| Objective:To evaluate the in vitro anti-oxidative activity and the anti-inflammatory,antitussive and expectorant effects and its anti-inflammatory mechanism of extracts from Origanum vulgare L·Methods:(1)DPPH·free radical,hydroxyl radical(·OH)and iron ion reduction test were used to evaluate the antioxidant activity of extracts in different concentrations.UV spectrophotometry method was used to determinate the total phenols and total flavonoids content of extracts.(2)Fifty healthy mice were randomly selected by weigh,then divided in to control group(NS),Acute Bronchitis Syrup positive group(5.0mL/kg),and low?medium?high dose aqueous extract of Origanum vulgare L·treatment groups(1.35,2.70,5.40)g/kg.Then,xylenol-induced auricle swelling test,ammonia-induced cough test and bronchial phenol red excretion test were used to investigate the anti-inflammatory?antitussive and expectorant effects of Origanum vulgare L·aqueous and ethanol extrats.(3)The expression of PGE2,IL-6,TNF-?,NO,MCP-1and LDH were detected by ELISA,qPCR method was used to detect the mRNA expression levels of IL-6,TNF-?and MCP-1,in RAW264.7 macrophage inflammatory model induced by LPS to investigate the anti-inflammatory effect and mechanism of the Origanum vulgare L·aqueous extract.Results:The DPPH·Free radical scavenging rate of aqueous extract in concentration of 2mg/mL was higher than the same concentration of V_C,ethanol extract the highest concentration(2mg/mL)cleared DPPH·the ability to close to the same concentration of V_C.While Origanum vulgare L·extract reducing ability and·OH free radical scavenging ability are lower than V_C in concentration of 0.1mg/mL.The contents of total flavonoids and polyphenols in the aqueous extract of Origanum vulgare L·were 25.2?g/mg and 4.057?g/mg respectively and the content of total flavonoids and polyphenol in the ethanol extract of Origanum vulgare L·was 46.92?g/mg and 8.16?g/mg respectively.(2)Compared with the normal control group,Origanum vulgare L·aqueous and ethanol extracts were able to inhibit xylene-induced swelling of ear(P<0.05),reduce the number of cough caused by ammonia spray(P<0.05),and promote the phenol red excretion(P<0.05).anti-inflammatory,cough,expectorant effects of High and medium doses of Origanum vulgare L·aqueous extract is better than the positive control Branch syrup(P<0.05);the cough relieving,expectorant effects of high dose ethanol extract is better than the positive control medicine Branch syrup(P<0.05).(3)LDH results showed that extracts in concentration of 2.5mg/mL has toxic effect on cells,but in concentration of 0.5mg/mL,0.1mg/mL has no toxic effect to cells.Compared with LPS group,the three concentrations of aqueous extract of Origanum vulgare L·significantly inhibited the elevation of PGE2 induced by LPS.For LPS-induced TNF alpha elevation,0.5mg/mL,0.1mg/mL have inhibitory effect extracts in concentration of2.5mg/mL has significant inhibitory effect on LPS-induced IL-6 and MCP-1 and extracts in concentration of 0.5mg/ml and 0.1mg/ml has significant inhibitory effect on LPS-induced TNF alpha elevation.Conclusion:the Origanum vulgare L·has a significant antioxidant activity and anti-inflammatory,antitussive and expectorant effects,these effects may achieved by inhibit the production of PGE2,IL-6 and MCP-1. |
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