Identification of bioactive compounds and inhibitory effects of TNF-α and COX-2 in the extract from cultured three-spot seahorse (H. trimaculatus).

Three-spot seahorse (Hippocampus trimaculatus) has been consumed as traditional Chinese medicine in Asian society. This study was designed to analyze the bioactive compounds of the solvent extracts from cultured three-spot seahorse by high pressure liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI/MS/MS). Subsequently, their biological activities were evaluated and confirmed by cell modes and Western blot analysis. Experimental results indicated that taurine and arginine were the primary bioactive compounds identified and quantified without pre- or post-column derivatization within 20 min retention time. The analytical method was established and validated with intraday/interday RSD from 0.25% to 3.34% and with recovery from 87.8% to 91.2%. As compared to other extracts, water layer extract (WLE) contained the most taurine and arginine contents of 6.807 and 0.437 mg/g (dry basis), respectively. In the meanwhile, WLE also showed anti-inflammatory activity on LPS-induced NO production and inhibited the protein expression of TNF-α and COX-2 by Western blot analysis with better cell viability.

Structural characterization and anti-inflammatory activity evaluation of chemical constituents in the extract of Trifolium repens L.

The chemical constituents in Trifolium repens L. were comprehensively studied by UPLC in this work, and a total number of 308 compounds were detected with 169 ones identified. The possible fragmentation pathways were proposed and fragmentation rules were summarized. On the basis of the concluded strategies, the characterized compounds could be classified into organic acids and their derivatives, alkaloids, amino acids, peptides, flavonoids, oligosaccharides, coumarins, and other types of compounds. This approach provided a rapid way for the identification of constituents in T. repens L., and even in other complex analytes. Among the separation and identification of the constituents, three compounds of great amount were isolated and characterized by NMR. The expression of iNOS and COX-2 in LPS-induced RAW 264.7 cells was suppressed by the pretreatment with three isolated constituents. The results implied they may potentially serve as a remedy for the therapy of inflammation. PRACTICAL APPLICATIONS: This work provided a rapid method for the identification of the complex analyte, which could be used in TCM, natural food and so on. The summarized fragmentation rule could be applied for the analysis of several types of compounds, such as organic acids and their derivatives, alkaloids, amino acids and peptides, flavonoids, oligosaccharides, coumarins, and so on. Most of natural plants contain these kinds of compounds, so these rules could have wide applications. Except the phytochemical investigation, T. repens L. displayed anti-inflammation activity according to the reported literature, and the three isolated constituents may potentially serve as a remedy for the therapy of inflammation referring to the result of this research.

Study on Chemical Profile and Neuroprotective Activity of Myrica rubra Leaf Extract.

The chemical profile of Myrica rubra (a native species in China) leaf extract was investigated by UPLC-PDA-HRMS, and the neuroprotective activity of two characteristic constituents, myricanol and myricetrin, was evaluated with N2a cells using H2O2-inducedoxidative challenge through a series of methods, e.g., MTT assay, ROS assay and [Ca2+]i assay. Among the 188 constituents detected in the extract of Myrica rubra leaf, 116 were identified definitely or tentatively by the comprehensive utilization of precise molecular weight and abundant multistage fragmentation information obtained by quadrupole orbitrap mass spectrometry. In addition, 14 potential new compounds were reported for the first time. This work established an example for the research of microconstituents in a complex analyte and revealed that suppression of H2O2-induced cytotoxicity in N2a cells was achieved by the pretreatment with myricanol. The evidence suggested myricanol may potentially serve as a remedy for prevention and therapy of neurodegenerative diseases induced by oxidative stress.

Cladieunicellin J, a new hydroperoxyeunicellin from cladiella sp.

A new 6-hydroperoxyeunicellin diterpenoid, designated as cladieunicellin J (1), was isolated from an octocoral Cladiella sp., and its structure elucidated by spectroscopic methods. Compound 1 was found to exhibit cytotoxicity toward CCRF-CEM human T-cell acute lymphoblastic leukemia.

Cladieunicellin H, a new hemiketal eunicellin-based diterpenoid from the octocoral Cladiella sp.

A new hemiketal eunicellin-based diterpenoid, cladieunicellin H (1), was isolated from an Indonesian octocoral identified as Cladiella sp. The structure of 1 was determined by spectroscopic methods. This compound was found to display moderate inhibitory effects on the generation of superoxide anion and the release of elastase by human neutrophils.

New cytotoxic lupane triterpenes from Perrottetia arisanensis.

Eight new lupane triterpenes, including 7beta-cis-coumaroylbetulinic acid (1), 7beta-trans-coumaroylbetulinic acid (2), 7beta-cis-coumaroyl-3-epi-betulinic acid (3), 7beta-trans-coumaroyl-3-epi-betulinic acid (4), 7beta-cis-coumaroylbetulonic acid (5), 7beta-trans-coumaroylbetulonic acid (6), 7beta-hydroxybetulinaldehyde (7) and 28-norlup-20(29)-ene-3alpha,17beta-diol (8), together with fifteen known compounds were isolated from the bioactive methanol extract of the stems of Perrottetia arisanensis. The structures of the new compounds were elucidated by spectroscopic and HR-ESI-MS analysis. All new compounds were evaluated for their cytotoxicity against six human cancer cell lines. Among them, lupane triterpene coumaroyl esters 1-6 showed moderate cytotoxicity with IC (50) values ranging from 3.75 to 21.29 microM. This is the first report for lupane triterpenes with a phenylpropane moiety substituted at C-7.

Antioxidant activities of extracts and main components of Pigeonpea [Cajanus cajan (L.) Millsp.] leaves.

Antioxidant activities of the aqueous and ethanol extracts of pigeonpea [Cajanus cajan (L.) Millsp.] leaves, as well as petroleum ether, ethyl acetate, n-butanol and water fractions and the four main compounds separated from the ethanol extract, i.e. cajaninstilbene acid (3-hydroxy-4-prenylmethoxystilbene-2-carboxylic acid), pinostrobin, vitexin and orientin, were examined by a DPPH radical-scavenging assay and a beta-carotene-linoleic acid test. In the DPPH system, the antioxidant activity of the ethanol extracts was superior to that of the aqueous extracts, with IC(50) values were 242.01 and 404.91 microg/mL, respectively. Among the four fractions, the ethyl acetate one showed the highest scavenging activity, with an IC(50) value of 194.98 microg/mL. Cajaninstilbene acid (302.12 microg/mL) and orientin (316.21 microg/mL) showed more efficient radical-scavenging abilities than pinostrobin and vitexin. In the beta-carotene-linoleic acid test, the inhibition ratio (%) of the ethyl acetate fraction (94.13%+/-3.41%) was found to be the highest, being almost equal to the inhibition capacity of the positive control BHT (93.89%+/-1.45%) at 4 mg/mL. Pinostrobin (>500 microg/mL) and vitexin (>500 microg/mL) showed insignificant antioxidant activities compared with cajaninstilbene (321.53 microg/mL) and orientin (444.61 microg/mL). In general, the ethyl acetate fraction of the ethanol extract showed greater activity than the main compounds in both systems, such results might be attributed to the synergistic effects of the components. The antioxidant activities of all the tested samples were concentration-dependent. Based on the results obtained, we can conclude that the pigeonpea leaf extracts may be valuable natural antioxidant sources and are potentially applicable in both medicine and the healthy food industry.

New cytotoxic 6-oxygenated 8,9-dihydrofurocoumarins, hedyotiscone A - C, from Hedyotis biflora.

Using the bioactivity-guided fractionation method, three new 6-oxygenated 8,9-dihydrofurocoumarin-type compounds, hedyotiscones A, B, and C were isolated from the methanol extract of Hedyotis biflora together with seven known compounds. The structures of all isolated compounds were determined on the basis of mass and spectroscopic evidence. Compounds, oleanolic acid, and 6'-(beta-sitosteryl-3- O-beta- D-glucopyranosidyl) pentadecanoate showed marginal cytotoxicity against Hep G2 cells (human liver cancer cells) with IC50 values of 14.4, 17.4, 4.9, 8.0, and 9.2 microg/mL, respectively. Ursolic acid showed significant cytotoxicity against MDA-MB-231 cells (human breast carcinoma cells) with an IC50 value of 1.49 microg/mL.