| Objective:(1)the technology of ultrasonic-assisted extraction of volatile oil from aucklandiae radix was optimized.(2)to explore the differences in composition and efficacy of aucklandiae radix and its processed products,in order to expand the clinical use of aucklandiae radix.(3)to predict the analytical mechanism of aucklandiae radixin the treatment of gastrointestinal motility disorders based on network pharmacology,so as to provide a basis for further study of its molecular mechanism.Methods and Results:(1)the volatile oil of aucklandiae radix was extracted by ultrasound-assisted extraction,and the experiment was designed by single factor and Box-Behnken response surface method.taking the yield of volatile oil as the evaluation index,the best ultrasonic extraction process was determined as follows: the ratio of material to liquid was 1: 21,the temperature was 50 ℃ and the time was 36 min.Through the experimental verification of the best process,the average oil yield of volatile oil is5.60%(RSD is 1.17%),which is not significantly different from the theoretical predicted value of 5.70%,indicating that the response surface method can accurately screen the best extraction process parameters of ultrasonic-assisted extraction.(2)According to the method of Pharmacopoeia,the contents of lignin and dehydrolignolactone in different processed products of aucklandiae radix were determined.The order of the contents of hydrocarbons and dehydrolipids in aucklandiae radix and its different processed products is as follows: raw products >paper products > bran fried products > fried products > bran products.Compared with raw aucklandiae radix,the contents of aromatic hydrocarbons and dehydroamaranolide decreased after processing,but the degree of decrease was different in different processing methods.(3)the volatile components of aucklandiae radix and its different processed products were analyzed by headspace injection technique combined with GC-MS.48 kinds of volatile components were identified,accounting for 94.67% of the total,52 kinds of fried wood incense,accounting for 93.98%,41 kinds of fried wood incense of bran,accounting for 96.42%,37 kinds of stewed wood incense of bran,accounting for 96.00%,and 46 kinds of paper stewed wood incense,accounting for 94.26%.This shows that the types of volatile components of aucklandiae radix changed after processing.Among them,there are only 16 common components,including β-elemene,α-ionone,α-curcumene,β-ionone,furfural,cyclohexene,1-pentadecene,etc.,but their relative mass fractions are different.this shows that the content of these components changes or decreases or increases under the same amount of samples.The chemical constituents of volatile oil from different processed products of aucklandiae radix were analyzed by GC-MS.There were 35 kinds of wood incense,accounting for 93.58% of the total volatile oil,40 kinds of stir-fried wood incense,accounting for 94.64%,34 kinds of bran fried wood incense,accounting for 93.38%,41 kinds of bran stewed wood incense,accounting for 95.48%,and 31 kinds of paper simmering wood incense,accounting for 85.31%.There are 17 common chemical constituents in wood incense and its processed products,including β-elemene,7magenta 8-dihydroionone,trans-caryophyllene,α-ionone,geranyl acetone,α-curcumene,elemol,caryophyllin,eucalyptus-4 line 11-diene,β-eucalyptol,dehydrowood lactone and lignin lactone.The main active components of aucklandiae radix are dehydroandrographolide and hydrocarbonolactone.The content of dehydroandrographolide in the four processed products is lower than that in the raw products,and the decrease degree is different in different processed products.Compared with the raw products,the content of dehydroandrographolide in the four processed products is lower than that in the raw products.From a comprehensive point of view,different processing methods of aucklandiae radix have a certain effect on the volatile components of aucklandiae radix,which has a certain reference significance for revealing the processing law.(4)the effects of different processed products of aucklandiae radix on gastrointestinal motility in mice were investigated by using gastric emptying rate and small intestinal propulsion rate as indexes.After being treated with different processed products of wood incense,in the normal state,fried products and fried bran products have a certain inhibitory effect on gastric emptying in mice,while other processed products have no significant effect;raw products,fried products and fried bran products can promote the propulsion of small intestine in mice.Using the model of gastrointestinal motility induced by atropine in mice,except for fried products,other raw products of aucklandiae radix and their processed products could antagonize the inhibitory effect of atropine on gastric emptying.Except for the fried products of bran,other raw products of aucklandiae radix and their processed products could antagonize the inhibitory effect of atropine on the propulsion of small intestine.When neostigmine was used to induce gastrointestinal motility in mice,all the processed products of wood incense could antagonize the gastrointestinal motility excited by neostigmine,but the effects were quite different.at the same dose,the gastric residual rate of raw wood incense was the highest.the propulsion rate of small intestine was the lowest,which indicated that the antagonistic effect of raw wood incense on the exciting gastrointestinal motility of neostigmine was stronger than that of its processed products.(5)to explore the potential mechanism of aucklandiae radix in the treatment of gastrointestinal motility disorders by using the method of network pharmacology.The results showed that 97 chemical constituents,such as 2,4,Ambrosin,Methyl betulinate and so on,were active components of aucklandiae radix.One hundred and eighty-five key targets of aucklandiae radix in the treatment of gastrointestinal motility disorders were obtained,including AKT1,ALB,IL6,CASP3,SRC,TNF,MAPK1,CXCL8,HRAS,PTGS2,JUN,MMP9,IL1 B,ESR1,HSP90AA1,TLR4,KDR,MTOR,APP,MMP2 and IL2.The treatment of gastrointestinal motility disorders with aucklandiae radix involves 10 biological processes,including positive regulation of RNA polymerase II promoter transcription,regulation of cell proliferation,lipopolysaccharide-mediated signal pathway,immune response and inflammatory response,four cellular components of nucleus,extracellular space,Golgi apparatus and membrane raft,and two molecular functions of heme binding and kinase activity.There are 69 KEGG pathways involved,such as estrogen signal pathway,TNF signal pathway,cancer pathway,PI3K-Akt signal pathway,Toll-like receptor signal pathway,VEGF signal pathway,HIF-1 signal pathway and so on,of which 22 related disease pathways are involved.Conclusion:(1)Ultrasonic-assisted method is suitable for extracting volatile oil from aucklandiae radix,and it has the advantages of simple equipment and convenient operation,so ultrasonic extraction of volatile oil from aucklandiae radix has a certain value.(2)after being processed by fire,the composition and content of aucklandiae radix changed,or decreased or increased,which led to the different material basis of efficacy between raw and processed products,but in the state of inhibition and excitement,the effect of raw aucklandiae radix is better.(3)the mechanism of aucklandiae radix in the treatment of gastrointestinal motility disorders may be the joint action of the chemical components of several traditional Chinese medicine,multiple targets,multiple biological processes and multiple pathways.It reflects the characteristics of multi-components,multi-targets and multi-pathways in the treatment of diseases with traditional Chinese medicine and its compound prescription,and embodies the overall concept of traditional Chinese medicine in the treatment of diseases. |
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