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ObjectiveTo clarify the differences in the efficacy and mechanism of different processed products of Atractylodes chinensis rhizoma by the pharmacodynamics and metabolomics studies of raw， bran-fried and rice water-processed products on rats with spleen deficiency. MethodSixty male SD rats were randomly divided into blank group， model group， raw product group（3.75 g·kg-1）， bran-fried product group（3.75 g·kg-1）， rice water-processed product group（3.75 g·kg-1） and Shenling Baizhusan group（6.7 g·kg-1）， with 10 rats in each group. The method of excessive fatigue+improper diet was used to establish a spleen deficiency model in rats. After the end of modeling， except for the blank and model groups， each dosing group was given the corresponding drug suspension， the immune organ coefficients of each group of rats were examined， the levels of interleukin-6（IL-6）， tumor necrosis factor-α（TNF-α）， immunoglobulin G（IgG）， amylase（AMS）， motilin（MTL）， gastrin（GAS）， Na+-K+-adenosine triphosphatase（ATPase）， aquaporin 2（AQP2）， AQP3 and AQP8 in rats were measured by enzyme-linked immunosorbent assay（ELISA）. Ultra high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS） combined with orthogonal partial least squares-discriminant analysis（OPLS-DA） were used to search for biomarkers in the plasma samples of spleen-deficient rats by using two criteria［P<0.05 and variable importance in the projection（VIP） value>1］， and to compare the different modulatory effects of the three decoction pieces on the splenic-deficient biomarkers， and metabolic pathway analysis was conducted through the Kyoto Encyclopedia of Genes and Genomes（KEGG） database. ResultCompared with the blank group， the thymus index and spleen index of rats in the model group were significantly decreased（P<0.05）， the levels of IL-6， TNF-α， IgG and AQP2 were significantly increased（P<0.05）， the levels of AMS， GAS， MTL， AQP3， AQP8 and Na+-K+-ATPase were significantly decreased（P<0.05）. Compared with the model group， raw products， bran-fried products and rice water-processed products all increased thymus index and spleen index（P<0.05）， decreased IL-6， TNF-α， IgG and AQP2 levels（P<0.05）， and increased AMS， GAS， MTL， AQP3， AQP8 and Na+-K+-ATPase levels to different degrees. A total of 176 differential metabolites were screened in the model group compared with the blank group， of which 75， 72 and 84 biomarkers were called back by the raw products， bran-fried products and rice water-processed products， respectively（P<0.05， P<0.01）. Raw products of A. chinensis rhizoma mainly affected glycine， serine and threonine metabolism. Bran-fried products mainly affected alanine， aspartate and glutamate metabolism， D-arginine and D-ornithine metabolism. Rice water-processed products mainly affected glycine， serine and threonine metabolism， alanine， aspartate and glutamate metabolism， citrate cycle， thiamine metabolism， D-arginine and D-ornithine metabolism. ConclusionRaw products， bran-fried products and rice water-processed products of A. chinensis rhizoma all have good spleen strengthening effects， among which the effects of bran-fried products and rice water-processed products were stronger. Meanwhile， raw products has the strongest dryness， followed by bran-fried products， and the weakest dryness of rice water-processed products. The three decoction pieces are able to significantly modulate metabolic abnormalities in spleen-deficient rats， and the mechanism may be related to amino acid metabolism such as glycine， serine and threonine metabolism as well as alanine， aspartate and glutamate metabolism.
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Objective:To use rbcL sequences to identify the rhizomes of the Liaoning collection of Atractylodes chinensis (DC.) Koidz.; To provide a basis for ensuring the feasibility of cultivation of the native herb in Liaoning Province. Methods:A total of 30 rhizomes of Atractylodes chinensis (DC.) Koidz. were collected from 10 regions cultivated in Liaoning Province, and the total DNA was extracted. DNA barcodes were screened by PCR, and the rbcL sequences of the samples were amplified and sequenced, and the amplification and sequencing success rates were calculated. Sequence alignment was performed using MEGA 7.0 software; a systematic clustering tree was constructed using the neighbour-joining method. Results:The success rates of DNA extraction from the rhizomes of Atractylodes chinensis (DC.) Koidz. were all 93.3%, and the success rates of PCR amplification and sequencing were all 100%. Among the 30 samples of Atractylodes chinensis (DC.) Koidz. in Liaoning Province, two samples had intraspecific variation, and the rest of the base sequences of Atractylodes chinensis (DC.) Koidz. were identical. Atractylodes chinensis (DC.) Koidz. was closer to the herbs of the genus Cangzhu, a relative species of Asteraceae, and was genetically more distant from the rest of Asteraceae. The NJ tree could distinguish Atractylodes chinensis (DC.) Koidz. and its relatives. Conclusion:The quality of Atractylodes chinensis (DC.) Koidz. cultivars in Liaoning Province is basically similar, and the rbcL sequence can be used as a valid sequence fragment for the identification of Atractylodes chinensis (DC.) Koidz. DNA barcode.
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Aim To analyze the differences in plasma biomarkers and metabolic pathways between Atractylodes chinensis and Atractylodes coreana after intervention in spleen deficiency rats, and discuss the spleen strengthening mechanism of the two from a non targeted metabolomics perspective. Methods A spleen deficiency model was established in SD rats using a composite factor method of improper diet, excessive fatigue, and bitter cold diarrhea. To determine the content of gastrointestinal and immunological indicators, UHPLC-QE-MS technology was used, combined with principal component analysis (PC A) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) methods to search for biomarkers in plasma of spleen deficiency rats, and metabolic pathways were induced using the Pathway database. Results After administration of Atractylodes chinensis and Atractylodes coreana, various indicators in plasma of spleen deficiency rats showed varying degrees of regression. Metabolomics analysis showed that Atractylodes chinensis and Atractylodes coreana respectively recalled 70 and 82 plasma differential metabolites. Atractylodes chinensis mainly regulated two metabolic pathways : "Glycine, serine, and threonine metabolism, and "Thiamine metabolism". Atractylodes coreana mainly regulated five metabolic pathways, "Glycine, serine, and threonine metabolism", "Thiamine metabolism, "Pyrimidine metabolism", "Butanoate metabolism", and "Riboflavin metabolism". Conclusions Both Atractylodes chinensis and Atractylodes coreana have certain regulatory effects on spleen deficiency rats, and their mechanism of action may be related to regulating metabolic pathways such as "Glycine, serine, and threonine metabolism, and "Thiamine metabolism"in spleen deficiency.
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ObjectiveTo explore the biological mechanism of drought improving the quality of Rhizoma Atractylodis Chinensis and establish a new method for the production of high-quality medicinal materials. MethodThe fresh roots of Atractylodes chinensis were soaked in 0 （control）， 5%， 10%， and 20% PEG-6000 solutions. The changes in reactive oxygen species （ROS） level， antioxidant enzyme activity， activities of key enzymes in primary metabolism and secondary metabolism， and content of secondary metabolites were compared. ResultCompared with the control group， the treatment with 20% PEG for 2 days elevated the levels of superoxide anion radicals （O2-·）， hydrogen peroxide （H2O2）， and malondialdehyde （MDA） by 172.5%， 56.9%， and 14.7%， respectively. The treatment did not change the activity of superoxide dismutase （SOD）， reduced the peroxidase （POD） activity， and increased the catalase （CAT） activity by 10.8%. It increased the activities of 3-hydroxy-3-methylglutaryl-CoA reductase （HMGR）， phosphoenolpyruvate carboxylase （PEPC）， and acetyl-CoA carboxylase （ACC） by 49.9%， 12.1%， and 19.0%， respectively. Furthermore， the content of atractylodin， β-eudesmol， atractylone， and atractylenolide Ⅱ was increased by 51.0%， 36.9%， 47.1%， and 91.5%， respectively. The simulated drought stress can cause the burst of ROS in the fresh roots of A. chinensis， induce the physiological state of plants under drought， change the antioxidant system， and promote the massive synthesis of secondary metabolites in a short time. ConclusionPEG-6000-simulated drought stress can greatly improve the quality of A. chinensis in cultivation.
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italic>Atractylodes chinensis has important medicinal and economic values. In this study, the chloroplast genome sequences of four A. chinensis samples from different producing areas were sequenced using the Illumina platform. The specific DNA barcodes were screened and the germplasm resources of A. chinensis samples from different producing areas and the genetic diversity of the population were analyzed basing on the specific barcodes. The whole chloroplast genomes of the four A. chinensis samples had a typical cyclic tetrad structure, with 112 genes annotated. The comparative genomics results indicated that ccsA and trnC-GCA_petN were potential specific DNA barcodes for intraspecific identification of A. chinensis. Polymerase chain reaction (PCR) analysis of ccsA and trnC-GCA_petN was performed on 256 samples from 14 areas in 9 provinces, and the amplification efficiency was 100%. Sequence analysis showed that ccsA and trnC-GCA_petN had 11 and 22 variant positions, which could identify 16 and 22 haplotypes, respectively. The combined sequence analysis identified 39 haplotypes, named Hap1-Hap39, of which the most abundant and widely distributed genotype was Hap9. Haplotype diversity (Hd) = 0.896 and nucleotide diversity (Pi) = 0.002 22 indicated high genetic diversity at the species level in A. chinensis. The genetic distances of the haplotypes were 0.000 00-0.004 88, indicating that there were small genetic differences among the haplotypes. The results of phylogenetic tree analysis showed that 39 haplotypes had very close genetic relationship, and formed two obvious branches with other groups of the same genus except Atractylodes macrocephala. This study plays an important role in the identification of the origin of A. chinensis and the protection and breeding of germplasm resources.
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ObjectiveTo investigate the effect of exogenous H2O2 on secondary metabolism in Atractylodes chinensis and its mechanism. MethodFresh rhizomes of A. chinensis were treated with 5.0， 1.0， 0.2， 0.04 mmol·L-1 H2O2 solution and clean water， and the relationships between the contents of reactive oxygen species， activities of antioxidant enzymes， activities of key enzymes of secondary metabolites， and contents of secondary metabolites in A. chinensis were compared. ResultUnder treatment with exogenous H2O2， the content of reactive oxygen species and malondialdehyde （MDA） in the fresh rhizomes of A. chinensis were significantly elevated on the 4th day， and returned to normal level on the 6th-8th day. The activities of superoxide dismutase （SOD）， catalase （CAT） and peroxidase （POD） were all increased first and then decreased， and reached the peak on the 4th， 4th-6th and 2th-4th day， respectively. The activities of 3-hydroxy-3-methyl glutaryl coenzyme A reductase （HMGR） and acetyl CoA carboxylase （ACC）， key enzymes of the secondary metabolites， were remarkably enhanced， and under treatments with different concentrations of H2O2， the activities of key synthetic enzymes of the secondary metabolites in 0.2 mmol·L-1 H2O2 group were increased most， with the highest biosynthesis of secondary metabolites. The contents of atractylodin， β-eudesmol， atractylone， atractylenolide Ⅱ， and atractylenolide Ⅲ on the 6th day of 0.2 mmol·L-1 H2O2 treatment were 89.5%， 108.7%， 308.8%， 64.7% and 9.3%， respectively higher than those in the control. ConclusionThe antioxidant enzymes and secondary metabolites in A. chinensis synergistically maintain the balance of reactive oxygen species， and exogenous H2O2 can improve the medicinal quality of A. chinensis remarkably.
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We explored the correlations between the color difference values [ΔL~*(lightness), Δa~*(red-green), Δb~*(yellow-blue)] and the content of four active components(including sesquiterpenoids and polyacetylenes) in the powder of Atractylodes lancea and A. chinensis, aiming to provide reference for the quality evaluation of Atractylodis Rhizoma and establish a qualitative model that can distinguish between A. lancea and A. chinensis based on the chromatic values. The tristimulus values(L~*, a~*, and b~*) of 23 batches of A. lancea and A. chinensis were measured by a color difference meter. The content of atractylenolide Ⅱ, β-eudesmol, atractylodin, and atractylone in the 23 batches of samples were measured by high performance liquid chromatography(HPLC). Principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) were performed to establish the qualitative models for distinguishing between A. lancea and A. chinensis. SPSS was employed to analyze the correlations between the tristimulus values and the content of the four index components. The results showed that the established PCA and PLS-DA models can divide the A. lancea and A. chinensis samples into two regions, and the tristimulus values of A. lancea and A. chinensis were positively correlated with the content of β-eudesmol and atractylodin. Therefore, the PCA and PLS-DA models can successfully identify A. lancea and A. chinensis, and the appearance color can be used to quickly predict the internal quality of Atractylodis Rhizoma. This study provides a reference for the quality evaluation of Atractylodis Rhizoma and the modern research on the color of Chinese medicinal materials.
Subject(s)
Atractylodes , Sesquiterpenes, Eudesmane , Drugs, Chinese Herbal , Rhizome , ExcipientsABSTRACT
Objective:The effects of Atractylodes lancea， A. coreana， A. japonica， A. chinensis and Atractylodis Macrocephalae Rhizoma on spleen-Qi deficiency rats were compared. Method:A model of spleen-Qi deficiency was induced in rats by diet and overwork.The rats are given different suspensions of A.japonica， A.chinesis， A.coreana， A.lancea and Atractylodis Macrocephalae Rhizoma To test the indicators of the digestive system， immune system and antioxidant enzyme system related to spleen deficiency.Compare the similarities and differences between Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from four different sources. Result:All the drug-administered groups can increase the levels of γ interferon （IFN-γ）， gastric secrete element（GAS），serum amylase （AMS） and catalase（CAT） in rats with spleen-deficiency syndrome，in addition to the CAT index， the other indicators of the A. coreana， A.japonica， and A.lancea were significantly increased（P<0.05）. The MTL content of the A.chinesis， A.lancea， A.coreana and A.japonica increased and was significant（P<0.05）. The SDH content of A.japonica.and A.chinesis increased， and the difference was not significant.The increase of GSH-Px in the A.chinesis is significant（P<0.05）. All the drug-administered groups can reduce the content of IgG， TNF-α and MDA in rats with spleen deficiency and deficiency syndrome. Among them， the IgG content of the A.chinesis. and the A.lancea was significantly decreased（P<0.05）. The content of TNF-α in A.japonica group was significantly decreased（P<0.05）. The content of MDA in the A.chinesis， the A.lancea， the A.coreana，the A.japonica and the Atractylodis Macrocephalae Rhizoma were significantly decreased（P<0.05）.The decrease of IL-6 in the A.lancea was significant（P<0.05）. Conclusion:Four different sources of Atractylodes Rhizome and A.macrocephala have certain therapeutic effects on spleen-deficiency rats with deficiency syndrome.The therapeutic effect of A.lancea and A.japonica is basically the same，regulating the absorption，secretion and elimination of inflammation in the digestive system of rats with spleen deficiency A.coreana， A.chinesis， and Atractylodis Macrocephalae Rhizoma have certain regulatory effects in the digestive system， digestive tract inflammation， and antioxidant enzymes.
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OBJECTIVE：To simultaneo usly determine the contents of atractylenolide Ⅱ ，β-eudesmol，atractyloxin and atractylone in Atractylodes chinensis ，and to evaluate the quality of A. chinensis with different growth years combined with color difference principle. METHODS ：HPLC method was adopted. The determination performed on Agilent Eclipse XDB-C 18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid （gradient elution ）at the flow rate of 1.0 mL/min；the detection wavelengths were set as 208 nm（atractylenolide Ⅱ，β-eudesmol），340 nm（atractyloxin）and 220 nm（atractylone）；the sample size was 15 μ L. Using atractyloxin as reference，QAMS was adopted to establish relative correction factors （RCFs） of atractylenolideⅡ，β-eudesmol and atractylone ；the content of each component in A. chinensis with different growth years were calculated. The contents of above 4 components were determined by external standard method and then compared with the results of QAMS. The color difference values of A. chinensis powder were measured based on color difference principle. The correlation analysis of above 4 components content with color was carried out by Pearson correlation analysis. RESULTS ：The separation degree of atractylenolide Ⅱ，β-eudesmol，atractyloxin and atractylone in A. chinensis was higher than 1.5. The linear range were 1.01-10.10，3.30-33.00，4.40-44.00，5.34-53.40 μg/mL，respectively. RSDs of precision ，reproducibility and stability tests were all lower than 2％，while the average recovery rates were 101.34％-104.67％（RSD＜1.5％，n＝6）. Using atractyloxin as reference ， RCFs of atractylenolide Ⅱ，β-eudesmol and atractylone were 3.896 7，5.928 2，9.727 9，with RSD of 0.35％，2.89％，0.36％ （n＝6），respectively. Relative deviation of 3 components （except for atractyloxin ） in 24 batches of A. chinensis ranged 0.03％-1.45％ between QAMS and external standard method ，which indicated that the results of two methods were consistent ，and the content of each component increased with the increase of growth years. Atractylenolide Ⅱ，β-eudesmol，atractyloxin and atractylone in A. chinensis had significant negative correlation with its color shade （L*），total color difference （E*ab）（P＜0.01）， and significant positive correlation with color red-green direction （a*）， color yellow-blue direction （b*）（P＜0.01）. CONCLUSIONS：The established HPLC-QAMS method can be used for the determination of atractylenolide Ⅱ，β-eudesmol， atractyloxin and atractylone in A. chinensis . The longer the growth period is ，the higher each component content is. The color of A. chinensis is closely related to the content of each component ，and the content of effective components is higher in A. chinensis with dark yellowish brown color.
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OBJECTIVE To observe the preventive effect on nosocomial infection by the herbal medicine Atractylodes chinensis disinfectant for air sterilization in cardiothoracic surgery.METHODS A.chinensis disinfectant and ultraviolet irradiation were used to disinfect the patients′ rooms of our cardiothoracic surgery department.The total number of germ and fungus in the indoor air before and after using the two methods were collected.The nosocomial infection rates of the same period were also studied.RESULTS A.chinensis disinfectant had strong effect on sterilization of indoor germs and fungi,and the sterilization rate was 92.7%.The comparison between A.chinensis disinfectant and ultraviolet irradiation showed significant difference in their disinfection effects,while no significant difference in their nosocomial infection rates.CONCLUSIONS The herbal medicine A.chinensis disinfectant can achieve good sterilization effect and prevent nosocomial infection.