded in HE and PAS dyeing, to observe the pathological changes of bronchus and surrounding lung. The levels of serum OVA-IgE, IL-4, IFN-γ and IL-13 were measured by ELISA. The expression of ICAM-1, VEGF, MMP9 and TIMP1 were detected by immunohistochemistry. Flow cytometry was used to detect the expression of Th17 cells in peripheral blood. The expressions of TNF-α and IL-6 mRNA in lung tissue were detected by RT-PCR. Results The model group showed changes of airway allergic inflammatory such as eosinophils and other inflammatory cell infiltration, bronchial spasm, and mucus secretion. Lung histopathology in alcohol extract of bark and male flower of Eucommia ulmoides Oliv.groups was improved significantly(P<0.05).Compared with the normal group, the levels of serum OVA-IgE, IL-4 and IL-13 increased in model group, while the level of IFN-γ decreased (P<0.05, P<0.01). The expressions of ICAM-1, VEGF and MMP9 increased, while the expression of TIMP1 decreased (P<0.01); peripheral blood IL-17+cells increased (P<0.01); the expressions of TNF-α and IL-6 mRNA increased. Compared with the model group, the levels of serum OVA-IgE, IL-4 and IL-13 decreased in alcohol extract of bark and male flower of Eucommia ulmoides Oliv. groups (P<0.05, P<0.01); the expressions of ICAM-1 and VEGF decreased (P<0.05, P<0.01); the expression of TIMP1 increased. Alcohol extract of bark and male flower of Eucommia ulmoides Oliv.could down-regulate IL-17+cells,reduce the expression of IL-6 mRNA(P<0.05,P<0.01). Alcohol extract of bark of Eucommia ulmoides Oliv. group could induce the secretion of IFN-γ (P<0.01), and down-regulate the expression of TNF-α mRNA(P<0.05).Alcohol extract of male flower of Eucommia ulmoides Oliv. group could significantly down-regulate the expression of MMP9 (P<0.05). Conclusion Alcohol extract of bark and male flower of Eucommia ulmoides Oliv.can induce the production of OVA-IgE,inhibit secretion of Th2 cytokines, inhibit the expression of adhesion molecules, depress Th17 cells, so as to inhibit the airway allergic inflammation.