Study On The Effect Of Fried Rice On The Chemical Composition And Efficacy Of Codonopsis Pilosula

Objective:Codonopsis pilosula is a traditional tonic Chinese medicine.Fried Codonopsis pilosula with rice is a commonly used processed product.It has a significant effect on the treatment of spleen deficiency and is included in Chinese Pharmacopoeia.In order to clarify the types of rice used in the preparation of rice-fried Codonopsis pilosula and clarify their characteristics,this study compared and analyzed the main chemical composition content of Codonopsis pilosula and different types of rice-fried Codonopsis pilosula,fingerprint fingerprints,and the effect on spleen-deficiency rats and intestinal flora.The similarities and differences in the ingredients and efficacy of different rice-fried Codonopsis pilosula lay the foundation for enriching the connotation of rice-fried Codonopsis pilosula and perfecting the processing standards.Methods:1.Comparison of the main chemical components of Codonopsis pilosula and different rice-fried Codonopsis pilosulaThe content of alcohol-soluble extracts was determined by hot leaching method,5-hydroxymethylfurfural(5-HMF)and Lobetyolin content were determined by HPLC method,and Codonopsis pilosula polysaccharide content was determined by phenol-concentrated sulfuric acid method.Compare the chemical composition.2.Research and establishment of fingerprints of Codonopsis pilosula and different rice stir-fried Codonopsis pilosulaUltra-high performance liquid chromatography(UPLC)method was adopted,the chromatographic column was Agilent Poroshell SB-C18,the mobile phase was acetonitrile-0.15%glacial acetic acid water(gradient elution),the flow rate was0.3m L/min,the detection wavelength was 267nm,the column temperature It is 30°C and the injection volume is 1μL.Using Lobetyolin as a reference,UPLC fingerprints of Codonopsis pilosula,Millet Fried Codonopsis pilosula,and Rice Fried Codonopsis pilosula(9 batches each)were established,and the similarity evaluation was carried out using the"Chinese Medicine Fingerprint Similarity Evaluation System(2012Edition)",using SPSS 20.0 The software performs cluster analysis and principal component analysis.Based on the above chromatographic conditions,using UPLC-Q-Exactive-MS/MS liquid mass spectrometry technology,The constituents of Codonopsis pilosula were rapidly isolated and identified by HRMS in the positive and negative ion mode using both full scan and two stage threshold-triggered mass modes.3.Pharmacodynamic research of Codonopsis pilosula participating in different rice frying in rats with spleen deficiencyThree factors of rhubarb suffering from bitter cold,exhaustion of swimming,and eating disorder were used to induce SD rats to cause spleen deficiency model.Gavage Codonopsis pilosula,millet-fried Codonopsis pilosula and rice-fried Codonopsis pilosula medicinal liquid,respectively,to increase the weight of rats with spleen deficiency,the length of the last swim,the content of CK,the excretion rate of D-xylose,the intestinal propulsion rate,GAS,MTL,VIP,SS and other gastrointestinal function related indicators,spleen index,thymus index,total number of white blood cells,CD4~+/CD8~+ratio,Ig G,TNF-αand IL-6 content and other immune function related indicators,AQP 3 and Na~+-K~+-ATPase content Test the related indicators of water and liquid metabolism to compare the similarities and differences of the efficacy of different rice-fried Codonopsis pilosula.4.Analysis of the difference of intestinal flora between Codonopsis pilosula and different rice-fried Codonopsis pilosulaThe experimental animal model and the method of administration are the same as in"3.Efficacy test".On the last day of administration,feces samples of each group of rats are collected,and total DNA of feces samples was extracted,and primers were designed for amplification according to the variable region of bacterial 16S r RNA V3～V4.The Illumina Miseq platform was used for high-throughput sequencing to compare and analyze the abundance and diversity of bacterial community composition.Results:1.Comparison of the main chemical components of Codonopsis pilosula and different rice-fried Codonopsis pilosula(1)Alcohol-soluble extractThe content of the alcohol-soluble extracts of Lu Codonopsis pilosula is in the order of stir-fried Codonopsis pilosula with rice>Fried Codonopsis pilosula with millet>Codonopsis pilosula;verified by different sources,the content of the alcohol-soluble extracts of Codonopsis pilosula and Codonopsis tangshen Oliv.are in the order of stir-fried Codonopsis pilosula with millet>Fried Codonopsis pilosula with rice>Codonopsis pilosula.,The trend of change is inconsistent.(2)5-HMFThe 5-HMF content of Codonopsis pilosula is in the order of rice-fried Codonopsis pilosula>Millet-fried Codonopsis pilosula>Codonopsis pilosula;after verification of different sources,the 5-HMF content of plain flower Codonopsis pilosula is consistent with that of Codonopsis pilosula,and the 5-HMF content of Codonopsis tangshen Oliv.is millet-fried Codonopsis pilosula≈fried Codonopsis pilosula with rice>Codonopsis pilosula,the change trend is not consistent.(3)Codonopsis pilosula polysaccharidesThe polysaccharide content of Lu Codonopsis pilosula is in the order of rice-fried Codonopsis pilosula>Codonopsis pilosula>millet-fried Codonopsis pilosula;the change trend is the same after verification by different sources.(4)LobetyolinThe content of Lobetyolin in the order of Codonopsis pilosula>Fried Codonopsis pilosula with millet>Fried Codonopsis pilosula with rice;After verification from different sources,the content of Codonopsis pilosula and Codonopsis tangshen Oliv.are in the order of millet-fried Codonopsis pilosula>Fried Codonopsis pilosula with rice>Codonopsis pilosula has inconsistent trends.2.Research and establishment of fingerprints of Codonopsis pilosula and different rice stir-fried Codonopsis pilosulaBy optimizing the preparation conditions and chromatographic conditions of the test solution,the UPLC characteristic maps of Codonopsis pilosula,millet-fried Codonopsis pilosula,and rice-fried Codonopsis pilosula were established,and methodological investigations were carried out to prove that the method is stable and feasible.The UPLC fingerprints of 9 batches of Codonopsis pilosula products have 6common peaks.Millet fried Codonopsis pilosula and rice fried Codonopsis pilosula each have 10 common peaks,and the similarities are both greater than 0.90.Two common peaks of 5-HMF and Lobetyolin are identified.The UPLC fingerprint of Codonopsis pilosula"rice"has obvious common peak characteristics,and the peak areas of chromatographic peaks 3,5,6,and 7 are significantly increased.Compared with the fingerprints of millet fried Codonopsis pilosula,the fingerprints of rice fried Codonopsis pilosula are basically the same except for peaks 6,7 Except for the same,the peak areas of other chromatographic peaks are relatively large.The results of cluster analysis and principal component analysis showed that 9 batches of Codonopsis pilosula were clustered into one category,while millet-fried Codonopsis pilosula and rice-fried Codonopsis pilosula clustered into another category.Using UPLC-Q-Exactive-MS/MS technology,combined with database and literature,a total of 41 chemical components were identified,mainly lactones,organic acids and phenylpropanoids,including abscisic acid,syringaldehyde,and osmanthus Oxalin and caffeic acid are unique components in Codonopsis pilosula;lactose and 6-gingerol are uniquecomponentsinmilletfriedCodonopsispilosula;(+)-pinoresin-β-D-glucopyranoside,quercetin,adenine,Luteolin only exists in Codonopsis pilosula and rice-fried Codonopsis pilosula;5-hydroxymethyl furfural exists in millet-fried Codonopsis pilosula and rice-fried Codonopsis pilosula.3.Pharmacodynamic research of Codonopsis pilosula participating in different rice frying in rats with spleen deficiencyEach administration group can increase the body mass of rats with spleen deficiency,improve gastrointestinal function,immune function,and water and fluid metabolism indexes,and have certain therapeutic effects on spleen deficiency.Compared with the model group,the last swim time,CK content,the D-xylose excretion rate,small intestine propulsion rate,GAS,MTL,VIP,spleen index,thymus index,total number of white blood cells,AQP 3 and Na~+-K~+-ATPase content in different rice-fried Codonopsis pilosula groups are different Increased;SS,Ig G,TNF-αcontent and CD4~+/CD8~+ratio decreased to varying degrees,indicating that different rice-fried Codonopsis pilosula can improve the symptoms of spleen deficiency in rats to varying degrees.In terms of regulating gastrointestinal function,compared with the millet-fried Codonopsis pilosula group,the GAS content of the rice-fried Codonopsis pilosula group was higher,and there was a significant difference(P<0.05);the two groups had a significant effect on body weight gain,small intestinal advancement rate and urine Although there is no significant difference in the comparison of xylose excretion rate,MTL,VIP,and SS,the rats in the millet-fried Codonopsis pilosula group gained more weight,with higher MTL and VIP content;while the small intestinal propulsion rate and urine D-xylose in the rice-fried Codonopsis pilosula group The excretion rate is higher and the SS content is lower.In terms of immune function regulation,compared with the millet-fried Codonopsis pilosula group,the rice-fried Codonopsis pilosula group had higher thymus index,lower IL-6 content(P<0.05),lower spleen index and TNF-αcontent,total number of white blood cells,CD4~+/The CD8~+ratio and Ig G content were higher,but there was no significant difference(P>0.05).In terms of regulating water metabolism,compared with the millet-fried Codonopsis pilosula group,the Na~+-K~+-ATPase content of the rice-fried Codonopsis pilosula group was lower(P<0.05),and the content of AQP 3 was higher,but there was no significant difference(P>0.05).4.Analysis of the difference of intestinal flora between Codonopsis pilosula and different rice-fried Codonopsis pilosulaA total of 1,800,324 valid sequences and 48,232 OTUs were obtained from 30samples in 5 combinations.The results of principal component analysis(PCA),principal coordinate analysis(PCo A)and non-metric multidimensional scaling analysis(NMDS)based onβdiversity distance show the intestinal flora of rats in the blank group and the model group,the model group and the three administration groups There are obvious differences in composition;at the phylum level,the dominant flora of the 5 groups are Firmicutes,and the secondary dominant flora is Bacteroides;at the genus level,the dominant flora of the 5 groups are Lactobacillus,The secondary dominant flora is the Mycobacterium spp.There is no significant difference in the microbial structure and relative abundance between the millet-fried Codonopsis pilosula group and the rice-fried Codonopsis pilosula group.Conclusion:There is a certain difference between the main chemical composition content of Codonopsis pilosula and the overall chemical composition shown by the fingerprint spectrum of fried rice.Stir-fried Codonopsis pilosula with large and small rice has a certain improvement effect on rats with spleen deficiency,but both have their own advantages in regulating indicators.Among them,fried Codonopsis pilosula with rice is more helpful in regulating the function of the digestive tract and maintaining the integrity of the digestive tract structure.It helps to enhance the immune function of rats with spleen deficiency;the millet-fried Codonopsis pilosula is more helpful to regulate the water metabolism of spleen-deficiency rats.At the same time,Codonopsis pilosula and rice-fried Codonopsis pilosula can regulate the intestinal flora of spleen-deficiency rats.