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Article in Chinese | WPRIM | ID: wpr-887968


As an effective antipyretic medicine,Indigo Naturalis has a long history of application in the field of Chinese medicine.The content of organics,mainly indigo and indirubin,is about 10%. However,the active ingredients and mechanism of its antipyretic effect have not yet been fully elucidated. In view of this,they were investigated in this study with the rectal temperature change as an indicator and 2,4-dinitrophenol-induced fever rats as subjects. The content of PGE2 and c AMP in the hypothalamus and the serum levels of TNF-α,IL-1β and IL-6 were determined by ELISA. Moreover,the plasma samples of fever rats were analyzed by metabonomics in combination with UPLC-Q-TOF-MS for the exploration of potential biomarkers and the discussion on the antipyretic mechanism of Indigo Naturalis and its active ingredients. The results showed that the rising trend of rectal temperature in rats was suppressed 0. 5 h after the treatment with Indigo Naturalis,organic matter,indigo or indirubin as compared with the rats of model group( P < 0. 05),among which Indigo Naturalis and organic matter had better antipyretic effect. ELISA results showed that organic matter and indigo can inhibit the expression of PGE2 and c AMP( P<0. 01),while Indigo Naturalis and organic matter were effective in curbing the increase in TNF-α( P<0. 05). A total of 21 endogenous metabolites were identified from the plasma samples of the Indigo Naturalis,organic matter,indigo and indirubin groups,which were mainly involved in glycerophospholipid metabolism.

2,4-Dinitrophenol , Animals , Antipyretics , Drugs, Chinese Herbal , Indigo Carmine , Indigofera , Rats
Article in Chinese | WPRIM | ID: wpr-887965


The soaking and fermentation of Baphicacanthus cusia( Nees),the important intermediate link of Indigo Naturalis processing,facilitates the synthesis of indigo and indirubin precursors and the dissolution of endogenous enzymes and other effective components,while the role of microorganisms in the fermentation is ignored. The present study investigated the changes of microbial community structure in Indigo Naturalis processing based on 16 S amplicon sequencing and bioinformatics. Meanwhile,the contents of indigo,indirubin,isatin,tryptanthrin,indole glycoside,etc. were determined to explore the correlation between the microorganisms and the alterations of the main components. As demonstrated by the results,the microbial diversity decreased gradually with the fermentation,which bottomed out after the addition of lime. Proteobacteria,Bacteroidetes,and Firmicutes were the main dominant communities in the fermentation. The relative abundance of Proteobacteria declined gradually with the prolongation of fermentation time,and to the lowest level after the addition of lime. The relative abundance of Firmicutes increased,and that of Bacteroidetes decreased first and then increased. The contents of effective substances in Indigo Naturalis also showed different variation tendencies. As fermentation went on,indole glycoside decreased gradually; indigo first increased and then decreased; indirubin and isatin first decreased and then increased; tryptanthrin gradually increased. Those changes were presumedly related to the roles of microorganisms in the synthesis of different components. This study preliminarily clarified the important role of microorganisms in the soaking and fermentation and provided a scientific basis for the control of Indigo Naturalis processing and the preparation of high-quality Indigo Naturalis.

Fermentation , Indigo Carmine , Indigofera , Indoles , Microbiota
Article in Chinese | WPRIM | ID: wpr-690505


In order to explore appropriate measures to promote germination after the harvest of Epimedium pseudowushanense, 6-BA, urea, ammonium bicarbonate and GA₃ were chosen to spray on the root and rhizomes, and then the biological indicators such as branches, leaf length, leaf width, plant height and so on, were measured in different periods, and the contents of epimedin A, epimedin B, epimedin C and icariin in the dry leaves were detected by HPLC. Results showed that 6-BA 90 mg·L⁻¹(B1), 6-BA 60 mg·L⁻¹(B2),6-BA 30 mg·L⁻¹+urea 300 mg·L⁻¹ (C1), 6-BA 60 mg·L⁻¹+urea 300 mg·L⁻¹(C2),6-BA 60 mg·L⁻¹+ ammonium bicarbonate 300 mg·L⁻¹(C4) significantly increased bud germination in the early period, and the plants quickly set up new system of photosynthesis, the branches in a month of which were higher than the control group respectively by 165.9%, 115.76%, 103.86%, 104.50%, 81.67%.However the branches developed the next year and the dry weight of leaves per plant in group B1 and B2 were much lower than that in control group. The groups that use 6-BA and nitrogen at the same time reaped a good yield of leaves even though the treatment had no significant influence on the branches developed the next year. The dry weight of leaves of C1, C2, C4 treatments were 36.80%, 32.84%, 45.97% more than the control group respectively. Therefore, C1, C2 and C4 treatments are the more appropriate to promote recovery after harvest. Furthermore, different groups, except 10 mg·L⁻¹ 6-BA treatment significantly reduced the content of epimedin C, other groups didn't have any significant effect on the contents of such flavonoids.

Article in Chinese | WPRIM | ID: wpr-335889


In this study, 128 individuals form 14 Epimedium pubescens populations and 1 E. stellulatum population were analyzed by ISSR marker. The data were calculated by POPGENE software and clustered by UPGMA method. Optical microscope was used to observe the main types of the non-glandular hairs and their characteristics in each population. It is found that the following conclusions: Non-glandular hairs can be divided into five morphological categories, long straight pubescent, curly pubescent, appressed curly pubescent, pseudo short appressed hairs and long appressed. Eight primers were screening and a total of 94 bands were detected in ISSR, among which 90 were polymorphic bands. Based on the results of ISSR cluster analysis, 15 populations were divided into 3 clades. E. stellulatum populations should be incorporated into the E. pubescens or as avariety under E. pubescens not be independent and as it has no separate phylogenetic branch for a cluster. The genetic relationship among the populations of E. pubescens was closely related with its geographical distribution and non-glandular hair features. But there were also some inconsistency, which provided a good hint for the further study on the interspecific relationship and natural speciation manner of Epimedium species. Population diversity analysis showed Nm=0.354 4, Nei's=0.585 2. It was showed that E. pubescens has high genetic diversity among populations, for which the main reason was probably the high inbreeding rate and the small range of seed dispersal.

Article in Chinese | WPRIM | ID: wpr-338235


Nitrogen nutrition divided into concentrations and forms for Epimedium pseudowushanense was studied under hydroponic conditions. There were 7 nitrogen concentrations treatments(N0-N6)which nitrogen concentration was 0, 2.5, 5.0, 7.5, 10.0, 13.0 mmol•L⁻¹, respectively, and 5 nitrogen forms and proportions treatments (T1-T5) which NO3-N/NH4-N was 5∶0, 4∶1, 2.5∶2.5, 1∶4, 0∶5 respectively, the total nitrogen applied was 5 mmol•L⁻¹. The results showed that along with the increase of nitrogen concentration, biomass (dry weight) and leaf dry weight increased, which in N5 treatment was 29%, 23% higher than those in N0 and 36%, 23% higher than those in N4 respectively, but not significantly different from N6 treatment. Roots and stem dry weights did not change significantly. Pn and the contents of photosynthetic pigments were higher in N3-N5 treatments. Epimedin A, B, C, icariin and total flavonoids contents were the lowest in N3 treatment, suddenly increase in N4 treatment and only a little change between N4-N6 treatments. N5 treatment with the highest epimedin C content was 131% higher than N3 treatment with the lowest epimedin C content (P<0.05). While in the nitrogen forms and proportions treatments, biomass, leaf and stem dry weights were the highest in T3 treatment and lower in T1 and T5 treatments, without significance between T1 and T5 treatments. These indicators in T3 treatment were more than 30% higher than those in T1 treatment. The impact of nitrogen concentrations was greater than nitrogen forms on flavonoids. epmedin C content was the highest in T1 treatment, only 5%-8% higher than that in T2-T5 treatments (no significance between 4 treatments). Epimedin A, B, icariin and total flavonoids contents were higher in T1 treatment, the lowest in T2 treatment, those in T1 treatment was 41%, 62% and 27% higher than T2 treatment respectively (P<0.05). In a conclusion, E. pseudowushanense is a high nitrogen tolerant plant, 10.0 mmol•L⁻¹ was the boundary point between growing general to well and the optimal nitrogen concentration for it was 10.0-13.0 mmol•L⁻¹. Besides, it prefers to grow in NO-3 plus NH+4 and the optimal proportion was 2.5∶2.5.

Article in Chinese | WPRIM | ID: wpr-250425


Epimedium is a widely used medicinal plants. Due to excessive use of wild resources and resource constraints, artificial cultivation is the only way to achieve sustainable use of resources. E. pseudowushannense is intended to achieve the first cultivated species, but plant production technology is not yet fully mature.The diurnal variation of photosynthetic and eco-physiological characteristics of E. pseudowushannense in different growing age was studied by the CI-340 Portable Photosynthesis System under natural conditions. The net photosynthetic rate (Pn), stomatal conductance (C), intercellular CO2 concentration (Ci), photosynthetically active radiation (PAR), relative humidity (RH), air temperature (Ta), apparent mesophyll conductance (ALMC), transpiration rate (E), leaf temperature (Tl), water use efficiency (WUE) and other photosynthetic physiological factors and environmental factors were measured. The obtained data was analyzed by correlation analysis, path analysis and stepwise multiple-regression analysis to explore the relationship between net photosynthetic rate and physiological and ecological factors. The results show that: ①The daily variation of the Pn, E, ALMC, C of E. pseudowushannense in different growing age took on"two peak type", the Pn was with an obvious midday depression phenomenon at noon.②The Pn, ALMC and C of annuals' were significantly higher than biennial or three-year; but the Ci was basically a three-year > biennial> annual.③The correlation analysis, path analysis and stepwise regression analysis showed that ALMC, Ci, Ta might played important role in the Pn and ranked as ALMC>Ci>Ta.