猪毛蒿精油的杀虫作用研究

本文首次报道了猪毛蒿(Artenisia acoparia)精油的杀虫作用.经活性跟踪,确证茵陈二炔即为猪毛蒿精油中的活性成分.本文还报道了茵陈二炔的光活化特性及其对昆虫酯酶的影响.     

中国不同地区楝属植物中川楝素含量测定

应用高速液相色谱(HPLC)对我国楝属(Melia)植物主要分布区的川楝、苦楝的树皮、果实及叶片中川楝素含量进行了测定。结果表明,采自不同地区川棘、苦楝样本中川楝素的含量差别较大。树皮中的含量以贵州(沿河),陕西(西乡)采的苦楝及河南(扶沟)采的川楝较高,分别为5.160、4.697和4.223mg/g。果实中川楝素的含量都较低,苦楝果中仅含微量或测不出。杭州采的川楝果中川楝素含量却显著高于其它楝果样本,而且比其树皮的含量还高(分别为0.800和0.574mg/g)。所采的楝叶中在该测定条件下测不到川楝素存在。楝科中其它9种非楝属植物树皮中不含有川楝素。从整个测定结果可以看出,川楝素只存在于楝属植物树皮中,果中也有少量分布,川楝中川楝素的含量不一定高于苦楝。各样本中川楝素含量的差异是否与其分布区的地理环境、气候及土壤等因素有关,有必要作进一步研究。     

Subamolide E from Cinnamomum subavenium induces sub-G1 cell-cycle arrest and caspase-dependent apoptosis and reduces the migration ability of human melanoma cells

The aim of this work was to investigate the anticancer cytotoxic effects of natural compound subamolide E on the human skin cancer melanoma A375.S2 cells. Subamolide E was isolated from Cinnamomum subavenium and demonstrated cytotoxicities in the cell-growth assay at concentration ranges from 0 to 100 μM at 24 h. Propidium iodide staining and flow cytometry analyses were used to evaluate cell-cycle distribution and found that subamolide E caused DNA damage in the sub-G1 phase with a dose-dependent manner after 24 h of treatment. According to the western blot result, subamolide-E-treated cells with the increase of caspase-dependent apoptotic proteins induced related pathway mechanisms. Subamolide E also showed antimigratory activities of A375.S2 cells on the wound-healing assay. Finally, subamolide E demonstrated minor cytotoxicities to normal human skin cells (keratinocytes, melanocytes, and fibroblasts); therefore, it is a potential chemotherapeutic agent against skin melanoma.     

Preventive effects of taurine on development of hepatic steatosis induced by a high-fat/cholesterol dietary habit

Nonalcoholic fatty liver (NAFL) is also called hepatic steatosis and has become an emergent liver disease in developed and developing nations. This study was to exam the preventive effects of taurine (Tau) on the development of hepatic steatosis via a hamster model. Although hepatic steatosis of hamsters was induced by feeding a high-fat/cholesterol diet, drinking water containing 0.35 and 0.7% Tau improved (p < 0.05) the serum lipid profile. Meanwhile, the smaller (p < 0.05) liver sizes and lower (p < 0.05) hepatic lipids in high-fat/cholesterol dietary hamsters drinking Tau may be partially due to higher (p < 0.05) fecal cholesterol, triacylglycerol, and bile acid outputs. In the regulation of lipid homeostasis, drinking a Tau solution upregulated (p < 0.05) low-density lipoprotein receptor and CYP7A1 gene expressions in high-fat/cholesterol dietary hamsters, which result in increased fecal cholesterol and bile acid outputs. Drinking a Tau solution also upregulated (p < 0.05) peroxisome proliferator-activated receptor-α (PPAR-α) and uncoupling protein 2 (UPC2) gene expressions in high-fat/cholesterol dietary hamsters, thus increasing energy expenditure. Besides, Tau also enhanced (p < 0.05) liver antioxidant capacities (GSH, TEAC, SOD, and CAT) and decreased (p < 0.05) lipid peroxidation (MDA), which alleviated liver damage in the high-fat/cholesterol dietary hamsters. Therefore, Tau shows preventive effects on the development of hepatic steatosis induced by a high-fat/cholesterol dietary habit.     

Anti-inflammatory effects of supercritical carbon dioxide extract and its isolated carnosic acid from Rosmarinus officinalis leaves

Rosemary (Rosmarinus officinalis) leaves possess a variety of bioactivities. Previous studies have shown that the extract of rosemary leaves from supercritical fluid extraction inhibits the expression of inflammatory mediators with apparent dose-dependent responses. In this study, three different extraction conditions (5000 psi at 40, 60, and 80 °C) of supercritical carbon dioxide (SC-CO(2)) toward the extraction of antioxidants from rosemary were investigated. Furthermore, simultaneous comparison of the anti-inflammatory properties between rosemary extract prepared from SC-CO(2) under optimal conditions (5,000 psi and 80 °C) and its purified carnosic acid (CA) using lipopolysaccharide (LPS)-treated murine RAW 264.7 macrophage cells was also presented. Results showed that the yield of 3.92% and total phenolics of 213.5 mg/g extract obtained from the most effective extraction conditions showed a high inhibitory effect on lipid peroxidation (IC(50) 33.4 μg/mL). Both the SC-CO(2) extract and CA markedly suppressed the LPS-induced production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α), as well as the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), phosphorylated inhibitor-kappaB (P-IκB), and nuclear factor-kappaB (NF-κB)/p65 in a dose-dependent manner. The five major compounds of verbenone, cirsimaritin, salvigenin, carnosol, and CA existing in the SC-CO(2) extract were isolated by semipreparative HPLC and identified by HPLC-MS/MS analysis. CA was the most abundant recorded compound and the most important photochemical with an anti-inflammatory effect with an IC(50) of 22.5 μM or 7.47 μg/mL presented to the best inhibitory activity on NO production better than that of the 14.50 μg/mL dosage prepared from the SC-CO(2) extract. Nevertheless, the effective inhibition of LPS-induced NF-κB signaling in RAW 264.7 cells from the SC-CO(2) extract extends the potential application of nutraceutical formulation for the prevention of inflammatory diseases.     

Diminishing returns epistasis among beneficial mutations decelerates adaptation

Epistasis has substantial impacts on evolution, in particular, the rate of adaptation. We generated combinations of beneficial mutations that arose in a lineage during rapid adaptation of a bacterium whose growth depended on a newly introduced metabolic pathway. The proportional selective benefit for three of the four loci consistently decreased when they were introduced onto more fit backgrounds. These three alleles all reduced morphological defects caused by expression of the foreign pathway. A simple theoretical model segregating the apparent contribution of individual alleles to benefits and costs effectively predicted the interactions between them. These results provide the first evidence that patterns of epistasis may differ for within- and between-gene interactions during adaptation and that diminishing returns epistasis contributes to the consistent observation of decelerating fitness gains during adaptation.     

Tagging of Litopenaeus vannamei (Boone) by uropod cutting: a comparison of two methods

Two uropod‐cutting methods were used and compared as a means of long‐term tagging on juvenile (mean weight: 1.77±0.5 g) and sub‐adult (mean weight: 18.7±0.2 g) Litopenaeus vannamei: (a) cutting half of the left exopod (LE); and (b) cutting the whole right uropods (WR). Shrimp with uncut uropods served as controls. Regeneration of uropods in juvenile shrimp commenced within a week and complete regeneration was evident after 2 weeks. In sub‐adult shrimps, incomplete regeneration (IG) was observed within 1 month of culture in both cutting methods. Complete regeneration was observed in 2 months among shrimps in LE but not in WR. Abnormal growths (AGs) on regenerated uropods (91.1%) were observed in juvenile shrimp including hollow‐type (HT; 77.8%) and flat‐type (FT; 13.3%). Among sub‐adult shrimp, 87.6% produced AG including HT (68.8%) and FT (18.8%). In WR, regenerated uropods among juvenile shrimp were of a normal type (N) which was indistinguishable from the control treatment. On the other hand, normal growth (with less pigmentation, 5.2%), IG (26.3%), and AG (68.5%) were observed among sub‐adult shrimp. After 4 months of culture, with average final weights of 18–19 g, tag retention rate in juvenile shrimps was 91.1% for LE treatment, and 87.6% (LE) and 100% (WR) in sub‐adults. Survival rate was about 98% for both cutting methods in juvenile and 53–63% for sub‐adult.     

Proteomic Investigation of the Sinulariolide-Treated Melanoma Cells A375: Effects on the Cell Apoptosis through Mitochondrial-Related Pathway and Activation of Caspase Cascade

Sinulariolide is an active compound isolated from the cultured soft coral Sinularia flexibilis. In this study, we investigated the effects of sinulariolide on A375 melanoma cell growth and protein expression. Sinulariolide suppressed the proliferation and migration of melanoma cells in a concentration-dependent manner and was found to induce both early and late apoptosis by flow cytometric analysis. Comparative proteomic analysis was conducted to investigate the effects of sinulariolide at the molecular level by comparison between the protein profiles of melanoma cells treated with sinulariolide and those without treatment. Two-dimensional gel electrophoresis (2-DE) master maps of control and treated A375 cells were generated by analysis with PDQuest software. Comparison between these maps showed up- and downregulation of 21 proteins, seven of which were upregulated and 14 were downregulated. The proteomics studies described here identify some proteins that are involved in mitochondrial dysfunction and apoptosis-associated proteins, including heat shock protein 60, heat shock protein beta-1, ubiquinol cytochrome c reductase complex core protein 1, isocitrate dehydrogenase (NAD) subunit alpha (down-regulated), and prohibitin (up-regulated), in A375 melanoma cells exposed to sinulariolide. Sinulariolide-induced apoptosis is relevant to mitochondrial-mediated apoptosis via caspase-dependent pathways, elucidated by the loss of mitochondrial membrane potential, release of cytochrome c, and activation of Bax, Bad and caspase-3/-9, as well as suppression of p-Bad, Bcl-xL and Bcl-2. Taken together, our results show that sinulariolide-induced apoptosis might be related to activation of the caspase cascade and mitochondria dysfunction pathways. Our results suggest that sinulariolide merits further evaluation as a chemotherapeutic agent for human melanoma.     

Induction of Apoptosis by 11-Dehydrosinulariolide via Mitochondrial Dysregulation and ER Stress Pathways in Human Melanoma Cells

In this study the isolated compound 11-dehydrosinulariolide from soft coral Sinularia leptoclados possessed anti-proliferative, anti-migratory and apoptosis-inducing activities against A2058 melanoma cells. Anti-tumor effects of 11-dehydrosinulariolide were determined by MTT assay, cell migration assay and flow cytometry. Growth and migration of melanoma cells were dose-dependently inhibited by 2–8 μg/mL 11-dehydrosinulariolide. Flow cytometric data indicated that 11-dehydrosinulariolide induces both early and late apoptosis in melanoma cells. It was found that the apoptosis induced by 11-dehydrosinulariolide is relevant to mitochondrial-mediated apoptosis via caspase-dependent pathways, elucidated by loss of mitochondrial membrane potential (∆Ψm), release of cytochrome C, activation of caspase-3/-9 and Bax as well as suppression of Bcl-2/Bcl-xL. The cleavage of PARP-1 suggested partial involvement of caspase-independent pathways. Immunoblotting data displayed up-regulations of PERK/eIF2α/ATF4/CHOP and ATF6/CHOP coupling with elevation of ER stress chaperones GRP78, GRP94, calnexin, calreticulin and PDI, implicating the involvement of these factors in ER stress-mediated apoptosis induced by 11-dehydrosinulariolide. The abolishment of apoptotic events after pre-treatment with salubrinal indicated that ER stress-mediated apoptosis is also induced by 11-dehydrosinulariolide against melanoma cells. The data in this study suggest that 11-dehydrosinulariolide potentially induces apoptosis against melanoma cells via mitochondrial dysregulation and ER stress pathways.