| Objective:Atractylodes is a very important medicinal plant resource in China.Atractylodes is a dry rhizome of Atractylodeslancea(Thunb.)DC.and Atractylodes chinensis(DC.)Koidz.As a traditional Chinese medicine and ingredients for aromatized wet,it has the functions of dampness and spleen,eyesight,phlegm and cold.Clinical studies have shown that volatile oil,as the main active ingredient of Atractylodes has a variety of biological activities.Therefore,in this study,from the two aspects of extraction process and biological activity,the oil production rate,antibacterial ability,antioxidant capacity,and effect of treating spleen deficiency and dampness type irritable bowel syndrome in different areas were compared Provide theoretical basis for the development and utilization of Atractylodes resources in different producing areas and clinical rational drug useMethods:1.The volatile oil of Atractylodes was studied in four different producing areas in Jiangsu,Hubei,northeastern Chongqing and Inner Mongolia.The oil yield of volatile oil from four areas was studied by ultrasonic assisted steam distillation.The single factor(ultrasonic power,ultrasonic time,material)Based on the liquid ratio and extraction time test,the Box-Behnken response surface method was used to optimize the ultrasonic time,material-to-liquid ratio,extraction time and interaction between various factors affecting the oil yield.Process conditions.2.Escherichia coli,Bacillus subtilis,Staphylococcus aureus and Candida albicans were used as test strains.The antibacterial activity of volatile oil from 4 different areas was analyzed by filter paper method.The volatile oil of Atractylodes was determined by double dilution method.The minimum inhibitory concentration of the above tested strains was investigated to investigate the antibacterial ability of volatile oil from different habitats3.DPPH free radical scavenging method,ABTS+ˇfree radical scavenging method and hydroxyl radical scavenging method were used to compare the antioxidant activity of volatile oil from different regions of Atractylodes by using BHA standard as control.The anti oxidative activity of in vitro was studied4.The model of spleen deficiency and dampness irritable bowel syndrome was reproduced by the method of "external wetness,fatigue,diet and hunger and dysfunction".Then the rats were randomly divided into normal control group,model group,positive drug group,Jiangsu Atractylodes volatile oil group,Hubei Atractylodes volatile oil group,Northeastern Chongqing Atractylodes volatile oil group and Inner Mongolia Atractylodes volatile oil group,8 rats in each group,and treated by intragastric administration for 7 days.To explore the effects and mechanisms of volatile oil from Atractylodes on spleen deficiency and dampness type IBS in different producing areas.Determined the body weight of each group of rats,The residual rate of the stomach and the rate of small bowel advancement were measured after the last administration.The serum motilin(MTL),serotonin(5-HT)and somatostatin were detected by enzyme-linked immunosorbent assay.(SS)content changes,immunohistochemical method was used to detect the changes of IOP values of aquaporin AQP4,tight junction protein ZO-1 and Octucudin protein.Results:1.The volatile oil of Atractylodes was extracted by ultrasonic assisted steam distillation.The volatile oil of Atractylodeslancea(Thunb.)DC.was a yellowish white liquid with aromatic odor.The volatile oil of Atractylodes lancea L.was a reddish brown liquid with aromatic odor.The oil yield of Atractylodes chinensis(DC.)Koidz.was the highest in the four areas.The optimal extraction process was as follows:ultrasonic 25 min,ultrasonic power 312 W,solid-liquid ratio 1:10.5(mL/g),extraction time 2.6h.Under this condition,the measured actual extraction rate was 5.36%;the predicted extraction rate was 5.40%,and the relative error was 0.74%.2.The results of diameter of inhibition zone showed that the volatile oil of Atractylodes produced in Jiangsu,Hubei,Northeast Chongqin and Inner Mongolia had inhibitory effects on the tested strains,and the inhibitory effects of volatile oil from Atractylodes in different habitats on the same tested strains were different.The MIC range of volatile oil from Atractylodes in different producing areas was 0.79%-1.58%(?/?).Among them,the MIC value of volatile oil of Atractylodes Jiangsu against Escherichia coll,Bacillus subtilis,Staphylococcus aureus and Candida albicans was 0.79%(?/?).The MIC values of volatile oil from Atractylodes Hubei against Escherichia coll,Bacillus subtilis,Staphylococcus aureus and Candida albicans were 1.58%,0.79%,0.79%,and 1.58%(?/?),respectively.The MIC values of volatile oil from Atractylodes Chongqin against E.coll,Bacillus subtilis,Staphylococcus aureus and Candida albicans were 1.58%,0.79%,0.79,and 1.58%(?/?),respectively.The MIC values of volatile oil from Atractylodes Inner Mongolia to Escherichia coli,Bacillus subtilis,Staphylococcus aureus and Candida albicans were both 1.58%(?/?).In summary,the order of the volatile oils of Atractylodes in different areas is:Jiangsu>Hubei>?Chongqim>Inner Mongolia.3.The volatile oils of Atractylodes in four different areas all have certain antioxidant activity,and their antioxidant capacity and mass concentration have obvious dose-effect relationship.Compared with the positive control BHA standard,the ability of volatile oil from different regions to eliminate DPPH free radical scavenging ability,ABTS+ˇradicals and hydroxyl free radicals was significantly different.The IC50 values of the abalone volatile oil and the BHA control to remove DPPH radicals in the four areas were:1.992,2.408,2.392,2.856 and 1.688(mg/mL);The IC50 values of the volatiles of Atractylodes and the BHA control in the four areas were:1.178,1.712,1.816,1.987 and 0.841(mg/mL);the IC50 values of the volatile oils from the four regions of Atractylodes and the BHA control to scavenge hydroxyl radicals were:1.940,2.316,2.429,2.910 and 1.664(mg/mL).4.Compared with the normal control group,the body weight of the model group was significantly decreased(P<0.01);the residual rate in the stomach was significantly increased(P<0.01);the small intestine propulsion rate was significantly decreased(P<0.01);the serum MTL content was significantly decreased(P<0.01or P<0.05),5-HT and SS content increased significantly(P<0.05);The positive expression of AQP4 protein,ZO-1 protein and Octucudin protein in colonic mucosa was significantly decreased(P<0.01).Compared with the model group,the body weight of the rats in each administration group increased significantly(P<0.01).The gastric residual rate of the rats in the positive drug group,the volatile oil group of Jiangsu Atractylodes,the volatile oil group of Hubei Atractylodes and the volatile oil group of Northeastern Chongqing Atractylodes was significantly increased(P<0.05orP<0.01);The positive intestine rate of the positive drug group,Jiangsu Atractylodes volatile oil group,Hubei Atractyl-odes volatile oil group and Chongqing Atractylodes volatile oil group was signi-ficantly decreased(P<0.01).The serum MTL content of rats in each administr-ation group increased significantly(P<0.01orP<0.05).The positive drug group,Jiangsu Atractylodes volatile oil group,Hubei Atractylodes volatile oil group,and the Northeastern Chongqing volatile oil group rats showed a significant de-crease in serum 5-HT content and SS content(P<0.01);ZO in colonic mucosa of rats in each administration group The-1 protein IOD value was significantl-y increased(P<0.01).The AQP4 protein and Occuludin protein IOD values 1-n the colonic mucosa of the positive drug group,the Jiangsu Atractylodes vol-atile oil group,the Hubei Atractylodes volatile oil group,and the Northeastern Ch-ongqing Atractylodes volatile oil group were significantly increased(P<0.01).Conclusions:1.The volatile oil extraction process model of Atractylodes established by r-esponse surface method has high oil yield,short time-consuming,simple opera-tion and good prediction of test results.2.The volatile oils of Atractylodes in Jiangsu,Hubei,Northeastern Chongqing and Inner Mongolia have inhibitory effects on Escherichia coli,Bacillus subtilis,Staphylococcus aureus and Candida albicans,and the inhibitory effects o-f volatile oil of Atractylodes from different habitats on the same tested bacteria are different.The order of total bacteriostatic activity was:Jiangsu>Hubei?Northeastern Chongqing>Inner Mongolia.It can be used for the evaluation of antibacterial effect of Atractylodes medicinal herbs and the screening of natural bacteriostatic agents.3.The volatile oils of Atractylodes in four different areas have the ability to scavenge DPPH free radicals,ABTS+ˇfree radicals and hydroxyl radicals,and increase with the increase of mass concentration.The antioxidant activities of volatile oils from different regions have different.The order of total antioxidant activity is:BHA>Jiangsu>Hubei>?Northeastern Chongqing>Inner Mongolia.It can be used for the evaluation of antioxidant activity of Atractylodes medicinal materials and the screening of natural antioxidants.4.The volatile oil of Atractylodes in four different producing areas may be up-regulated by MTL levels in the serum of spleen-deficiency type IBS rats,down-regulating 5-HT,SS levels and up-regulating AQP4 protein,Occludin protein and ZO-1 protein in rat colon tissue.Expression and treatment of spleen deficiency and dampness-type irritable bowel syndrome. |
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