Comparison of Anti-inflammatory Activities of Six Curcuma Rhizomes: A Possible Curcuminoid-independent Pathway Mediated by Curcuma phaeocaulis Extract

We aimed to compare the anti-inflammatory activities of six species of Curcuma drugs using adjuvant arthritis model mice. When orally administered 1 day before the injection of adjuvant, the methanol extract of Curcuma phaeocaulis significantly inhibited paw swelling and the serum haptoglobin concentration in adjuvant arthritis mice. Also when orally administered 1 day after the injection of adjuvant, the methanol extract of Curcuma phaeocaulis significantly inhibited paw swelling. Other Curcuma species (Curcuma longa, Curcuma wenyujin, Curcuma kwangsiensis, Curcuma zedoaria and Curcuma aromatica) had no significant inhibitory effects on adjuvant-induced paw swelling. Cyclooxygenase (COX)-2 activity was significantly inhibited by the methanol extract of C. phaeocaulis. Curcuminoids' (curcumin, bis-demethoxycurcumin and demethoxycurcumin) were rich in C. longa, but less in C. phaeocaulis and C. aromatica, not in C. wenyujin, C. kwangsiensis and C. zedoaria, suggesting that curcuminoids' contents do not relate to inhibition of arthritis swelling. Therefore, C. phaeocaulis may be a useful drug among Curcuma species for acute inflammation, and the active constituents of C. phaeocaulis are not curcuminoids

Curdione Plays an Important Role in the Inhibitory Effect of Curcuma aromatica on CYP3A4 in Caco-2 Cells

Curcuma aromatica is a plant belonging to genus Curcuma of family Zingiberaceae and is widely used as supplements in Japan. Rhizomes of C. aromatica have curcumin as a major yellow pigment and curdione as a main ingredient of essential oils. In this study, we investigated the affect of C. aromatica on CYP3A4 using 1α,25-(OH)2-D3-treated Caco-2 clone cells. Caco-2 cells were treated with methanol extract (0.1 mg ml−1), its hexane soluble fraction (0.1 mg ml−1), curcumin (4 μM) and curdione (20 μM) for 72 hours. Nifedipine was used as a substrate of CYP3A4. Methanol extract, hexane fraction and curdione inhibited the formation of oxidized nifedipine by 50–70%, and curcumin showed no effect. The IC50s of methanol extract, hexane fraction and curdione to oxidized nifedipine formation were 21, 14 and 3.9 μg ml−1 (16.9 μM), respectively. The content of curdione in methanol extract was 11.4%. Moreover, all of methanol extract, hexane fraction and curdione decreased CYP3A4 protein expression but had no affect on CYP3A4 mRNA expression. Our results showed that these drugs further decreased the CYP3A4 protein expression level after the protein synthesis was inhibited by cychroheximide. These findings suggest that curdione plays an important role in the CYP3A4 inhibitory activity of C. aromatica and curdione might inhibit the activity by accelerating the degradation of CYP3A4

Mass spectrometry imaging of the capsaicin localization in the capsicum fruits

We succeeded in performing mass spectrometry imaging (MSI) of the localization of capsaicin in cross-sections of the capsicum fruits at a resolution of 250 µm using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Post source decay of protonated capsaicin ion revealed structural information of the corresponding acid amide of vanillylamide and C9 chain fatty acid. MALDI-TOF-MSI confirmed that localization of capsaicin in the placenta is higher than that in the pericarp. In addition, it revealed no localization of capsaicin in seed and the higher localization of capsaicin at placenta surface compared with that in the internal region. A quantitative difference was detected between localizations of capsaicin at placenta, pericarp and seed in the capsicum fruits. This imaging approach is a promising technique for rapid quality evaluation general food as well as health food and identification of medicinal capsaicin in plant tissues

Anti-Trypanosoma cruzi activity of Coptis rhizome extract and its constituents

Background:Current therapeutic agents, including nifurtimox and benznidazole, are not sufficiently effective in the chronic phase of Trypanosoma cruzi infection and are accompanied by various side effects. In this study, 120 kinds of extracts from medicinal herbs used for Kampo formulations and 94 kinds of compounds isolated from medicinal herbs for Kampo formulations were screened for anti-T. cruzi activity in vitro and in vivo.Methods:As an experimental method, a recombinant protozoan cloned strain expressing luciferase, namely Luc2-Tulahuen, was used in the experiments. The in vitro anti-T. cruzi activity on epimastigote, trypomastigote, and amastigote forms was assessed by measuring luminescence intensity after treatment with the Kampo extracts or compounds. In addition, the cytotoxicity of compounds was tested using mouse and human feeder cell lines. The in vivo anti-T. cruzi activity was measured by a murine acute infection model using intraperitoneal injection of trypomastigotes followed by live bioluminescence imaging.Results:As a result, three protoberberine-type alkaloids, namely coptisine chloride, dehydrocorydaline nitrate, and palmatine chloride, showed strong anti-T. cruzi activities with low cytotoxicity. The IC50 values of these compounds differed depending on the side chain, and the most effective compound, coptisine chloride, showed a significant effect in the acute infection model.Conclusions:For these reasons, coptisine chloride is a hit compound that can be a potential candidate for anti-Chagas disease drugs. In addition, it was expected that there would be room for further improvement by modifying the side chains of the basic skeleton

Scientific basis for the anti-dementia drugs of constituents from Ashwagandha (Withania somnifera)(Chemical & Pharmacological study)

Although Ashwagandha (root of Withania somnifera) has been used for multi purposes but mainly as a tonic in the traditional Indian Ayurvedic medicine, this review focused on nootropic effects of Ashwagandha itself and isolated compounds from it. Several reports showed sitoindosides VII-X as active compounds for cognitive enhancer. However, our study revealed other active constituents, withanolide A, withanoside IV and withanoside VI, could improve Aβ(25-35) induced memory impairment, neuronal atrophy and synaptic loss in the cerebral cortex and the hippocampus. Treatment with Aβ(25-35) induced axonal and dendritic atrophy, and pre-synaptic and post-synaptic losses also in cultured rat cortical neurons. Subsequent treatment with withanolide A, withanoside IV and withanoside VI induced significant reconstruction of pre-synapses and post-synapses, in addition to regeneration of both axons and dendrites in the neurons. Withanolide A, withanoside IV and withanoside VI are therefore important candidates for the therapeutic treatment of neurodegenerative diseases, as it is able to reconstruct neuronal networks.Century COE program, Toyama Medical and Pharmaceutical Universit

Systematic pharmacognostical study on Panax drugs and Curcuma drugs : Phylogenetic analysis, molecular authentication and quality evaluation

We proposed pharmacognostical studies in the prime of molecular biology, citing the systematic studies of Panax drugs and Curcuma drugs. Each study was composed of three approaches, phylogenetic analysis of plants based on nuclear 18S rRNA and chloroplast trnK gene sequences, molecular authentication of herbal drugs, and quality evaluation on bioactive chemical constituents or pharmacological effect. Parsimony analysis of the combined trnK-18S rRNA gene sequence data yielded a well-resolved phylogeny within genus Panax. Based on species-specific sequences of the 2 genes, all the Panax drugs could be identified, furthermore, multiplex amplification refractory mutation system assay was developed for the authentication of 5 important drugs. Quantitative analysis on 11 saponins revealed that each taxon possessed its own characteristic pattern. The trnK/18S rRNA gene sequences could be used not only for an ultimate authentication but also for a speculation of the chemical constituent pattern that affects pharmacological effects. By the same molecular analysis as genus Panax, the potential method for identification of Chinese and Japanese Curcuma species was developed, making it possible to identify Curcuma drugs unambiguously.Panax属植物及びCurcuma属植物に由来する人参類生薬及び鬱金類生薬に関する研究を例にして, 分子生物学全盛期の現代における生薬学的研究を提唱した。研究は, 核18SrRNA遺伝子及び葉緑体trnK遺伝子の塩基配列に基づく植物の分子系統学的解析, 遺伝子多型に基づく生薬の同定, 薬理活性成分または薬理活性に基づく生薬の品質評価から構成される。2遺伝子の塩基配列に基づいて構築した系統樹からPanax属植物の系統関係が明確に整理され, 一方各分類群に固有な塩基配列からすべての同属由来の生薬が同定された。重要な5種類の生薬を簡便に同定する方法としてMARMS法を開発した。Panax属12分類群に由来する生薬について, 11サポニン成分の定量分析を行った結果, 分類群固有の含有パターンが見出された。したがって, 遺伝子の塩基配列を決定すれば, 生薬の同定のみならず, 各種活性を有する成分の組成をも推定できることが示唆された。同様に中国及び日本産Curcuma属植物の遺伝子解析を行い, 鬱金類生薬の正確な同定を可能にした。5種類の生薬の駆於血作用を比較する目的で, 血管リング標本を用いて血管作動性を検討した結果, 全生薬のメタノールエキスに強いNO非依存性の血管弛緩作用, 熱水エキスにそれより弱い弛緩作用が認められた。これは, 熱水エキスにはメタノール可溶性画分に起因する血管弛緩作用の他, 多糖類に起因する血管収縮作用が認められることによるものである。この弛緩作用は基源種により異なり, また日本産ガジュツのみ他種と同様のNO非依存性の血管弛緩作用に加えて, NO依存性の血管弛緩作用が認められた。ここで概説した一連の研究は, 生薬の標準化とそれらの効率的な利用の点から重要になるものと考えられる。relation: isVersionOf: http://ci.nii.ac.jp/naid/10014223097/en/Century COE Program, Toyama Medical and Pharmaceutical Universit

Authentication of Rhei Rhizoma(Material study)

Rhei Rhizoma (Dahuang in Chinese) is widely known as a purgative and antiinflammatory agent. In the Japanese Pharmacopoeia, Rhei Rhizoma is prescribed for 4 Rheum species, Rheum palmatum, R. tanguticum, R. officinale, and R. coreanum, while the first 3 species are prescribed for Dahuang in the Chinese Pharmacopoeia. Due to the morphologic similarity of the aerial parts, the taxonomy of this genus and the correct identification of Rheum species and their derivative drugs are very difficult. To resolve taxonomic problems of the genus Rheum and develop an ultimate identification method for plants and drugs, molecular analysis of the chloroplast matK gene and nuclear 18S ribosomal RNA gene were performed on 9 species. The sequence comparison of the matK gene revealed that most species had variable sequences not only inter- but also intraspecies. However, the specimens of the same species belonged to the same subclade in the phylogenetic tree constructed based on matK gene sequences, except for R. palmatum, in which specimens belonged to 3 subclades related to their production areas. The nucleotide differences at positions 587,707, and 838 distinguished official species from others, while specific nucleotides at positions 367 and 937 became identification markers for R. palmatum, R. tanguticum, and R. officinale. Moreover, three groups of R. palmatum, each belonging to 3 subclades, were characterized by the nucleotides at positions 619,769,883, and 1061. On the basis of the above marker nucleotides, a convenient and efficient identification method employing Polymerase Chain Reaction-Restriction Fragment Length Polymorphism and Amplification Refractory Mutation System analyses was further developed. The procedure enabled us to identify the botanic origins of 22 drug samples of Rhei Rhizoma. The matK gene sequence was valuable in identifying Rheum species and Rhei Rhizoma and in predicting their production areas, and could be used as an index for quality evaluation of Rhei Rhizoma