Profiling of chemical constituents of Matricarla chamomilla L. by UHPLC-Q-Orbitrap-HRMS and in vivo evaluation its anti-asthmatic activity.

Matricarla chamomilla L. is native to European countries and widely cultivated in China, especially in Xinjiang. It has been used in Uygur medicine for the treatment of cough caused by asthma. In this study, UHPLC-Q-Orbitrap-MS was used to detect and identify the components from the active fraction of M. Chamomile, 64 compounds were identified by combining the standards, related literatures and mass spectrometry fragments, including 10 caffeoyl quinic acids, 38 flavonoids, 8 coumarins, 5 alkaloids and 3 other compounds. Furtherly, the anti-asthma activity of active fraction of M. Chamomile was investigated in OVA-induced allergic asthma rat model. The results showed that the number of EOS in Penh and bronchoalveolar lavage fluid (BALF) in the group of the active fraction of M. Chamomile was significantly lower than that in the model group. Besides, the active fraction of M. Chamomile can significantly reduce the IgE level and increased glutathione peroxidase (GSH-Px) in the serum of OVA-induced rats, and ameliorated OVA-induced lung injury. Hence, M. Chamomile could be used to treat asthma through their in vivo antioxidant and anti-inflammatory effects. This study explored the potential material basis of M. Chamomile for the treatment of asthma.

Anti-inflammatory and anti-rheumatic activities in vitro of alkaloids separated from Aconitum soongoricum Stapf.

The aim of the present study was to investigate the cell proliferation-inhibiting and anti-rheumatic activities of chemical components from Aconitum soongoricum Stapf. Chemical constituents of Aconitum soongoricum Stapf. were separated and purified by silica gel and Sephadex LH-20 chromatography. Structure was identified by spectroscopic technique, and physical/chemical properties were analyzed. The following four compounds were identified: i) Aconitine, ii) songorine, iii) 16, 17-dihydro-12ß, 16ß-epoxynapelline, and iv) 12-epi-napelline. Cell Counting kit-8 assay was performed to assess cell proliferation. ELISA was conducted to determine the cytokine contents, and reverse transcription-quantitative polymerase chain reaction and Western blot analysis were performed to detect the mRNA and protein expression levels. Compared with the lipopolysaccharide (LPS) group, the contents of IL-6, IL-1ß, TNF-α and PGE-2 in the culture supernatant were significantly declined in the leflunomide + LPS and intervention+LPS groups, as well as the mRNA expression levels of HIF-1α, VEGFA and TLR4. Treatments with songorine, benzoylaconine and aconitine (at different concentrations) significantly inhibited the proliferation of HFLS-RA cells. Compared with the LPS group, the contents of PGE-2, IL-6, IL-1ß and TNF-α in the culture supernatant were significantly decreased in the intervention groups, and the mRNA expression levels of TLR4, HIF-1α and VEGFA in the cells in the intervention groups. Songorine, benzoylaconine and aconitine from Aconitum soongoricum Stapf. have anti-rheumatic activities in vitro, which may inhibit the proliferation of HFLS-RA cells, and the underlying mechanisms may be associated with inhibiting the inflammatory cytokine production and downregulating the expression levels of HIF-1α, VEGF and TLR4.

Quantitative proteomics analysis of the treatment of asthma rats with total flavonoid extract from chamomile.

OBJECTIVE: Asthma is a chronic immune disease that has become a serious public health problem. The currently available medications are not ideal because of their limitations and side effects; hence, new target proteins and signaling cascades for precise and safe therapy treatment are needed. This work established an ovalbumin-induced asthma rat model and treated it with total flavonoid extract from the Xinjiang chamomile. The proteins that were differentially expressed in the chamomile extract-treated asthmatic rats and the asthma and healthy rat groups were identified using isobaric tagging followed by LC-MS/MS. Kyoto encyclopedia of genes and genomes pathway analysis of the differentially expressed proteins was performed. RESULTS: Pathways involved in purine metabolism, herpes simplex infection, and JNK phosphorylation and activation mediated by activated human TAK1 were enriched, indicating the intrinsic links between the mechanism of asthma development and treatment effects. Furthermore, we constructed a protein-protein interaction network and identified KIF3A as a potential target protein of chamomile extract that affected the Hedgehog signaling pathway. CONCLUSIONS: This study may provide new insights into the pathogenesis of asthma and reveal several proteins and pathways that could be exploited to develop novel treatment approaches.

Anti-rheumatic effects of Aconitum leucostomum Worosch. on human fibroblast-like synoviocyte rheumatoid arthritis cells.

The aim of the present study was to investigate the effects of Aconitum leucostomum Worosch. crude drug, processed products and monomer components on human fibroblast-like synoviocyte rheumatoid arthritis (HFLS-RA) cells, and its associated mechanisms. Following drug treatment, cell proliferation was assessed using a Cell Counting Kit-8 assay. Cellular apoptosis and cell cycle were evaluated using flow cytometry. Levels of hypoxia-inducible factor 1α (HIF-1α), vascular endothelial growth factor (VEGF) and toll-like receptor 4 (TLR4) mRNA and protein were evaluated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis, respectively. Levels of pro-inflammatory cytokines were evaluated using ELISA. Analysis of cell proliferation indicated that crude drug and processed products markedly inhibited the cell proliferation. Compared with the control group, the apoptosis rates were significantly elevated in all treatment groups (all P<0.05). Furthermore, the proportion of cells in G0/G1 phase was significantly decreased in all treatment groups compared with the control group (all P<0.05). RT-qPCR and western blotting indicated that, compared with the control group, mRNA and protein expression levels of HIF-1α, and TLR4 were significantly downregulated in all treatment groups (P<0.05). The mRNA and protein expression levels of VEGF in all treatment groups were decreased compared with those in the control group, but the difference was not significant. Results from ELISA demonstrated that the levels of interleukin (IL)-6, IL-1ß and tumor necrosis factor-α in the cell culture supernatant were all significantly decreased following drug treatment in HFLS-RA cells (all P<0.05). Therefore, A. leucostomum Worosch. crude drug, processed products and monomer components may exert anti-rheumatic effects on HFLS-RA cells, inhibiting cell proliferation and enhancing cellular apoptosis. These effects may be attributable to the downregulated expression of HIF-1α and TLR4, as well as decreased levels of pro-inflammatory cytokines.

Assessment of in vitro cardiotoxicity of extract fractions and diterpene alkaloids from Aconitum leucostomum Worosch: A short communication.

Aconitum leucostomum Worosch is a traditional Chinese medicine (TCM) and has a broad spectrum of health effects, but with a narrow therapeutic window. It is important to identify both the therapeutic ingredients and the toxic components to better utilize this TCM. The present study investigated the cardiotoxicity of the selected compounds in Aconitum leucostomum Worosch. The effects of extract of A. leucostomum Worosch and the isolated compounds on cardiocardiomyocytes were evaluated in vitro. Five known compounds in this TCM, including three C18-diterpene alkaloids, lappaconitine (2), N-deacetyllappaconitine (3), and ranaconitine (5), and two C19-diterpene alkaloids, delvestidine (1) and anthranoyllycoctonine (4), were isolated from A. leucostomum Worosch. The cardiotoxicity of these components and extract fractions, as measured by lactate dehydrogenase release and apoptosis, was ranked as follows, in descending order: delvestidine>anthranoyllycoctonine>pH 4 fraction>pH 8 fraction>aconitine>N-deacetyllappaconitine>ranaconitine>lappaconitine. The cytotoxicity of these compounds was shown to be dose-dependent, with delvestidine (1) and anthranoyllycoctonine (4) being the two most toxic compounds to cardiomyocytes in our assays. These results provide a basis for future rational use of this TCM, reducing side effects while retaining therapeutic effects.

Assessment of genetic characteristics of Aconitum germplasms in Xinjiang Province (China) by RAPD and ISSR markers.

Aconitum is a medicinal treasure trove that grows extensively on fertile pastures in Xinjiang Province (China); however, its molecular genetic characteristics are still poorly studied. We studied Aconitum kusnezoffii Reichb., Aconitum soongaricum Stapf., Aconitum carmichaelii Debx. and Aconitum leucostomum Worosch, using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) techniques, to evaluate their genetic relationship and potential medicinal value. Our results showed that A.kusnezoffii Reichb. and A.soongaricum Stapf. have close genetic relationship and cluster together. Polymorphism rates of 97.25% and 98.92% were achieved by using 15 RAPD and 15 ISSR primers, respectively. Based on Nei's gene diversity (H) and Shannon's index (I), the inter-population diversity (Hs ) was higher when compared with the intra-population diversity (Hp ). Among the three Aconitum populations, the coefficient of gene differentiation (Gst ) was 0.4358 when evaluated by RAPD and 0.5005 by ISSR. The genetic differentiation among the three Aconitum populations was highly significant, suggesting low gene flow (Nm ). This was confirmed by the estimates of gene flow (Nm = 0.6473 and Nm = 0.4991, based on ISSR and RAPD data, respectively). Comparing the RAPD and ISSR results, the two DNA markers proved similarly effective in the assessment of the genetic characteristics of the studied Aconitum populations and could be used for reliable fingerprinting and mapping in studies on Aconitum diversity in view of Aconitum suitability for development and protection.

[Research on the effect of Aconitum soongaricum and its processed products on AA rat].

OBJECTIVE: To observe the effect of Aconitum soongaricum and its processed products on adjuvant arthritis (AA) rat and its mechanism. METHODS: Rats were radomly divided into normal group, model group, positive group, Aconitum soongaricum and its processed products (including Pharmacopoeia method, document method, Kazak method three processed products), which divided into high dose, middle dose, low dose groups (30.65, 15.32, 7.66 mg/kg); Except normal group, prepared AA rat model by using Freund's complete Adjuvant (FCA) for the rest groups; Each group was administered for 23 days after the rat model was established, observed rats ankle swelling degree and arthritis index. Determined content of interleukin-1beta (IL-1beta) and cortisol (Cort) in serum by ELISA method. RESULTS: (1) The swelling degree of model group was higher than that of normal group (P < 0.01). Compared with model group, the swelling degree and AI of Aconitum soongaricum and all of its processed products groups were lower (P < 0.01 or P < 0.05). (2) The content of IL-1beta and Cort in serum of model group was higher than that of normal group (P < 0.01). Compared with model group, the content of Cort in serum of positive group, Aconitum soongaricum groups (high dose, middle dose, low dose), its Pharmacopoeia method processed product groups (high dose, middle dose) and its document method processed product groups (high dose) were lower (P < 0.01); Compared with model group, the content of IL-1beta in serum of positive group, Aconitum soongaricum groups (high dose, middle dose, low dose), its Pharmacopoeia method processed product groups (high dose, middle dose, low dose) its document method processed product group (high dose) and its Kazak method processed product group (middle dose) were lower (P < 0.01 or P < 0.05). CONCLUSION: Aconitum soongaricum and all of its processed products groups are proved with anti-inflammatory effect, which possible mechanism would be restraining inflammation cell factor such as IL-1beta and Cort in serum of AA rat.

[Studies of preparing procedures of Yuhuangliang].

OBJECTIVE: To optimize preparing procedures of Yuhuangliang. METHODS: The single factor experimets and the L9 (3(4)) orthogonal design were used to optimize the preparing procedures of Yuhuangliang with contents of berberine and total alkaloids as evalustion index. RESULTS: The optimal preparing method was Rhizoma Coptidis adding amount of 10 mL (0.2 g/mL) juice of wuzhuyu mixed thoroughly, baking at 100 degrees C, for 15 min. CONCLUSION: The preparing procedure is effective, stable and reliable.