Anti-inflammatory diterpenoid alkaloids from Aconitum tanguticum (Maxim.) Stapf.

Fifteen undescribed diterpenoid alkaloids and seven known analogs were coisolated from the whole plant Aconitum tanguticum (Maxim.) Stapf. Their structures were elucidated based on spectroscopic methods. Among them, tangirine A was a heteratisine-hetidine-type bis-diterpenoid alkaloid, tanguticinines A-F were six hetidine-hetisine-type bis-diterpenoid alkaloids, tanguticinine G was one hetidine-atisine-type bis-diterpenoid alkaloid, and N-oxide anthoroidine B, 5-deoxyanthoridine B and N-oxide 5-deoxyanthoroidine B were three unusual hetidine-rearranged hetisine-type bis-diterpenoid alkaloids. In the bioassay, thirteen compounds showed some inhibitory effects on the secretion of NO and TNF-α in LPS-treated RAW 264.7 cells, with IC50 values of 67.56 µM-683.436 µM.

The anti-inflammatory mechanism of Aconitum tanguticum based on network pharmacology and molecular docking / 中国药理学通报

Aim To study the potential molecular anti-inflammatory mechanism of Aconitum tanguticum based on network pharmacology methods,molecular docking technology and cell experiment. Methods The active ingredients targets and disease targets of Aconitum tanguticum were collected through literature and database. The common targets were utilized by mixture of them and the core targets were obtained by constructing the protein protein interaction(PPI)network. Then the component-target-disease network diagram was constructed. The gene ontology(GO)analysis and Kyoto encyclopedia of genes and genomes(KEGG)analysis were performed for common targets. AutoDock Vina(1.1.2)software was utilized for combining some of the core targets and the diterpenoid alkaloids in the chemical components of Aconitum tanguticum. Finally,the influence of alcoholic extract of Aconitum tanguticum(ATS)on RAW264.7 cell inflammation model was preliminarily verified by MTT assay,Griess reagent and realtime RT-PCR. Results A total 17 main active ingredients were obtained from literature and 284 common targets were obtained via intersecting with disease targets. Altogether 108 pathways were screened by KEGG enrichment,mainly including PI3K-Akt,Ras,MAPK and HIF-1. Molecular docking results indicated that the active ingredients of Aconitum tanguticum had a high affinity with the core target to be docked. In vitro experiment suggested that ATS treatment inhibited LPS-induced NO production and iNOS mRNA in RAW264.7 cells. Realtime RT-PCR detection suggested that ATS played an anti-inflammatory effect by regulating the PI3K-Akt signaling pathway. Conclusions Aconitum tanguticum exerts anti-inflammatory effects through PI3K-Akt pathways,which provides the scientific basis for better promoting the development of Aconitum tanguticum.

[HPLC fingerprint of Tibetan medicinal plant ″Bangga″ and chemical components analysis of its two original plants by LC-MS/MS].

To obtain the chemical profile of Tibetan medicinal plant ″Bangga″, the present study established the HPLC fingerprint of ″Bangga″ and inferred common chemical constituents of its two original plants, Aconitum tanguticum and A. naviculare by LC-MS. The HPLC analysis was performed on a Kromasil 100 C_8 column(4.6 mm×250 mm, 5 µm) with acetonitrile(A)-0.1% formic acid in water(B) as mobile phase in a gradient elution mode. Besides, the flow rate was set at 1 mL·min~(-1) and the column temperature was 35 ℃. The detection wavelength was set at 255 nm and the injection volume was 10 µL. Seventeen batches of ″Bangga″ samples were analyzed and the HPLC fingerprint was established under the above conditions. Similarity evaluation was performed using Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012). As a result, 16 common peaks were selec-ted and the similarity values of 17 batches of ″Bangga″ were in the range of 0.702-0.966. Furthermore, one batch of A. tanguticum and one batch of A. naviculare were analyzed by LC-MS/MS and 74 common compounds were inferred, including 10 phenolic acids, 26 flavonoids, and 38 alkaloids. The established method, with good separation and strong specificity, is simple and feasible, and can be used for the quality control of ″Bangga″ and identification of its two original plants. A. tanguticum and A. naviculare are similar in chemical composition and component content, but are quite different in the content of flavonoids.

Tangutidines A-C, Three Amphoteric Diterpene Alkaloids from Aconitum tanguticum.

Three new diterpene alkaloids, tangutidines A-C (1-3), and four known alkaloids (4-7) were isolated from the whole plant of Aconitum tanguticum, from which amphoteric diterpene alkaloids (1-3) were obtained for the first time. The structures of 1-3 were elucidated by detailed interpretation of spectroscopic data, including MS and NMR data. All of them were evaluated for their cytotoxic activities.

Anti-acetylcholinesterase Active Ingredients from Aconitum tanguticum by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To identify the anti-acetylcholinesterase active ingredients in <italic>Aconitum tanguticum</italic>， so as to lay the foundation for finding new anti-Alzheimer's disease （AD） drugs. Method:The anti-acetylcholinesterase active fractions of <italic>A. tanguticum</italic> were screened by the modified Ellman's method， and the chemical composition of the active fraction was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. The chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column （2.1 mm×50 mm， 1.7 μm） with acetonitrile （A）-0.4% ammonia aqueous solution （B） as mobile phase for gradient elution， and the column temperature was set at 30 ℃ with the flow rate of 0.4 mL·min^-1. Phase A of the dichloromethane fraction changed with time as follows：0-3 min， 5%A； 3-7 min， 5%-20%A； 7-11.5 min， 20%-33%A； 11.5-15.5 min， 33%-50%A； 15.5-20.5 min， 50%-80%A； 20.5-23 min， 80%-85%A； 23-25 min， 85%-95%A. Phase A of the <italic>n</italic>-butanol fraction changed with time as follows：0-2 min， 5%A； 2-8 min， 5%-20%A； 8-11 min， 20%-33%A； 11-15 min， 33%-95%A. Mass spectrometry was performed on electrospray ionization， data were collected in positive ion mode， and the detection range was <italic>m</italic>/<italic>z</italic> 100-1 500. Result:Both the dichloromethane and <italic>n</italic>-butanol fractions had a certain inhibitory effect on acetylcholinesterase， their half inhibitory concentration （IC₅₀） values were （64±4.4） mg·L^-1 and （85.7±3.8） mg·L^-1， respectively. By UPLC-Q-TOF-MS/MS analysis， a total of 21 alkaloids were identified from the dichloromethane fraction， and 11 alkaloids were identified from <italic>n</italic>-butanol fraction. Guan-fu base Ⅰ， found in both fractions， was first discovered in <italic>A. tanguticum</italic>. Conclusion:Diterpene alkaloids are the main anti-acetylcholinesterase substances of <italic>A. tanguticum</italic>， which is worth further exploration.

A novel dimeric diterpenoid alkaloid from the Tibetan medicinal plant Aconitum tanguticum (Maxim.) Stapf / 药学学报

Two dimeric diterpenoid alkaloids were isolated from the whole plant of Aconitum tanguticum (Maxim.) Stapf and their structures were elucidated by extensive analysis of 1D, 2D-NMR and HR-MS data. One is a new compound and named tanguticurine A (1), and the other is the known compound anthoroidine B (2); both were isolated from this plant for the first time. The antiviral activity of compounds 1 and 2 against HCV and EV71 were also evaluated. It was found that compound 1 had a good inhibitory effect on HCV and EV71 with EC50 values of 15.5 and 9.7 μmol·L-1, respectively, and showed low cytotoxicity. Therefore, compound 1 is a good antiviral lead compound and deserves further study.

HPLC fingerprint of Tibetan medicinal plant ″Bangga″ and chemical components analysis of its two original plants by LC-MS/MS / 中国中药杂志

To obtain the chemical profile of Tibetan medicinal plant ″Bangga″, the present study established the HPLC fingerprint of ″Bangga″ and inferred common chemical constituents of its two original plants, Aconitum tanguticum and A. naviculare by LC-MS. The HPLC analysis was performed on a Kromasil 100 C_8 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.1% formic acid in water(B) as mobile phase in a gradient elution mode. Besides, the flow rate was set at 1 mL·min~(-1) and the column temperature was 35 ℃. The detection wavelength was set at 255 nm and the injection volume was 10 μL. Seventeen batches of ″Bangga″ samples were analyzed and the HPLC fingerprint was established under the above conditions. Similarity evaluation was performed using Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012). As a result, 16 common peaks were selec-ted and the similarity values of 17 batches of ″Bangga″ were in the range of 0.702-0.966. Furthermore, one batch of A. tanguticum and one batch of A. naviculare were analyzed by LC-MS/MS and 74 common compounds were inferred, including 10 phenolic acids, 26 flavonoids, and 38 alkaloids. The established method, with good separation and strong specificity, is simple and feasible, and can be used for the quality control of ″Bangga″ and identification of its two original plants. A. tanguticum and A. naviculare are similar in chemical composition and component content, but are quite different in the content of flavonoids.

Characterization of the complete chloroplast genome of the Tangut monkshood Aconitum tanguticum (Ranunculales: Ranunculaceae).

The Tangut monkshood (Aconitum tanguticum) is a perennial herb with high medicinal values. Here, its chloroplast genome was assembled from Illumina sequencing reads. The circular genome is 157,114 bp long with an A + T-biased nucleotide composition, and comprises a pair of inverted repeat (IR) regions (26,255 bp), separated by a large single-copy (LSC) region (87,559 bp) and a small single-copy (SSC) region (17,045 bp). It encodes a total of 112 gene species, with 19 of them being completely or partially duplicated and 18 of them harboring one or two introns. Phylogenetic analysis recovered two major clades of the genus Aconitum.

Diterpenoid alkaloids from the whole plant of Aconitum tanguticum (Maxim.) Stapf.

Five undescribed diterpenoid alkaloids tanguticulines A-E and two undescribed amide compounds 5-methoxy-N-Salicylanthranilic acid methyl ester and 3, 5-dimethoxy-4- hydroxycinnamamide-4-O-ß-D-glucopyranoside, were isolated from the whole plant of Aconitum tanguticum (Maxim.) Stapf. Their structures were assigned by detailed analysis of MS and NMR spectroscopic data. Compounds 1 and 5 were evaluated for their antivirus activities against influenza a (H1N1) virus in vitro. Both of them showed obvious inhibitory effect on the cytopathic changes induced by H1N1, with IC50 values of 2.9 µg mL-1 and 2.4 µg mL-1, respectively.

The total alkaloids of Aconitum tanguticum protect against lipopolysaccharide-induced acute lung injury in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Aconitum tanguticum has been widely used as a remedy for infectious diseases in traditional Tibetan medicine in China. The total alkaloids of Aconitum tanguticum (TAA) are the main active components of Aconitum tanguticum and have been demonstrated to be effective in suppressing inflammation. Our aim was to investigate the protective effects of TAA on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats. MATERIALS AND METHODS: TAA was extracted in 95% ethanol and purified in chloroform. After vacuum drying, the TAA powder was dissolved in dimethyl sulfoxide. Adult male Sprague-Dawley rats were randomly divided into six groups. Rats were given dexamethasone (DXM, 4 mg/kg) or TAA (60 mg/kg, 30 mg/kg) before LPS injection. The PaO2and PaO2/FiO2 values, lung wet/dry (W/D) weight ratio and histological changes in lung tissue were measured. The cell counts, protein concentration, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) in bronchoalveolar lavage fluid (BALF), and myeloperoxidase (MPO) activity in lung tissue were determined at 6, 12 or 24 h after LPS treatment. In addition, the NF-κ B activation in lung tissue was analyzed by western blot. RESULTS: In ALI rats, TAA significantly reduced the lung W/D ratio and increased the value of PaO2 or PaO2/FiO2 at 6, 12 or 24 h after LPS challenge. TAA also reduced the total protein concentration and the number of total cells, neutrophils or lymphocytes in BALF. In addition, TAA decreased MPO activity in the lung and attenuated histological changes in the lung. Furthermore, TAA inhibited the concentration of TNF-α, IL-6 and IL-1ß in BALF at 6, 12 or 24 h after LPS treatment. Further study demonstrated that TAA significantly inhibited NF-κ B activation in lung tissue. CONCLUSIONS: The current study proved that TAA exhibited a potent protective effect on LPS-induced ALI in rats through its anti-inflammatory activity.

Two new compounds from Aconitum tanguticum.

A phytochemical investigation on the whole plant of Aconitum tanguticum (Ranunculaceae) resulted in the isolation and characterization of two new phenylpropanoid glycosides (1 and 2). Their structures were elucidated as 4-hydroxyphenethoxy-8-O-ß-d-[6-O-(4-O-ß-d-glucopyranosyl)-sinapoyl]-glucopyranoside (1) and 3,4-dimethoxy-trans-cinnamic acid-9-O-ß-d-allopyranoside (2) on the basis of spectroscopic data (1D NMR, 2D NMR, and MS) and comparison with the literature data.