Effects of HIIT associated with Coutoubea spicata supplementation on tissue and oxidative damage biomarkers in Wistar rats / Efectos del HIIT asociado a la suplementación de Coutoubea spicata en los biomarcadores del tejido y daño oxidativo en ratas Wistar

SUMMARY: This study aimed to evaluate the effects of six weeks of HIIT on tissue and oxidative damage markers in rats supplemented with Coutoubea spicata fraction. Thirty-two male Wistar rats were divided into 4 groups: Baseline (GB); supplemented with 100 mg/kg of Coutoubea spicata fraction (GSCS); exercised for 6 weeks with the HIIT protocol (GH); supplemented with 100 mg/kg of Coutoubea spicata fraction + HIIT for 6 weeks (GHCS). Exercised animals performed the HIIT protocol (2 x 2). Tissue damage CK, LDH, ALT and AST markers in plasma were analyzed, as well as oxidative stress MDA and SH biomarkers in plasma and in cardiac, hepatic and muscle tissues. The results showed that CK, LDH, AST and ALT enzymes showed increase in GH when compared to GB (p<0.0001). However, CK, AST and ALT markers reduced their concentrations in GHCS when compared to GH (p<0.0001), indicating that Coutoubea spicata supplementation attenuated the damage in muscle and liver tissues induced by HIIT. Plasma, liver and muscle MDA showed increase in GH after HIIT sessions; however, when compared to GHCS, it showed reduced levels (p<0.0001). SH was elevated in the GH group when compared to GB in plasma and liver tissues (p<0.0001); in contrast, reduction in GHCS when compared to GH was observed in plasma, liver and cardiac tissues, demonstrating the redox effect of HIIT on some tissues. Thus, our findings showed that Coutoubea spicata has antioxidant activity, reducing oxidative damage markers and consequently tissue damage in healthy Wistar rats after HIIT protocol, but it also demonstrated redox balance after analyzing oxidative stress markers.

RESUMEN: Este estudio tuvo como objetivo evaluar los efectos de HIIT en los marcadores de daño tisular y oxidativo en ratas suplementadas con Coutoubea spicata durante seis semanas. Treinta y dos ratas Wistar macho se dividieron en 4 grupos: línea de base (GB); suplementados con 100 mg/kg de fracción de Coutoubea spicata (GSCS); ejercitados durante 6 semanas con el protocolo HIIT (GH); suplementado con 100 mg/kg de fracción de Coutoubea spicata + HIIT durante 6 semanas (GHCS). Los animales ejercitados realizaron el protocolo HIIT (2x2). Se analizaron los marcadores de daño tisular CK, LDH, ALT y AST en plasma, así como los biomarcadores de estrés oxidativo MDA y SH en plasma y en tejidos cardiaco, hepático y muscular. Los resultados indicaron que las enzimas CK, LDH, AST y ALT mostraron aumento en GH en comparación con GB (p<0,0001). Sin embargo, los marcadores CK, AST y ALT redujeron sus concentraciones en GHCS en comparación con GH (p<0,0001), lo que indica que la suplementación con Coutoubea spicata atenuó el daño en los tejidos musculares y hepáticos inducido por HIIT. La MDA de plasma, hígado y músculo mostró un aumento en la GH después de las sesiones de HIIT; sin embargo, en comparación con GHCS, mostró niveles reducidos (p<0,0001). Se observó SH elevado en el grupo de GH en comparación con GB en plasma y tejidos hepáticos (p<0,0001); en contraste, se observó una reducción en GHCS en comparación con GH en plasma, hígado y tejidos cardíacos, lo que demuestra el efecto redox de HIIT en algunos tejidos. Por lo tanto, nuestros hallazgos mostraron que Coutoubea spicata tiene actividad antioxidante, con reducción de los marcadores de daño oxidativo y, en consecuencia, el daño tisular en ratas Wistar sanas después del protocolo HIIT, pero además demostró el equilibrio redox después de analizar los marcadores de estrés oxidativo.

Chemistry, Pharmacology and Therapeutic Potential of Swertiamarin - A Promising Natural Lead for New Drug Discovery and Development.

Swertiamarin, a seco-iridoid glycoside, is mainly found in Enicostemma littorale Blume (E. littorale) and exhibits therapeutic activities for various diseases. The present study aimed to provide a review of swertiamarin in terms of its phytochemistry, physicochemical properties, biosynthesis, pharmacology and therapeutic potential. Relevant literature was collected from several scientific databases, including PubMed, ScienceDirect, Scopus and Google Scholar, between 1990 and the present. This review included the distribution of swertiamarin in medicinal plants and its isolation, characterization, physicochemical properties and possible biosynthetic pathways. A comprehensive summary of the pharmacological activities, therapeutic potential and metabolic pathways of swertiamarin was also included after careful screening and tabulation. Based on the reported evidence, swertiamarin meets all five of Lipinski's rules for drug-like properties. Thereafter, the physicochemical properties of swertiamarin were detailed and analyzed. A simple and rapid method for isolating swertiamarin from E. littorale has been described. The present review proposed that swertiamarin may be biosynthesized by the mevalonate or nonmevalonate pathways, followed by the seco-iridoid pathway. It has also been found that swertiamarin is a potent compound with diverse pharmacological activities, including hepatoprotective, analgesic, anti-inflammatory, antiarthritis, antidiabetic, antioxidant, neuroprotective and gastroprotective activities. The anticancer activity of swertiamarin against different cancer cell lines has been recently reported. The underlying mechanisms of all these pharmacological effects are diverse and seem to involve the regulation of different molecular targets, including growth factors, inflammatory cytokines, protein kinases, apoptosis-related proteins, receptors and enzymes. Swertiamarin also modulates the activity of several transcription factors, and their signaling pathways in various pathological conditions are also discussed. Moreover, we have highlighted the toxicity profile, pharmacokinetics and possible structural modifications of swertiamarin. The pharmacological activities and therapeutic potential of swertiamarin have been extensively investigated. However, more advanced studies are required including clinical trials and studies on the bioavailability, permeability and administration of safe doses to offer swertiamarin as a novel candidate for future drug development.

Molecular docking, antiproliferative and anticonvulsant activities of swertiamarin isolated from Enicostemma axillare.

Present study aimed for molecular docking, antiproliferative and anticonvulsant activities of swertiamarin isolated from the successive methanol extract of Enicostemma axillare. Molecular docking of swertiamarin on telomerase targets (PDB ID: 5UGW, 3DU6 and 4ERD), followed by antiproliferative activity on HEp2 and HT-29 cells by MTT and SRB assays. Also tested for anticonvulsant activity by pentylenetetrazole (PTZ, 80 mg/kg bw) induced convulsant. Molecular docking study predicted good total score of the swertiamarin with the selected targets. Swertiamarin possesses antiproliferative activity on HEp-2 and HT-29 cells with lower CTC50 values. It also served as significant anticonvulsant agent with prolonged onset and reduced duration of the seizures. These results confirm that swertiamarin exhibited potential antiproliferative and anticonvulsant activities.

[A new naphthaldehyde derivative from Comastoma pulmonarium and its anti-tobacco mosaic virus (anti-TMV) activity].

A new naphthaldehyde derivative has been isolated from Comastoma pulmonarium by using various chromatographic techniques, including silica gel, Sephadex LH-20, MCI-gel resin and RP-HPLC. This compounds was determined as 5-methoxy-2-methyl-7-(2-oxopropyl)naphthalene-1-carbaldehyde(1) by NMR, MS, IR and UV spectra. This compound was also evaluated for its anti-tobacco mosaic virus (anti-TMV) activity. The result showed that it showed high anti-TMV activity with inhibition rate of 32.8%. The inhibition rate is close to that of positive control (ningnanmycin).

Swertia mussotii extracts induce mitochondria-dependent apoptosis in gastric cancer cells.

Swertia mussotii (Gentianaceae) is a traditional Chinese medicinal plant grown in the Qinghai-Tibet Plateau. Three fractions from S. mussotii extract, named SWF50, SWF 70 and SWF100, were screened for in vitro anti-proliferative activity on two gastric cancer cell lines, MGC-803 and BGC-823 cells using MTT assay. Our results demonstrated that SMF70 showed an anti-proliferative effect in MGC-803 cells and SMF100 showed an anti-proliferative effect in BGC-823 cells in vitro. Moreover, both two fractions induced apoptosis via depolymerization of cytoskeletal filaments, increased cytoplasmic levels of ROS and Ca2+ and disrupted mitochondrial transmembrane potential. In addition, flow cytometry analysis indicated that both two fractions could induce cell apoptosis and arrest the cell cycle at S phase. Our results indicate that SMF70 induces apoptosis of MGC-803 cells and SMF100 induces apoptosis of BGC-823 cells via a mitochondrial-dependent pathway. Meanwhile, we also investigated antitumor effect of SMF70 in vivo, and exhibited effective tumor growth inhibition. Our findings demonstrate that S. mussotii extracts could be a potential new alternative therapeutic agent gastric cancer.

Hepatoprotective activity of iridoids, seco-iridoids and analog glycosides from Gentianaceae on HepG2 cells via CYP3A4 induction and mitochondrial pathway.

Gentianaceae herb extracts have been widely used as food additives, teas or medicinal remedies for various diseases and disorders of the human body. Herein, the potential effects of iridoids, seco-iridoids and analog glycosides from gentian on acontine-induced hepatotoxicity were investigated in HepG2 cells to obtain metabolic data of drug-biotarget interactions. Molecular docking analysis was performed to assess the binding efficiencies of 53 iridoids, seco-iridoids and analog compounds obtained from 50 gentian species to the active sites of human CYP3A4 enzyme. The docking scores of 29 iridoids, seco-iridoids and 24 analog glycosides were calculated from the free energy of ligand-protein complexes using a computer-assisted docking simulation. After comprehensive evaluation, 6 of these compounds, i.e., gentiopicroside, sweroside, swertiamarin, loganic acid, 6-O-ß-d-glucosyl-gentiopicroside and amarogentin were selected to evaluate their hepatoprotective effects. Quantitative real-time PCR was used to measure the expression levels of CYP3A4 mRNA in HepG2 cells. Amarogentin displayed the most clear inductive effect on CYP3A4 mRNA levels in the HepG2 cells. Moreover, amarogentin was further studied for acontine-induced toxicity in the HepG2 cells to determine the potential mechanisms. Amarogentin displayed obvious inductive effect on CYP3A4 mRNA levels in the HepG2 cells. These results elucidated that the hepatoprotective effects were caused by the facilitation of drug metabolism, amelioration of mitochondrial dysfunction and reduction of oxidative stress. Our data demonstrated that the naturally found iridoids, seco-iridoids and analog glycosides in gentian may be responsible for the hepatoprotective effects of gentian-extracted compounds and thus, this study may be useful in the food industry or in clinical practice.

Roburic Acid Suppresses NO and IL-6 Production via Targeting NF-κB and MAPK Pathway in RAW264.7 Cells.

In the present study, we investigated the anti-inflammatory effect of roburic acid on production of nitric oxide (NO) and interlukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. We found that roburic acid reduced production of NO and IL-6, and the expression of inducible nitric oxide synthases (iNOS). Meanwhile, phosphorylation of inhibitor of κBα (IκBα) and IκB kinase α/ß (IKKα/ß), as well as translocation of nuclear factor-κB (NF-κB) to the nucleus, was suppressed by roburic acid treatment. In addition, phosphorylation of mitogen-activated protein kinase (MAPKs) including p38 and c-Jun-NH2-terminal kinase (JNK) was inhibited. Roburic acid exhibited inhibitory activities on production of NO and IL-6 via blocking IKK/IκB/NF-κB and MAPKs pathway, suggesting the potential application as a drug candidate for therapy of inflammatory diseases.

Flavonoids from Enicostema littorale blume enhances glucose uptake of cells in insulin resistant human liver cancer (HepG2) cell line via IRS-1/PI3K/Akt pathway.

Diabetes mellitus has spread over the world with 347 million people affected. Insulin resistance is a main pathogenic event in Type 2 Diabetes Mellitus (T2DM) leading to a reduction in glucose uptake by peripheral tissue and increased hepatic glucose output. In this study, we have isolated four flavonoid rich fractions fraction A (FA), fraction B (FB), fraction C (FC) and fraction D (FD) from Enicostema littorale. All the fractions were preliminary screened for TLC fingerprinting, total flavonoid content. Total eight flavonoids were identified by LC/MS. Insulin resistant HepG2 (IR/HepG2) model was established by inducing insulin resistance in HepG2 cells to investigate the effect of these fractions on IR/HepG2 cell line for their glucose uptake. The results showed the significant dose dependant increase in glucose uptake of cells treated with FD. It showed significant activity at a concentration of 10µg/ml. The LC/MS results of FD demonstrated the presence of C-glycoside Swertisin which could be responsible for the effect. Further, to investigate the mechanism of action, gene expression for insulin receptor substrate 1 (IRS-1), protein kinase B (Akt-2) and glucose transporter 4 (GLUT-4) genes were evaluated by real time PCR. A significant upregulation of these genes was observed in FD treated samples, thereby indicating the enhancement of glucose uptake rate of cells via IRS-1/PI3K/Akt pathway.

The water extract of Veratrilla baillonii could attenuate the subacute toxicity induced by Aconitum brachypodum.

BACKGROUND: Aconitum brachypodum Diels (Family Ranunculaceae) is a Chinese ethnodrug and is well known for both its therapeutic application and high toxicity. However, no detoxication strategy is available for the complete elimination of the toxicity of Aconitum plants. Veratrilla baillonii Franch is believed to possess antitoxic effects on the toxicity induced by Aconitum plants and has been clinically used for hundreds of time by Naxi and Lisu nationalities in Yunnan Province of China. To further address the mechanism of the detoxication of Veratrilla baillonii, the effect of water decoction of Veratrilla baillonii (WVBF) on subacute toxicology of SD rats induced by Aconitum brachypodum (CFA), a genus Aconitum, was determined and studied in the present work. METHODS: The clinical behavior and number of survivors for different dosage of WVBF (25, 50, 100mg/kg) on CFA (4mg/kg) induced rats were observed until day 28. Histological changes and haematological parameters were evaluated. Moreover, Na+-K+-ATPase pathway in heart as well as key enzymes in liver were determined to further discuss the mechanism. RESULTS: The results showed that the exposure of CFA led to some subacute toxicity to rats, especially male ones, accompanied with abnormality of serum biochemical index in rats' serum. The toxicological target organs of CFA may be the heart, liver, kidney and brain. It is demonstrated that WVBF could attenuate the toxicity induced by Aconitum brachypodum via promoting the metabolic enzymes CYP3A1 and CYP3A2 in liver, downregulating the expression of Sodium/Calcium exchanger 1 (NCX1) and SCN5A sodium channal mRNA, and inducing Na+/K+-ATPase activity in heart. This study provides insights into detoxifying measures of Aconitum plants. CONCLUSIONS: Aconitum brachypodum may lead to subacute toxicity of rats after long term of administration, and the toxicity could be attenuated by Veratrilla baillonii via promoting the metabolic enzymes in liver, downregulating the expression of NCX1 and SCN5A mRNA, and inducing Na+/K+-ATPase activity in heart.

Mangiferin and Cancer: Mechanisms of Action.

Mangiferin, a bioactive compound derived primarily from Anacardiaceae and Gentianaceae families and found in mangoes and honeybush tea, has been extensively studied for its therapeutic properties. Mangiferin has shown promising chemotherapeutic and chemopreventative potential. This review focuses on the effect of mangiferin on: (1) inflammation, with respect to NFκB, PPARÒ¯ and the immune system; (2) cell cycle, the MAPK pathway G2/M checkpoint; (3) proliferation and metastasis, and implications on ß-catenin, MMPs, EMT, angiogenesis and tumour volume; (4) apoptosis, with a focus on Bax/Bcl ratios, intrinsic/extrinsic apoptotic pathways and telomerase activity; (5) oxidative stress, through Nrf2/ARE signalling, ROS elimination and catalase activity; and (6) efficacy of chemotherapeutic agents, such as oxaliplatin, etoposide and doxorubicin. In addition, the need to enhance the bioavailability and delivery of mangiferin are briefly addressed, as well as the potential for toxicity.

Medicinal plants of the genus Anthocleista--A review of their ethnobotany, phytochemistry and pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: The genus Anthocleista of the Gentianaceae family contains 14 species of trees and shrub-like plants distributed in tropical Africa, in Madagascar and on the Comoros. Traditionally, they are commonly used in the treatment of diabetes, hypertension, malaria, typhoid fever, obesity, diarrhea, dysentery, hyperprolactinemia, abdominal pain, ulcer, jaundice, asthma, hemorrhoids, hernia, cancer, wounds, chest pains, inflammations, rheumatism, STDs, infertility and skin diseases. They serve as an anthelmintic, laxative, diuretic and contraceptive. This review aims to provide for the first time a repository of ethnopharmacological information while critically evaluating the relation between the traditional medicinal uses, chemical constituents and pharmacological activities of the Anthocleista species so as to unveil opportunities for future research. MATERIALS AND METHODS: A search for relevant information on Anthocleista species was performed on scientific databases (Pubmed, Google Scholar, SciFinder, Web of Science, Scopus, PubChem and other web sources such as The Plant List, Kew Botanical Garden and PROTA) and books, PhD and MSc dissertations for un-published resources. RESULTS: Out of the 14 species of Anthocleista, 6 have been reported in literature to be widely used in traditional medicine for the treatment of various ailments. The six species include: A. djalonensis, A. vogelii, A. nobilis, A. grandiflora, A. schweinfurthii, and A. liebrechtsiana. The chemical compounds isolated from Anthocleista species fall into the class of phytochemicals such as secoiridoids, nor-secoiridoids, xanthones, phytosterols, triterpenes, alkaloids, and others of which majority of the compounds were isolated from A. djalonensis and A. vogelii. The in vitro and in vivo pharmacological studies on the crude extracts, fractions and few isolated compounds of Anthocleista species showed antidiabetic, antiplasmodial, antimicrobial, hypotensive, spasmogenic, anti-obesity, antiulcerogenic, analgesic, anti-inflammatory, antioxidant, antitrypanosomal, anthelmintic, fertility, diuretic and laxative activities which supports most of their uses in traditional medicine. However, the bulk of the studies where centered on the antidiabetic, antiplasmodial and antimicrobial activities of Anthocleista species, although the evidence of its antiplasmodial effect was not convincing enough due to the discrepancies between the in vitro and in vivo results. CONCLUSION: A. djalonensis and A. vogelii are potential antidiabetic and antibacterial agents. The antibacterial potency relates to infections or diseases caused by E. coli, S. typhi and S. aureus such as urinary tract infections, typhoid, diarrhea, skin diseases, and food poisoning. Pharmacological research on this genus is quite elementary and limited, thus, more advanced research is necessary to isolate and determine the activities of bioactive compounds in vitro and in vivo, establish their mechanisms of action and commence the process of clinical research.

Enicostemma littorale prevents tumor formation in 7,12-dimethylbenz(a)anthracene-induced hamster buccal pouch carcinogenesis.

Oral cancer is one of the most common malignancies worldwide, and India has recorded the highest annual incidence of oral cancer in comparison with other countries. Altered lipid peroxidation and antioxidant status along with defect in detoxification cascade have been implicated in the pathogenesis of several cancers including oral cancer. The aim of this study was to investigate the chemopreventive potential of ethanolic extract of Enicostemma littorale leaves (ElELet) in 7,12-dimethylbenz(a)anthracene (DMBA)-induced hamster buccal pouch carcinogenesis. Oral tumor was developed in the buccal pouches of male golden Syrian hamsters by painting with 0.5% DMBA three times a week for 14 weeks. We observed 100% tumor formation with increase in tumor volume and tumor burden in the hamsters treated with DMBA alone. Imbalance in phase I (cytochrome P450 and cytochrome b5) and phase II (glutathione reductase, glutathione-S-transferase, glutathione, and Deoxythymidine-diaphorase (DT)-diaphorase) detoxification agents and lipid peroxidation by-products (thiobarbituric acid reactive substances) and antioxidant (superoxide dismutase, catalase, glutathione peroxidase, and vitamins E and C) status was noticed in hamsters treated with DMBA alone. Oral administration of ElELet at a dose of 250 mg/kg body weight to hamsters treated with DMBA significantly prevented both precancerous and cancerous lesions in the oral cavity. ElELet modulated the status of phase I and II detoxification agents and antioxidants in favor of the suppression of oral carcinogenesis. This study thus suggests that E. littorale might have inhibited the oral carcinogenesis in DMBA-treated hamsters through its antioxidant potential. The present findings are also substantiated by histological studies during DMBA-induced oral carcinogenesis.

[Chemical Constituents in Petroleum Ether Extract of Mongolian Medicine Halenia corniculata].

OBJECTIVE: To study the chemical constituents of Mongolian medicine Halenia corniculata. METHODS: Positive phase and reversed phase silica gel, as well as Sephadex LH-20 methods were used to separate and purify. The structure of the isolated constituents was identified according to the NMR spectroscopy data and the literature data. RESULTS: Nine compounds were isolated from 95% ethanol extracts of petroleum ether part of Halenia corniculata and identified as: 1-hydroxy-2,3,4,6-tetramethoxyxanthone (1), 1-hydroxy-2,3, 5-trimethoxyxanthone (2) 1-hydroxy-3,7-dimethoxyxanthone (3), 1-hydroxy-3,5,6,7,8-pentamethoxyxanthone (4), 1-hydroxy-2,3,4, 7-tetramethoxyxanthone (5), 1-hydroxy-3,5-dimethoxyxanthone (6),1-hydroxy-2,3,4,5,7-pentamethoxyxanthone (7), palmitic acid (8) and ß-sitosterol (9). CONCLUSION: Compounds 3, 4 and 8 are isolated from this genus for the first time, Compound 1 is isolated from this plant for the first time.

Factorial design of essential oil extraction from Fagraea fragrans Roxb. flowers and evaluation of its biological activities for perfumery and cosmetic applications.

OBJECTIVE: To optimize the extraction yields of essential oil from Fagraea fragrans Roxb. flowers in hydro-distillation using a central composite design (CCD) and to evaluate its biological activities for perfumery and cosmetic applications. METHODS: Central composite design was applied to study the influences of operational parameters [water to flower weight (X(1)) and distillation time (X(2))] on the yields of essential oil (Y). Chemical compositions of the essential oil extracted from the optimized condition were identified by gas chromatography-mass spectrometry. Antioxidant activities of the essential oil were determined against ABTS(•+) and DPPH(•) radicals, and the cytotoxic effects were assessed on human embryonic kidney (HEK293) cells by the use of the MTT assay. Also, the aromatic properties of the essential oil were evaluated by five healthy trained volunteers. RESULTS: The best conditions to obtain the maximum essential oil yield were 7.5 mL g(-1) (X(1)) and 215 min (X(2)). The experimental yield of the essential oil (0.35 ± 0.02% v/w) was close to the value predicted by a mathematical model (0.35 ± 0.01% v/w). 3-Octadecyne, Z,Z,Z-7,10,13-hexadecatrienal, E-nerolidol, pentadecanal and linalool were the major constituents of the essential oil. The essential oil showed moderate antioxidant capacities with no toxic effects on HEK293 cells at 1-250 µg mL(-1). Also, the essential oil exhibited a very strong aroma and was classified to be top- to middle-notes. CONCLUSION: The results offer the effectively operational conditions in the extraction of essential oil from F. fragrans using hydro-distillation. The essential oil could be used as a natural fragrance, having antioxidant activity with slight cytotoxicity, for perfumery and cosmetic applications.

Hepatoprotective saikosaponin homologs from Comastoma pedunculatum.

Eight new triterpenoid saponins, the saikosaponin homologs comastomasaponins A-H (1-8), as well as a known triterpenoid (9) and eight known saponins (10-17) were isolated from the aerial portions of Comastoma pedunculatum. The structures of these compounds were elucidated spectroscopically, and their hepatoprotective activity and cytotoxic activity were evaluated against five human tumor cell lines in vitro. Compounds 1, 5-12, 14, 15, and 17 exhibited potent hepatoprotective activity, and compound 11 displayed cytotoxic activity against HCT-8, Bel-7402, BGC-825, A549, and A2780 human tumor cell lines.

Two new 2,3-seco-hopane triterpene derivatives from Megacodon stylophorus and their antiproliferative and antimicrobial activities.

Chemical investigation of the ethanol extract of the whole plant of Megacodon stylophorus led to the isolation and identification of two new seco-hopane triterpenoids, 2,3-seco-22(29)-hopene-2-carboxyl-3-aldehyde (1) and 2,3-seco-4(23),22(29)-hopene-2-carboxyl-3-aldehyde (2), along with 10 known compounds, 3-12. All the isolates were reported from this plant for the first time. The structures of compounds 1 and 2 were determined by detailed analysis of their spectral data including 1D and 2D NMR. In addition, compound 1 was further analyzed by X-ray crystallography. Compounds 1-3 were evaluated for their in vitro anti-proliferative activities on HeLa, MCF-7, and Hep-G2 tumor cell lines. Compound 2 was active against the three cell lines with IC50 values of 3.6, 7.5, and 13.6 µM, respectively, while compound 1 exhibited cytotoxicity on MCF-7 (IC50 14.0 µM) and HeLa (IC50 18.2 µM) cell lines. Antimicrobial activities of compounds 1-2 (minimum inhibitory concentration values in the range of 3.12-12.50 mg/mL) were also observed.

In vitro and in vivo antimalarial activity and cytotoxicity of extracts, fractions and a substance isolated from the Amazonian plant Tachia grandiflora (Gentianaceae)

Tachia sp. are used as antimalarials in the Amazon Region and in vivo antimalarial activity of a Tachia sp. has been previously reported. Tachia grandiflora Maguire and Weaver is an Amazonian antimalarial plant and herein its cytotoxicity and antimalarial activity were investigated. Spectral analysis of the tetraoxygenated xanthone decussatin and the iridoid aglyone amplexine isolated, respectively, from the chloroform fractions of root methanol and leaf ethanol extracts was performed. In vitro inhibition of the growth of Plasmodium falciparum Welch was evaluated using optical microscopy on blood smears. Crude extracts of leaves and roots were inactive in vitro. However, chloroform fractions of the root and leaf extracts [half-maximal inhibitory concentration (IC50) = 10.5 and 35.8 µg/mL, respectively] and amplexine (IC50= 7.1 µg/mL) were active in vitro. Extracts and fractions were not toxic to type MRC-5 human fibroblasts (IC50> 50 µg/mL). Water extracts of the roots of T. grandiflora administered by mouth were the most active extracts in the Peters 4-day suppression test in Plasmodium berghei-infected mice. At 500 mg/kg/day, these extracts exhibited 45-59% inhibition five to seven days after infection. T. grandiflora infusions, fractions and isolated substance have potential as antimalarials.

Two new secoiridoids from Chelonanthus alatus (Aubl) Pulle (Gentianaceae) / Dos nuevos secoiridoides de Chelonanthus alatus (Aubl) Pulle (Gentianaceae)

The species Chelananthus alatus is an herbaceous plant with known ethno botanical and medicinal properties used in control of fever, especially those produced by malaria. From dried leaves (1.11 Kg), the crude alcoholic extract was fractionated by liquid-liquid partition with different polarity solvents. From the sec-butyl alcohol soluble fraction, by successive application of chromatographic methods, four compounds type iridoid were isolated and identified by spectroscopic techniques. Compound 1 is a new secoiridoid which was identified as sweroside 7-isobutyryloxy, and it is reported here for the first time in the Gentianaceae family; the other secoiridoids which were isolated are known as vogeloside (2), dihydro-chelonanthoside (3) and sweroside (4); vogeloside was identified for the first time in this plant (C. alatus). From the isopropyl acetate extract, in conjunction with the sweroside 7- isobutyryloxy (1), chelonanthoside (5) and sweroside (4), were identified, along with the sweroside 7-isovaleryloxy-(6) as a new side chain isomeric ester of dihydrochelonanthoside (3) . This work presents the spectroscopic analysis of the new structures and some bioactivity data.

La especie Chelonanthus alatus (Gentianaceae) es una hierba de aplicaciones ethnobotánicas reconocidas en medicina tradicional, especialmente en el control de la fiebre producida por la malaria. De las hojas secas (1,11 Kg) se realizó el extracto crudo en alcohol etílico, el cual se fraccionó por partición líquido-líquido (L-L) con disolventes de diferente polaridad. De la fracción soluble en alcohol sec-butílico, se aislaron cuatro compuestos tipo seco-iridoide por aplicación sucesiva de diversos métodos cromatográficos los cuales se identificaron por técnicas espectroscópicas. El compuesto 1 es un nuevo secoiridoide identificado como de 7- isobutiriloxi-swerosido, y se reporta por primera vez en la familia Gentianaceae; los otros tres secoiridoides aislados se conocen como vogelósido (2), dihidrochelonanthosido (3) y swerósido (4); el vogelósido se identificó por primera vez en C. alatus. De la fracción soluble en acetato de isopropilo además del 7-isobutiriloxi-swerosido (1) y el swerosido se aislaron e identificaron, el chelonanthosido (5) y el isovaleriloxi-swerosido (6), el cual es un nuevo isómero del dihidrochelonanthosido. En este trabajo se presenta el análisis espectroscópico que llevó a la elucidación estructural de los compuestos novedosos y algunos datos de bioactividad.

[Removal of cadmium from extracts of Gentianae Radix et Rhizoma by gamma-mercaptopropyl-modified silica gel].

OBJECTIVE: To study the optimum process of removing cadmium irons from extracts of Gentianae Radix et Rhizoma with gamma-mercaptopropyl-modified silica gel (MPS) and assess its cadmium ion-removing property. METHOD: Static and dynamic adsorptions were adopted to detect the cadmium-removing rate. MPS' cadmium ion-removing property was assessed with such indicators as the cadmium-removing rate, the solid content and the HPLC fingerprint. RESULT: The process parameters of the static adsorption were as follows: 0.20 g x mL(-1) of concentration of extracts, 120 minutes of adsorption time and 15:1 between raw materials and MPS. The process parameters of the dynamic adsorption were as follows: 1:3.5 times between diameter and height, 0.20 g x mL(-1) of concentration of extracts, 0.9 mL x min(-1) of flow rate of the extracts and 50:1 between raw materials and MPS. Before and after the cadmium ion-removing process, the extracts showed no notable difference in solid content and HPLC fingerprint. CONCLUSION: gamma-mercaptopropyl-modified silica gel (MPS) can effectively remove cadmium ion from the extracts of Gentianae Radix et Rhizoma with an excellent cadmium ion-removing property.

New xanthone glycosides from Comastoma pedunculatum.

Five new xanthone glycosides, comastomasides A-E (1-5), were isolated from aqueous ethanol extracts of the aerial portions of Comastoma pedunculatum. The structures of these compounds were elucidated by spectroscopic analysis methods. Compounds 1-5 were evaluated for their hepatoprotective activity and cytotoxicity against four human cancer cell lines by in vitro assays. Among them, compounds 3 and 5 exhibited potent hepatoprotective activity. However, none of the compounds displayed cytotoxic activity.