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Objective: To investigate the effect and the mechanism of Astragalus membranaceus (Huangqi in Chinese, HQ) extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii (Fuzi in Chinese, FZ) in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier. Methods: Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d. Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids. Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins (TJ, including Claudin-1, Occludin and ZO-1) were measured using Western blot and real-time PCR, respectively. The location and expression of TJ protein was also investigated by the immunofluorescence method. Results: Compared with the normal group, the protein expression of MRP2, BCRP and TJ proteins in the model group were significantly down-regulated. After oral administration of HQ, the alkaloid absorption in intestinal villi was inhibited, MRP2, BCRP and TJ proteins were up-regulated, the green fluorescence staining of Claudin-1, Occludin, and ZO-1 was enhanced, and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity. Conclusion: HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components, leading to a decrease in the absorption of drug-like molecules.
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In Hezhang county, Guizhou province, black spot tends to occur to Aconitum carmichaelii in the hot rainy summer, with the incidence up to 50%-70%, seriously impacting the yield and quality of the medicinal material. Thus, this study aims to clarify the pathogen and the occurrence characteristics. To be specific, the pathogen was isolated and identified according to Koch's postulates and the pathogenicity and biological characteristics were determined. In addition, the sensitivity of the pathogen to four microbial fungicides, four botanical fungicides, and five chemical fungicides was determined with the mycelium growth rate method for the purpose of screening out optimal fungicides. The pathogen was identified as Alternaria alternate, as evidenced by the similar colony morphology and microscopic characteristics and 99.55%-100% similarity in sequences of rDNA-ITS, LSU, 18S, and TEF of the two. The optimum growth conditions for A. alternata were 28 ℃, pH 8, and continuous darkness. Bacillus subtilis had strong inhibitory effect on the pathogen, and the inhibition rate was more than 90% when the concentration was 1 mg·L~(-1). In addition, difenoconazole and quinoline copper can also control the pathogen, with median effective concentration(EC_(50)) of 2.92 and 9.02 mg·L~(-1), respectively. This study lays a theoretical basis for the field control of black spot in A. carmichaelii.
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Aconitum , Alternaria , Fungicides, Industrial/pharmacology , MyceliumABSTRACT
Aconiti Lateralis Radix Praeparata (Fuzi in Chinese) is a commonly used Chinese materia medica in clinic, with the effects in rescuing from collapse by restoring yang, eliminating cold to stop pain, warming yang and transforming qi. C19 diterpenoid alkaloids from Fuzi can be divided into three types: diester type, monoester type and amine alcohol type, which can lead to neurotoxicity and cardiotoxicity while exerting anti-inflammatory, analgesic, anti-tumor and other pharmacodynamic effects. In this paper, the chemical structure, pharmacology and toxicological effects of C19 diterpene alkaloids of Fuzi were systematically combed, in order to provide a theoretical basis for safer and more effective use of Fuzi in clinic.
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Objective: To study the chemical constituents in the aerial parts of Aconitum carmichaelii. Methods: The air-dried arial parts of A. carmichaelii were powdered and extracted with methanol by percolation extraction. After the removal of solvent under reduced pressure, the crude extract was dissolved in 1.5% HCl solution, and then extracted by ethyl acetate to obtain the total crude extract. The compounds were isolated and purified by column chromatography and identified by spectral analyses (MS, 13C-NMR, and 1H-NMR). Results: Fifteen compounds were isolated from A. carmichaelii and characterized as indol-3-carboxylic acid (1), corchoionol C (2), β-sitosterol-3-O-β-D-glucoside-6’-palmitate (3), (+)-pinoresinol (4), (+)-N-formylnorglaucine (5), oxoglaucidaline (6), glaucine (7), (+)-cataline (8), kaempferol-7-O-α-L-rhamnofuranoside (9), kaempferol-3-O-β-(2″-acetyl)-galactopyranoside (10), megastigmane (11), kaempferol-7-O-α-L-arabinoside (12), kaempferol-3-O-β-D-xylopyranoside (13), kaempferol-3-O-β-D- glucopyranoside (14), and quercetin-3-O-β-D-galactopyranoside (15). Conclusion: All compounds are isolated from aerial parts of A. carmichaelii for the first time, and compounds 1-3,5-6,8-15 are isolated from this plant for the first time.
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Objective: To observe the effects of Aconitum carmichaelii on serum metabolites in mice by metabolomics technology, and to explore biomarkers and metabolic pathways and targets related to its treatment of various ailments such as cardiovascular diseases, in order to uncover its molecular mechanism of efficacy. Methods: Twenty male mice were randomly divided into two groups, and A. carmichaelii decoction and distilled water were orally administered with the dose of 15 mL/(kg∙d) for consecutive 4 d respectively. Collected blood samples of each group were analyzed using UPLC-MS/MS technology, and data pattern recognition was performed using PCA, PLS-DA and other analytical methods. Meanwhile, differential metabolites were screened out based on VIP greater than 1 and manual integral calculation. The differential metabolites were used for pathway analysis. Network modular analysis and targets screening were performed by Cytoscape and MetScape. Results: When performing data pattern recognition, the aconite group and the control group could be completely separated. A total of 18 differential metabolites were screened out, and their contents were up-regulated. Pathway analysis was performed to obtain five related pathways, namely linoleic acid metabolism, arachidonic acid metabolism, starch and sucrose metabolism, nicotinate and nicotinamide metabolism, and inositol phosphate metabolism. Fourteen modules were obtained using the network analysis, the largest two of which were arachidonic acid metabolism pathway and linoleic acid metabolism pathway. The degree of arachidonic acid (59), linoleic acid (55), nicotinamide (26), and palmitic acid (11) were greater than the mean value (8.010) in the network, and the related pathways were arachidonic acid metabolism, linoleic acid metabolism, nicotinate and nicotinamide metabolism, and saturated fatty acid beta-oxidation pathway respectively. A total of 26 genes were screened out, all of which belonged to the cytochrome P450 enzyme system. Conclusion: A. carmichaelii may affect arachidonic acid metabolism by acting on CYP450, thereby improving the body's energy metabolism and producing therapeutic effects.
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OBJECTIVE： To study the effects of Aconitum carmichaelii water extract on the expression of 3 kinds of efflux transporters and 3 kinds of tight junction proteins as well as their genes in duodenum tissues of rats. METHODS： Thirty-two SD male rats were randomly divided into normal group， A. carmichaelii water extract low-dose， medium-dose and high-dose groups [0.45， 0.9， 1.8 g/（kg·d）， by crude drug]， with 8 rats in each group. They were given water and relevant liquid 0.1 mL/kg intragastrically for consecutive 7 d. After last administration， the duodenal segments of rats were collected. Western blotting assay was used to detect the expression of efflux transporters as P-glycoprotein （P-gp）， breast cancer resistance protein （Bcrp）， multidrug resistance protein 2 （Mrp2） as well as tight junction proteins as Claudin-1， Occludin and ZO-1. mRNA expression of 6 kinds of proteins relevant gene were determined by qRT-PCR respectively. RESULTS： Compared with normal group， the protein expression of P-gp， Mrp2， Bcrp， Claudin-1， Occludin and ZO-1 in duodenum of rats were increased significantly in A. carmichaelii water extract groups （P＜0.01）. mRNA expression of P-gp in A. carmichaelii water extract groups， mRNA expression of Bcrp in A. carmichaelii water extract low-dose and high-dose groups as well as mRNA expression of Claudin-1 in A. carmichaelli water extract medium-dose and high-dose groups were increased significantly， with statistical significance （P＜0.05 or P＜0.01）. There was no statistical significance in mRNA expression of other genes （P＞0.05）. CONCLUSIONS： A. carmichaelii water extract can up-regulate the expression of 3 kinds of efflux transporters and 3 kinds of tight junction proteins at the level of mRNA and/or protein， thus may interact with other substrates of the aforementioned efflux transporters and drugs with cell bypass pathway as the main transport pathway. In clinical practice， adjustment of dosage may be considered in drug combination.
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OBJECTIVE:To optimize formulation matrix of processed Aconitum carmichaelii hydrogel patch,and to investigate its in vitro drug release.METHODS:The ratio of NP700,dihydroxyaluminum aminoacetate,tartaric acid and PVP K90 in processed A.carmichaelii hydrogel patch matrix was optimized by central composite design-response surface method (CCD-RSM) with initial adhesion,peel strength and sensory evaluation as evaluation indexes.The modified Franz diffusion cell method was used for in vitro drug release test processed A.carmichaelii hydrogel patch using accumulative release amount of six ester type alkaloids benzene [benzoyl mesaconine (BM),benzoyl aconitum (BA),benzoylhy paconine (BH),mesaconitine (MT),hypaconitine (HT) and aconitine (AT)] as evaluation indexes.RESULTS:The optimal matrix formulation was NP700-dihydroxyaluminum aminoacetate-tartaric acid-PVP K90 (1.72 ∶ 0.10 ∶ 0.02 ∶ 1.65,m/m/m/m).In validation test,the contents of six ester type alkaloids were 52.77,28.52,28.78,8.81,8.75,8.21 μg/g(RSD＜5％,n=3),comprehensive score was 118.67 ± 1.33 (RSD=4.62％,n=3).The release behavior of BM in vitro conformed to the Higuchi equation.The release behaviors of other 5 alkaloids were consistent with the Higuchi equation.12 h accumulative release amounts of BM,BA,BH,MT,HT and AT were 12.04,2.95,3.55,2.64,2.48,1.97 μg/cm2,respectively.CONCLUSIONS:The processed A.carmichaelii hydrogei patch prepared by matrix prescription is good in appearance,adhesion and in vitro release.The research can provide a basis for the development of new dosage form of processed A.carmichaelii.
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According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloids in mother roots, fibrous roots, stems, and leaves of Aconitum carmichaelii Debx. were measured by HPLC-MS/MS. And multivariate analysis methods, such as clustering analysis and principal component analysis, were applied to analyze the difference among various parts. In addition, the acute toxicity, analgesia, and anti-inflammatory tests were carried out. The results suggested that the contents of alkaloids in mother roots and fibrous roots were approximate, but those of leaves and stems were different from mother roots and fibrous roots. The results of the acute toxicity testing demonstrated the toxicity of fibrous root was strongest, and mother roots were slightly less toxic than fibrous roots. The stems and leaves were far less toxic than mother and fibrous roots. In addition, the analgesia and inflammatory tests showed the effects of the various tissues had no difference each other. These results provided a basis for developing new complementary and alternative treatments for rheumatoid arthritis patients. Simultaneously, the approach may also turn wastes into treasure and promote the development of circular economy.
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Animals , Female , Humans , Male , Mice , Aconitum , Chemistry , Alkaloids , Chemistry , Toxicity , Anti-Inflammatory Agents , Chemistry , Toxicity , Arthritis, Rheumatoid , Drug Therapy , Chromatography, High Pressure Liquid , Drug Evaluation, Preclinical , Drugs, Chinese Herbal , Chemistry , Toxicity , Plant Leaves , Chemistry , Plant Roots , Chemistry , Plant Stems , Chemistry , Tandem Mass SpectrometryABSTRACT
According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloids in mother roots, fibrous roots, stems, and leaves of Aconitum carmichaelii Debx. were measured by HPLC-MS/MS. And multivariate analysis methods, such as clustering analysis and principal component analysis, were applied to analyze the difference among various parts. In addition, the acute toxicity, analgesia, and anti-inflammatory tests were carried out. The results suggested that the contents of alkaloids in mother roots and fibrous roots were approximate, but those of leaves and stems were different from mother roots and fibrous roots. The results of the acute toxicity testing demonstrated the toxicity of fibrous root was strongest, and mother roots were slightly less toxic than fibrous roots. The stems and leaves were far less toxic than mother and fibrous roots. In addition, the analgesia and inflammatory tests showed the effects of the various tissues had no difference each other. These results provided a basis for developing new complementary and alternative treatments for rheumatoid arthritis patients. Simultaneously, the approach may also turn wastes into treasure and promote the development of circular economy.
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Animals , Female , Humans , Male , Mice , Aconitum , Chemistry , Alkaloids , Chemistry , Toxicity , Anti-Inflammatory Agents , Chemistry , Toxicity , Arthritis, Rheumatoid , Drug Therapy , Chromatography, High Pressure Liquid , Drug Evaluation, Preclinical , Drugs, Chinese Herbal , Chemistry , Toxicity , Plant Leaves , Chemistry , Plant Roots , Chemistry , Plant Stems , Chemistry , Tandem Mass SpectrometryABSTRACT
Eight new C-diterpenoid alkaloid arabinosides, named aconicarmichosides E-L (-), were isolated from an aqueous extract of the lateral roots of (Fu Zi). Their structures were determined by spectroscopic and chemical methods including 2D NMR experiments and acid hydrolysis. Compounds -, together with the previously reported four neoline 14--arabinosides from the same plant, represent the only examples of glycosidic diterpenoid alkaloids so far. At a dose of 1.0 mg/kg (i.p.), as compared with the black control, compounds , , and - exhibited analgesic effects with >65.6% inhibitions against acetic acid-induced writhing of mice. Structure-activity relationship was also discussed.
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Objective To identify molecular markers associated with weight of fleshy root of Aconitum carmichaelii, and provide technologic basis to A. carmichaelii breeding. Methods The marker-trait association (MTA) analysis for the weight of first and second fleshy root was conducted in association mapping panel comprised 126 diverse lines based on 280 AFLP loci. Results The population structure analysis of all lines conducted using STRUCTURE indicated that the likely number of subgroups was 2 based on the change in k. A total of 19 loci were identified to associate with 0.87%-6.69% of phenotypic variance for objective trait. The loci of P1M1-27, P3M3-30, and P1M1-20 were identified under all analysis modes of GLM, MLM, GLM + K, and GLM + Q. Conclusion These loci should be compatible to marker assistance selection during A. carmichaelii breeding.
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As a representative variety of hypertoxic herbs, aconite has a very high medicinal value but certain security risks. With the advance of urbanization and industrial transfer and upgrading, traditional geo-authentic regions and emerging regions have gradually coexisted. However, there are significant differences in the geographical distribution, ecological environment and growing cycle in these areas, which may cause intraspecific variations in chemical components, and then affect herb quality and clinical efficacy. Furthermore, there was still lack of in-depth study on characteristic constituents of aconite genuineness. In this paper, an UPLC-Q-TOF-MS was used to investigate the chemical ingredients in Jiangyou aconite, Hanzhong aconite, Butuo aconite and Weishan aconite. The results suggested that using unsupervised PCA method, Jiangyou aconite was similar with Hanzhong aconite in chemical ingredients, but significantly different from ones in emerging regions. Using PLS-DA method, seven characteristic constituents were identified, in which Butuo aconite and Weishan aconite had higher contents of mesaconitine and fuziline, while Jiangyou aconite and Hanzhong aconite had higher contents of hypaconitine, neoline, carmichaeline, and songorine. Based on basic toxicity of each ingredient, we can find that hypaconitine and mesaconitine were the key toxicity factors to distinguish Jiangyou aconite, Butuo aconite, and Weishan aconite. The toxicity of geo-authentic Jiangyou aconite was lower than that of Butuo aconite and Weishan aconite. The lower toxicity in geo-authentic Jiangyou aconite was beneficial for ensuring clinical drug safety, which may be one of the important reasons for the formation of genuine aconite. These results provide new reference data to reveal the formation mechanism of genuine aconite from the perspective of plant secondary metabolite diversity.
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AIM@#The aim of this work was to establish a specific and sensitive method to comprehensively investigate and compare chemical constituents of Fuzi-Gancao herb pair (FG), consisting of Aconitum carmichaelii Debeaux (Fuzi, Chinese) and Roast Radix Glycyrrhizae (Glycyrrhiza glabra L., Gancao, in Chinese) and Fuzi alone to explore the underlying interaction mechanism of FG.@*METHOD@#An ultra-fast liquid chromatography-ion trap/time-of-flight mass spectrometry (UFLC/MS-IT-TOF) method using diazepam as internal standard was developed for the identification and semi-quantitative analysis of the phytochemical constituents of Fuzi and FG. Chromatographic separation was achieved on a UFLC column using a gradient program with 40 mmol·L(-1) ammonium acetate and acetonitrile as the mobile phase.@*RESULTS@#Fifty-one of the sixty compounds, including forty-five C19-diterpenoid alkaloids and six C20-diterpenoid alkaloids were tentatively identified in the extracts of Fuzi and FG through accurate mass measurements and fragmentation patterns. Comparing the contents of these alkaloids in these two extracts, it was found that the diester-diterpenoid alkaloids (DDAs) and the alkylolamine-diterpenoid alkaloids (ADAs) were increased, while the monoester-diterpenoid alkaloids (MDAs) were decreased in the extracts of FG.@*CONCLUSION@#This work provided comprehensive information for the quality control of Fuzi preparations, and the further investigation on the compatibility mechanisms of FG.
Subject(s)
Aconitum , Chemistry , Chromatography, High Pressure Liquid , Methods , Drug Interactions , Drugs, Chinese Herbal , Chemistry , Glycyrrhiza , Chemistry , Mass Spectrometry , Methods , Molecular StructureABSTRACT
Objective: To study the effect of micronutrient fertilizer ratio on the growth and yield of aconite daughter root (the processed daughter root of Aconitum carmichaelii). Methods: Taking A. carmichaelii (Ranunculaceae) as test material, to study the growth and yield of aconite daughter root under the various ratio of Zn, B, Fe, and Mn by the uniform design. Results: To promote the growth and improve the yield of aconite daughter root, the optimum ratio of fertilizer Zn-B-Fe-Mn was 4:1:1:2. Under this ratio, the size of the tuber, the number of the fibrous roots, the length of fibrous roots, the volume of the sub-root, and the dry weight of the tuber, the fibrous roots, and the mother root were significantly or very significantly higher than those under the other fertilizer ratio, while the plant height and the dry weight of leaf were lower than those in the control treatment, with the weight of fibrous roots and the mother roots increased by 24.5% and 25.9% compared with those in the control treatment. The results also showed that there existed negative correlation between the plant height and the yield of per plant, while the dry weight of the fibrous roots and the mother roots showed significantly positive correlation with the yield of per plant. Conclusion: The above mentioned ratio of Zn-B-Fe-Mn as 4:1:1:2 is the best, since it could promote the growth and improve the yield of aconite daughter root more than the other treatment.