Comparative evaluation of four Lycium barbarum cultivars on NaIO3-induced retinal degeneration mice via multivariate statistical analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruit of Lycium barbarum L. (goji berry) is a traditional Chinese medicine and is often used to improve vision. While various goji cultivars may differentially treat retinal degeneration, however their comparative effectiveness remains unclear. AIM OF THE STUDY: To evaluate the protective effects of four goji cultivars on NaIO3-induced retinal degeneration mouse model and identify the most therapeutically potent cultivar. MATERIALS AND METHODS: The principal compounds in the extracts of four goji cultivars were characterized by UPLC-Q-TOF/MS. A retinal degeneration mouse model was established via NaIO3 injection. Dark-light transition and TUNEL assays were used to assess visual function and retinal apoptosis. The levels of antioxidative, inflammatory, and angiogenic markers in serums and eyeballs were measured. Hierarchical cluster analysis, principal component analysis and partial least squares-discriminant analysis were used to objectively compare the treatment responses. RESULTS: Sixteen compounds were identified in goji berry extracts. All goji berry extracts could reverse NaIO3-induced visual impairment, retinal damage and apoptosis. The samples from the cultivar of Ningqi No.1 significantly modulated oxidative stress, inflammation, and vascular endothelial growth factor levels, which are more effectively than the other cultivars based on integrated multivariate profiling. CONCLUSION: Ningqi No.1 demonstrated a stronger protective effect on mouse retina than other goji cultivars, and is a potential variety for further research on the treatment of retinal degeneration.

Comparative Analysis of Chemical Composition andAntibacterial and Anti-Inflammatory Activities of theEssential Oils from Chrysanthemum morifolium ofDifferent Flowering Stages and Different Parts.

The inflorescence of Chrysanthemum morifolium Ramat., a well-known traditional Chinese herb, has been proved to have a certain inhibitory effect on some bacteria; however, its main components and acne bacteria inhibition effect remain to be elucidated. In this study, GC-MS was used to analyze the components of different flowering stages and different parts and to study the inhibitory effects of six essential oils on S. aureus and P. acnes and their alleviating effects on THP-1 cell inflammation. GC-MS combined with relative retention index method analyzed results stated that the 5 samples of C. morifolium to detect the 124 kinds of volatile components, including (E)-tibetin spiroether, are first detected in the volatile oil of the C. morifolium, and the content of (E)-tibetin spiroether is higher in immature inflorescence of C. morifolium and decreases as it extends its flowering period. Furthermore, the research results of inhibiting common acne-causing bacteria showed that the bacteriostatic effect of essential oils from JH at different flowering stages was better than that from JM and TJ, while the bacteriostatic effect of essential oil from stem and leaf of C. morifolium (SLC) at different parts was better than the roots of C. morifolium (RC). Finally, it was proved that the essential oil from SLC and C. morifolium could alleviate the inflammation of THP-1 cells induced by P. acnes. In conclusion, the antibacterial and anti-inflammatory effects of C. morifolium essential oil may be related to heterospiroolefins compounds, and the antibacterial activity decreases with the prolongation of flowering stage. It was suggested that volatile oil from C. morifolium and SLC could be used as effective components of antibacterial and anti-inflammatory cosmetics.

Tyrosine hydroxylase coordinates larval-pupal tanning and immunity in oriental fruit fly (Bactrocera dorsalis).

BACKGROUND: The oriental fruit fly Bactrocera dorsalis (Hendel), a notorious world pest infesting fruits and vegetables, has evolved a high level of resistance to many commonly used insecticides. In this study, we investigate whether tyrosine hydroxylase (TH) that is required for cuticle tanning (sclerotization and pigmentation) in many insects, could be a potential target in controlling B. dorsalis. RESULTS: We cloned TH cDNA (BdTH) of B. dorsalis. The complete open reading frame of BdTH (KY911196) was 1737 bp in length, encoding a protein of 578 amino acids. Quantitative real-time PCR confirmed that BdTH was highly expressed in the epidermis of 3rd instar larvae, and its expression increased prior to pupation, suggesting a role in larval-pupal cuticle tanning. When we injected dsBdTH or 3-iodo-tyrosine (3-IT) as a TH inhibitor or fed insect diet supplemented with 3-IT, there was significant impairment of larval-pupal cuticle tanning and a severe obstacle to eclosion in adults followed by death in most. Furthermore, injection of Escherichia coli into larvae fed 3-IT resulted in 92% mortality and the expressions of four antimicrobial peptide genes were significantly downregulated. CONCLUSION: These results suggest that BdTH might play a critical role in larval-pupal tanning and immunity of B. dorsalis, and could be used as a potential novel target for pest control. © 2017 Society of Chemical Industry.

Comparative pharmacokinetics of acteoside from total glycoside extracted from leaves of Rehmannia and Dihuangye total glycoside capsule in normal and diabetic nephropathy rats.

Rehmannia glutinosa Libosch (RG), is officially listed in the Chinese Pharmacopoeia and is widely used in China. In this paper, a sensitive and rapid ultra-performance liquid chromatography-mass spectrometry method including multiple-reaction monitoring mode was developed and applied to study the pharmacokinetic effect of acteoside from total glycoside extracted from the leaves of Rehmannia (TLR) and Dihuangye total glycoside capsule (DTG) in normal and diabetic nephropathy rats. The diabetic nephropathy rat model was induced by intraperitoneal injection of a small dose of streptozotocin and high-fat diet and plus 5% glucose drinking water. Samples of plasma of rats were obtained at different times after rats were administered TLR (7.2 g/kg) and DTG (360 mg/kg). After deproteinization by acetonitrile, the concentrations of acteoside in rats at different time points were detected by UPLC-TQ-MS method and pharmacokinetics parameters were calculated using DAS 3.2.8 software. A good linearity of acteoside was shown in the range of 8.51-3404.8 ng/m L (r2 = 0.9987). The mean extraction recovery of analyte was in the range of 63.55-79.49%, and the intra- and inter-day RSD values were <8.8%. Compared with the normal group, the maximum plasma concentration, AUC0-t , AUC0-∞ and apparent plasma clearance corresponding dose in model group rats decreased significantly. After rats were administered TLR and DTG, the acteoside reached the maximum plasma concentration at about 15 min. The method proved to be simple, rapid and specific, and to be suitable for the determination of acteoside in plasma of diabetic nephropathy rats and pharmacokinetic study.

(1)H NMR metabolomics to study the effects of diazepam on anisatin induced convulsive seizures.

The anticonvulsive properties of diazepam have been extensively studied, mainly focusing on the γ-amino butyrate (GABA) system. The aim of this investigation was to integrally analyze the metabolic events related to neuroprotection of diazepam on anisatin-induced convulsive seizures by a NMR-based metabolomic approach combined with histopathological examination and behavior examination. Multivariate analysis on metabolic profiles of the piriform cortex and cerebellum of mice revealed that diazepam could relieve mice suffering from the convulsive seizures by recovering destructed neurotransmitter and neuromodulator metabolism, ameliorating oxidative stress, alleviating the disturbance in energy, amino acid and nucleic acid metabolism in anisatin intoxicated mice. This integrated metabolomics study provided a powerful and highly effective approach to elucidate therapeutic effects and assessed the safety of diazepam. This study should be helpful for our understanding of convulsive seizures, and provide a holistic view of the treatment effects of benzodiazepine on convulsive seizures.

A bird's eye view of anisatin induced convulsive seizures in brain by a (1)H NMR based metabolic approach.

Anisatin is the main convulsant component in plants of the genus Illicium, many of which are important spices or folk medicines. The neurotoxicity of anisatin has been widely investigated, mainly focusing on its action on the γ-amino butyrate (GABA) system; however, little is known about the metabolic alterations that it causes. In this study, a NMR-based metabolomic approach was performed on the extracts of cortexes and cerebellums of mice administered with anisatin to explore the metabolic events associated with its intoxication. Orthogonal partial least squares-discriminant analysis (OPLS-DA) revealed many differential metabolites that indicated metabolic disturbance in neurotransmission and neuromodulation (GABA, glutamate, glutamine, and taurine), stress of reactive oxygen species (ROS) (ascorbate, phosphatidylcholine, choline, and ethanolamine), energy metabolism (NAD(+)i.e., nicotinamide-adenine dinucleotide, lactate, citrate, fumarate, creatine/phosphocreatine, and creatinine), amino acid metabolism (leucine, isoleucine, valine, phenylalanine, tyrosine, tryptophan, alanine, threonine, and glycine) and nucleic acid metabolism (NAD(+), nicotinamide/niacinamide, adenosine, and guanosine). This pilot metabolomic study on anisatin intoxication should help to develop a holistic view of convulsive seizures induced by anisatin, and provide a better understanding of the mechanisms.

Developmental toxicity and neurotoxicity of two matrine-type alkaloids, matrine and sophocarpine, in zebrafish (Danio rerio) embryos/larvae.

Matrine and sophocarpine are two major matrine-type alkaloids included in the traditional Chinese medicine (TCM) Kushen (the root of Sophora flavescens Ait.). They have been widely used clinically in China, however with few reports concerning their potential toxicities. This study investigated the developmental toxicity and neurotoxicity of matrine and sophocarpine on zebrafish embryos/larvae from 0 to 96/120h post fertilization (hpf). Both drugs displayed teratogenic and lethal effects with the EC50 and LC50 values at 145 and 240mg/L for matrine and 87.1 and 166mg/L for sophocarpine, respectively. Exposure of matrine and sophocarpine significantly altered spontaneous movement and inhibited swimming performance at concentrations below those causing lethality and malformations, indicating a neurotoxic potential of both drugs. The results are in agreement with most mammalian studies and clinical observations.

Bioactive terpenoids from the fruits of Aphanamixis grandifolia.

From the fruits of the tropical tree Aphanamixis grandifolia, five new evodulone-type limonoids, aphanalides I-M (1-5), one new apo-tirucallane-type triterpenoid, polystanin E (6), and three new chain-like diterpenoids, nemoralisins A-C (7-9), along with 12 known compounds were identified. The absolute configurations were determined by a combination of single-crystal X-ray diffraction studies, Mo2(OAc)4-induced electronic circular dichroism (ECD) data, the Mosher ester method, and calculated ECD data. The cytotoxicities of all the isolates and the insecticidal activities of the limonoids were evaluated.

Simonols A and B, two novel sesqui-neolignans from the fruits of Illicium simonsii.

Two new sesqui-neolignans with novel conjugation way, simonol A (1), featuring a unique motif of a 5,5-dihydro-pyran with a hemiketal carbon, while simonol B (2) possessing two dihydronfuran rings in the same direction, were isolated from the ethanol extract of the fruits of Illicium simonii. Their structures were elucidated by spectroscopic methods including 1D and 2D NMR, HRESIMS, and calculation of electronic circular dichroism (ECD) using density functional theory (DFT). The two isolates were evaluated for their inhibitory activities against the growth of four lines of human cancer cells (NCI-H460, SMMC-7721, MCF-7, BGC-823): 1 showed strong activities comparable to 5-Fluorouracil, and 2 to a less content. In addition, plausible biosynthetic routes for the two compounds were also proposed.

Glaucogenin E, a new C21 steroid from Cynanchum stauntonii.

Glaucogenin E (1), a new C(21) steroid sapogenin, along with three known ones (2-4) were isolated from the rhizomes of Cynanchum stauntonii (Decne.) Schltr. ex Levl. Their structures were established mainly by the spectroscopic analysis, including 2D NMR. All the isolated compounds were evaluated for their cytotoxicity against human cancer cell lines HeLa, Bel-7402, SGC-7901 and BGC-823.

A new phenylpropanoid glycoside from the fruits of Illicium simonsii.

AIM: To study the constituents and bioactivity of Illicium simonsii. METHODS: The extracts of the fruits of Illicium simonsii were submitted to a combination of chromatographic materials, silica gel, ODS column chromatography and finally preparative HPLC to obtain eight compounds which were further evaluated for their cytotoxicity by MTT method. RESULTS: A new phenylpropanoid glycolside, 2, 4-dihydroxy-allylbenzene-2-O-ß-D-glucopyranoside (1), together with seven characteristic sesquiterpene lactones, oligandrumin B (2), oligandrumin D (3), anisatin (4), veranisatin D (5), pseudomajucin (6), 1α-hydroxy-3-deoxy-pseudoanisatin (7), 8α-hydroxy-10-deoxycyclomerrillianolide (8) were isolated. CONCLUSION: Compound 1 is new. Compounds 1, 2, 3, 5-8 were isolated from this plant for the first time. None of the isolates showed inhibitory effects on the growth of non-small cell lung tumor cell line A549.

Reversal effects of components from the fruits of Illicium simonsii on human Adriamycin-resistant MCF-7 and 5-fluorouracil-resistant Bel7402 cells.

Twenty-one compounds including seven characteristic sesquiterpene lactones were isolated from Illicium simonsii and screened in vitro for their potential to restore the sensitivity of Adriamycin (ADR) resistant breast cancer cells (MCF-7/ADR) and 5-fluorouracil-resistant hepatocellular carcinoma cells (Bel7402/5-FU) to ADR and 5-fluorouracil, respectively. These compounds were found to be non-toxic to a panel of tumour cell lines: human breast cancer cells (MCF-7), cervical cancer cells (HeLa), hepatic liver carcinoma cells (HepG-2, Bel7402) and gastric carcinoma cells (BGC-823, SGC-7901). Five compounds showed an obvious decrease in the IC(50) of doxorubicin in MCF-7/ADR and four compounds sensitized Bel7402/5-FU to 5-fluorouracil at non-toxic concentrations. The relationship between the structure of these non-cytotoxic substances and their multidrug-resistant (MDR) reversal abilities was investigated by principal component analysis (PCA) of the reversing fold (RF) values of these compounds and their calculated molecular descriptors of the tested substances. No correlations were found between the reversal potencies of these compounds to the two MDR cells. Compounds with lower polarity generally had stronger sensitizing ability to the P-glycoprotein (Pgp) overexpressed MCF-7/ADR cells. On the other hand, higher hydrophilic compounds seemed to exhibit a stronger reversal effect to the MDR-associated protein (MRP) overexpressed Bel7402/5-FU cell line. Our findings favoured further investigations on the active substances and the underlying mechanisms.

Bioactivity-guided isolation of antiproliferative diterpenoids from Euphorbia kansui.

Cytotoxic assay guided multistep separation on the dichloromethane extract of the roots of Euphorbia kansui resulted in the isolation of 10 ingenol-type diterpenoids (1-4 and 7-12), of which, 5-O-(2'E,4'E-decadienoyl)-20-O-acetylingenol (1) is a new compound, and two are jatrophane-type diterpenoids (13-14). Interconversion of two pairs of positional ester isomers (1-4) in aqueous alcoholic solution was observed, the transesterification mechanism of which was speculated and confirmed by acylation of 3 and 4 to 6 and 5, respectively. All the isolates and the two acyl derivatives (5 and 6) were evaluated in vitro for their cytotoxicities in Bel-7402, Bel-7402/5FU, BGC-823 and SGC-7901 cell lines. The 12 ingenol-type diterpenoids exhibited weak to moderate cytotoxicities, whereas the two jatrophane-type diterpenoids displayed no antiproliferative effects, which, however, may increase the antitumour efficacy of those ingenol-type diterpenoids. The structure--activity relationships were investigated by principal component analysis (PCA) of the pIC50 (-logIC50) values of the compounds tested and their calculated molecular descriptors. The pIC50 values were highly correlated with most descriptors, especially the highest occupied molecular orbital energy (E(HOMO)), absolute hardness (η) and positively charged solvent accessible surface areas (P-ASA). As the values of E(HOMO) increase, η and P-ASA decrease, and the antiproliferative effects of these compounds increase.

Reversal effects of traditional Chinese herbs on multidrug resistance in cancer cells.

Multidrug resistance (MDR) continues to be a major obstacle for successful anticancer therapy. In this work, fractions from 17 clinically used antitumour traditional Chinese medicinal herbs were tested for their potential to restore the sensitivity of MCF-7/ADR and A549/Taxol cells to a known antineoplastic agent. The effects of these fractions were evaluated by MTT method and an assay of the cellular accumulation of doxorubicin. Fractions from the PB group (herbs with the ability to promote blood circulation and remove blood stasis) showed more significant effects than fractions from the CH group (herbs with the ability to clear away heat and toxic materials). Fractions from CH2Cl2 extracts were more effective than fractions from EtOAc extracts. Five herbs (Curcuma wenyujin, Chrysanthemum indicum, Salvia chinensis, Ligusticum chuanxiong Hort. and Cassia tora L.) could sensitise these resistant cancer cells at a non-toxic concentration (10 µg mL⁻¹), and markedly increased doxorubicin accumulation in MCF-7/ADR cells, which necessitates further investigations on the active ingredients of these herbs and their underlying mechanisms.