| Background:Chimonanthus is a genus of the genus Chimonanthus,which is widely distributed in China.The resources of the genus Chimonanthus in Shiyan are very rich.Previous studies have found that the extracts of various species of Chimonanthus have anti-inflammatory effects.Objective:To study the anti-inflammatory effect and mechanism of Chimonanthus flowers,and to provide scientific basis for the development and utilization of Chimonanthus praecox flowers resources in Shiyan region,this study uses the essential oil extracted from Chimonanthus praecox flowers in Shiyan region for anti-inflammatory experiments,to study the protective effects and the mechanism of essential oil extracted from Chimonanthus praecox flowers on inflammatory rats by detecting relevant indicators.Methods:(1)Seventy-two SPF SD rats were randomly divided into blank control group,model group,positive control group(aspirin 200 mg/kg),essential oil low,medium and high concentration groups.The blank control group and the model group were treated with distilled water by gavage.The positive control group was treated with aspirin by gavage.The low,medium and high concentration of the essential oil groups were given 1%、3%、8% of essential oil extracted from Chimonanthus praecox flowers at 10 m L/kg.All groups were gavaged once a day.On the 5th day of the experiment,30 minutes after gavaged,0.1 m L Freund’s complete adjuvant was injected subcutaneously into the right foot plantar of each rats of groups,and the blank control group was subcutaneously injected with 0.1 m L normal saline.Observe and measure the toe volume of rats before and 1 hour,2 hours,3 hours,5 hours,24 hours,2days,3 days after inflammation by drainage method,and calculate the toe’s swelling rate of each group of rats at each time point.After each group of rats was anesthetized,blood was taken from the heart,and lumbar of spinal tissue and swelling tissues of foot were taken to detect the levels of TNF-α,IL-1βand other related inflammatory factors,and Western Blot method was taken to detect the key proteins of nuclear factor kappa-B passway.Each group of rats was perfused after anesthetized,and the local tissues of the spinal cord lumbar and foot swelling were fixed and paraffin-embedded,and pathological sections were made.The microglial cells in spinal cord tissue were labeled with anti Iba1 and observed with a confocal microscope to calculate the activation of microglia in lumbar segment of the spinal cord tissue.(2)Fifty SPF-grade KM mice were randomly divided into a model group,a positive control group(aspirin 200 mg/kg),and essential oil low,medium,and high concentration groups.The model group was gavaged with distilled water,and the positive control group was gavaged with aspirin.The low,medium,and high concentration groups of essential oil were gavaged with 1%、3% and 8% of essential oil extracted from Chimonanthus praecox flowers at 10 m L/kg.All groups were gavaged once a day.On the seventh day of the experiment,30 minutes after gavaged,20 μL of xylene was smeared to the right ear of each group’s mice.After half an hour,the mice were sacrificed by cervical dislocation,and both sides of the ear of all the mice of each group were punched with a 6mm diameter punch.To weigh the round ears pieces of each side of every mouse with a precision balance,and calculate the degree of the ear’ swelling and the swelling suppression rate of the mice in each group.Results:The medium and high concentration groups of essential oil had a significant inhibitory effect on toe’ swelling in rats at 1 hour,2 hours,3 hours,5hours,24 hours,and 2 days after inflammation with Freund’s complete adjuvant(P<0.05).The medium and high concentration groups of essential oil can significantly reduce the content of GM-CSF in serum(P<0.01),while the effect of low concentration group is not obvious;however,the impact of each groups of essential oil on IL-1 β,TNF-α in serum is not obvious(P>0.05).Compared with the model group,p-NF-κB p65 in the spinal cord tissue of each concentration group of essential oil was significantly reduced,and the difference was statistically significant(P<0.01).The content of TNF-α in the swelling tissue of the foot of each concentration group of essential oil was significantly reduced,and the difference was statistically significant compared with the model group(P<0.01);The content of IL-1β in the swelling tissue of the foot of mid and high concentration groups were significantly reduced,and the difference was statistically significant compared with the model group(P<0.01 or P<0.05).The activation rate of microglia in the spinal cord tissue of the high concentration group of essential oil was significantly reduced,and the difference was statistically significant compared with the model group(P<0.01).Each concentration group of essential oil can alleviate the infiltration of inflammatory cells in the swelling tissue of the foot in different degrees.High concentration of essential oil group has a significant inhibitory effect on ear’swelling induced by xylene in mice(P<0.01),but low and medium concentration groups’ effects were not significant.Conclusion:Essential oil extracted from Chimonanthus praecox flowers has a significant protective effect on mice inflammed by Freund’s complete adjuvant.Its mechanism may be related to the down-regulation of the expression of TNF-α,IL-1β and other proteins through NF-κB pathway,but the effects on TNF-α and IL-1βin serum is not obvious.The high concentration group can significantly reduces mice’ ear’ swelling caused by xylene,but its anti-inflammatory effect is weaker than aspirin. |
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