An acidic polysaccharide, H2, was isolated from the alkali-extract CHC of seeds of Cuscuta chinensis Lam. with the molecular weight more than 1.0 x 10(6). Chemical and spectroscopic studies led to the structure determ...An acidic polysaccharide, H2, was isolated from the alkali-extract CHC of seeds of Cuscuta chinensis Lam. with the molecular weight more than 1.0 x 10(6). Chemical and spectroscopic studies led to the structure determination as follows: the backbone chain consists of 1, 6-linked-beta -D Galp, 1,4-linked-X -D Galp, 1,4-linked-beta -D GalA and 1,2- or 1,4-linked-beta -L Rhap having branching points at position O-3 of some 1,6-linked-beta -D Galp residues (one among eight) and O-4 or O-2 of 1,2- or 1,4-linked-beta -L Rhap residues to terminal beta -D-galactopyranose. The side chains composed of terminal Galp, 1,6-linked-beta -D Galp, 1,4-linked beta -D Galp and 1,3,6-linked-beta -D Galp also linked at position O-3 of 1,6-linked-beta -D Galp residues in the backbone chain. beta -L-arabinofuranosyl and terminal beta -L-rhamnopyranosyl residues existed in the periphery of this polysaccharide linked to O-3 of 1,6-linked-beta -D Galp residues in the backbone chain and the side chains. The polysaccharide H2 increased significantly the survival rate of PC12 cells indicating that it had protective effects against H2O2 insult.展开更多
Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW...Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW264.7) were stimulated by LPS alone, or with long-term of PMA pretreatment. Then cytotoxicities to P815 cells (by MTT assay) and IL-1, TNF- (by ELISA) and nitric oxide (NO) production (by Griess reagent) in supernatants were measured. Western blot for PKC isoforms after long-term PMA pretreatment was analyzed. Results: RAW264.7 cells were stimulated with LPS to kill target tumor cells P815, whereas P388D1 cells failed to develop such an ability. Down-regulation of PKC isoforms by chronic treatment with PMA significantly inhibited the LPS-induced cytotoxicity in RAW264.7 cells. In unstimulated state, Western blotting with rabbit antiserum specific for the PKC, 1, 2, or showed all 5 isoforms were detected in P388D1 cells, while only PKC, PKC1 and PKC were detected in RAW264.7 cells. Exposure of the cells to long-term of PMA treatment significantly down-regulated the expression of PKC, PKC1 and PKC in RAW264.7 cells. But in P388D1 cells, although PKC, PKC and PKC were down-regulated, the expression of PKC1 and PKC2 could not be regulated. Comparing with LPS-induced IL-1, TNF- and NO production by the two macrophage cell lines, P388D1 failed to produce NO. In RAW264.7 cells, LPS-induced NO production and antitumor activity was attenuated by the addition of L-NAME, an iNOS inhibitor. Conclusion: The results indicated a critical role of PKC in LPS-induced antitumor activity and this cytotoxicity is mainly due to PKC- mediated NO production by RAW264.7 cells, but not a direct cytotoxic activity.展开更多
AIM:To investigate the antioxidant activity of chitooligosaccharides(COSs)on pancreatic islet cells in diabetic rats induced by streptozotocin. METHODS:The antioxidant effect of COSs on pancreatic islet cells was dete...AIM:To investigate the antioxidant activity of chitooligosaccharides(COSs)on pancreatic islet cells in diabetic rats induced by streptozotocin. METHODS:The antioxidant effect of COSs on pancreatic islet cells was detected under optical microscopy and with colorimetric assay and gel electrophoresis.The activities of glutathione peroxidase and superoxide dismutase,total antioxidant capacity,and content of malondialdehyde in serum and tissue slices of pancreas were examined after 60 d to determine the effect of COSs in streptozotocin-induced diabetes in rats. RESULTS:COSs can prohibit the apoptosis of pancreatic islet cells.All concentrations of COSs can improve the capability of total antioxidant capacity and activity of superoxide dismutase and decrease the content of malondialdehyde drastically.Morphological investigation in the pancreas showed that COSs have resulted in the reduction of islets,loss of pancreatic cells,and nuclear pyknosis of pancreatic cells. CONCLUSION:COSs possess various biological activities and can be used in the treatment of diabetes mellitus.展开更多
Drug-induced liver injury is a significant and still unresolved clinical problem. Limitations to knowledge about the mechanisms of toxicity render incomplete the detection of hepatotoxic potential during preclinical d...Drug-induced liver injury is a significant and still unresolved clinical problem. Limitations to knowledge about the mechanisms of toxicity render incomplete the detection of hepatotoxic potential during preclinical development. Several xenobiotics are lipophilic substances and their transformation into hydrophilic compounds by the cytochrome P-450 system results in production of toxic metabolites. Aging, preexisting liver disease, enzyme induction or inhibition, genetic variances, local 02 supply and, above all, the intrinsic molecular properties of the drug may affect this process. Necrotic death follows antioxidant consumption and oxidation of intracellular proteins, which determine increased permeability of mitochondrial membranes, loss of potential, decreased ATP synthesis, inhibition of Ca^2+-dependent ATPase, reduced capability to sequester Ca^2+ within mitochondria, and membrane bleb formation. Conversely, activation of nucleases and energetic participation of mitochondria are the main intracellular mechanisms that lead to apoptosis. Non-parenchymal hepatic cells are inducers of hepatocellular injury and targets for damage. Activation of the immune system promotes idiosyncratic reactions that result in hepatic necrosis or cholestasis, in which different HLA genotypes might play a major role. This review focuses on current knowledge of the mechanisms of drug-induced liver injury and recent advances on newly discovered mechanisms of liver damage. Future perspectives including new frontiers for research are discussed.展开更多
The cytotoxicity of abamectin to the Gill Cell Line of Flounder (FG cell line) was examined in this study. It was found that the exposure of FG cells to abamectin caused the decreases of both cell growth rate and an...The cytotoxicity of abamectin to the Gill Cell Line of Flounder (FG cell line) was examined in this study. It was found that the exposure of FG cells to abamectin caused the decreases of both cell growth rate and antioxidant enzyme activities, and the increase of intracellular 02 content. It was proposed that the reduction of antioxidant enzyme activities in FG cells caused the accumulation of 02 content in FG cells, leading to the change of cell morphology and even the death of cells. The results showed that FG cell line is suitable for the evaluation of the acute toxicity of abamectin.展开更多
Antioxidants play an important role in inhibiting and scavenging free radicals in human, providing protection against cellular damage in relation to cancer initiation. Seaweeds have been proved to have high antioxidan...Antioxidants play an important role in inhibiting and scavenging free radicals in human, providing protection against cellular damage in relation to cancer initiation. Seaweeds have been proved to have high antioxidant activity. Thus, this research was carried out to determine the antioxidant and anticancer properties of edible red seaweed, Gracilaria manilaensis (Gracilariales, Rhodophyta). The extracts were prepared by Soxhlet extraction using organic solvents with different polarities. The antioxidant activities of extracts were determined in terms of their flee radical scavenging activity (RSA IC50) and total phenolic content (TPC). The cytotoxic activity of extracts were tested against human ovarian cancer cell line (Caov-3), human breast cancer cell line (MDA-MB-231 and MCF-7), human cervical cell line (HeLa), mouse fibroblast cell line (L929) and Madin-Darby canine kidney (MDCK) and the cell viability after 72 h incubation was determined by methylene blue assay. The findings showed that acetone extract has the lowest DPPH IC50 value followed by ethyl acetate extract. Both extracts also showed high values of TPC. Dichloromethane extract had the strongest cytotoxic on MDA-MB-231 (53.90 μg/mL ± 5.59 μg/mL) and HeLa (95.50 μg/mL). While, acetone and ethyl acetate extracts were cytotoxic on MCF-7 (66.07 μg/mL) and Caov-3 (69.67 μg/mL ± 13.94 μg/mL). It could be concluded that the antioxidant and cytotoxic activities of G. manilaensis were influenced by the types of solvents used and thus had a potential to develop as a cancer chemoprevention or anticancer agent against selected cancer.展开更多
AIM:To investigate the suppressive activity of MUTYH variant proteins against mutations caused by oxidative lesion,8-hydroxyguanine(8OHG),in human cells.METHODS:p.R154H,p.M255V,p.L360P,and p.P377L MUTYH variants,which...AIM:To investigate the suppressive activity of MUTYH variant proteins against mutations caused by oxidative lesion,8-hydroxyguanine(8OHG),in human cells.METHODS:p.R154H,p.M255V,p.L360P,and p.P377L MUTYH variants,which were previously found in patients with colorectal polyposis and cancer,were selected for use in this study.Human H1299 cancer cell lines inducibly expressing wild-type(WT) MUTYH(type 2) or one of the 4 above-mentioned MUTYH variants were established using the piggyBac transposon vector system,enabling the genomic integration of the transposon sequence for MUTYH expression.MUTYH expression was examined after cumate induction using Western blotting analysis and immunofluorescence analysis.The intracellular localization of MUTYH variants tagged with FLAG was also immunofluorescently examined.Next,the mutation frequency in the supF of the shuttle plasmid pMY189 containing a single 8OHG residue at position 159 of the supF was compared between empty vector cells and cells expressing WT MUTYH or one of the 4 MUTYH variants using a supF forward mutation assay.RESULTS:The successful establishment of human cell lines inducibly expressing WT MUTYH or one of the 4 MUTYH variants was concluded based on the detection of MUTYH expression in these cell lines after treatment with cumate.All of the MUTYH variants and WT MUTYH were localized in the nucleus,and nuclear localization was also observed for FLAG-tagged MUTYH.The mutation frequency of supF was 2.2 × 10-2 in the 8OHG-containing pMY189 plasmid and 2.5 × 10-4 in WT pMY189 in empty vector cells,which was an 86-fold increase with the introduction of 8OHG.The mutation frequency(4.7 × 10-3) of supF in the 8OHG-containing pMY189 plasmid in cells overexpressing WT MUTYH was significantly lower than in the empty vector cells(P < 0.01).However,the mutation frequencies of the supF in the 8OHG-containing pMY189 plasmid in cells overexpressing the p.R154H,p.M255V,p.L360P,or p.P377L MUTYH variant were 1.84 × 10-2,1.55 × 10-2,1.91 × 10-2,and 1.96 × 10-2,respectively,meaning that no significant difference was observed in the mutation frequency between the empty vector cells and cells overexpressing MUTYH mutants.CONCLUSION:The suppressive activities of p.R154H,p.M255V,p.L360P,and p.P377L MUTYH variants against mutations caused by 8OHG are thought to be severely impaired in human cells.展开更多
The Pomegranate (Punica Granatum), which belongs tothe Lythraceae family, has been used for centuries in traditional Greco-Arab and Islamic medicine of its vermifuge properties and also to treat various diseases. Th...The Pomegranate (Punica Granatum), which belongs tothe Lythraceae family, has been used for centuries in traditional Greco-Arab and Islamic medicine of its vermifuge properties and also to treat various diseases. The aim of this research was to investigate the cytotoxicity and antioxidant activities of Moroccan Pomegranate (peel, leaves, branches, flowers and corolla). Further, the biological activities were correlated with phytochemical contents of the plant extracts. Methanolic extract from different parts of Punica Granatum was assessed for its antiproliferative activity in two human cancer (breast and colon) cells lines (MBA-MD 231 and HT-29), through MTT (3-(4,5-dimethyl-2- thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) bioassay using cell viability and cytotoxicity indices. DPPH (2,2-diphenyl-l-picrylhydrazyl) assay was conducted to screen the antioxidant property of the extracts together with its phenolic and flavonoids content were evaluated, as well. The methanolic extract ofPunica Granatum (peel, leaves, branches, flowers and corolla) showed the highest antiproliferative activity on MBA-MD 231 (IC50 was 133.53-233.32 μg/mL) and HT-29 (IC50 was 127.58-203.24 μg/mL) cells. Antioxidants contents are distributed as follows: peel 〉 leaf 〉 flower 〉 corolla 〉 branches. The inhibitory activities required for decreasing initial DPPH by 50% are 8.27, 9.9, 10.06, 11.67 and 13.28 μg/mL, respectively. These results are in correlation with polyphenols content from corolla, peel, leaves, flower and branches are 120.7, 115, 96.65, 90.73 and 64.67 mg GAE/g dw (mg gallic acid equivalents per g dry weight) and flavonoids are 188.8, 221.7, 180.2, 193.7 and 158.5 mg QE/g dw (mg quercetin equivalents per g dry weight). Our results show that the peel, flowers, corolla, leaves and branches of Moroccan Pomegranate may contain a lot of bioactive compounds which are responsible for strong antioxidant and cytotoxicity activities observed here. Our finding indicates the possibility of using the extracts of this plant as source of natural antioxidant and anticancer mainly for its abundant phenolic and flavonoid contents.展开更多
OBJECTIVE: To determine the cardioprotective ef- fect of magnesium lithospermate B (MLB) on myo- cardial ischemia/reperfusion (MI/R) injury and to in- vestigate the antioxidant potential in vivo and in vitro. MET...OBJECTIVE: To determine the cardioprotective ef- fect of magnesium lithospermate B (MLB) on myo- cardial ischemia/reperfusion (MI/R) injury and to in- vestigate the antioxidant potential in vivo and in vitro. METHODS: MI/R injury was induced by the occlu- sion of left anterior descending coronary artery for 30 min followed by reperfusion for 3 h in rats. After reperfusion, hearts were harvested to assess infarct size, histopathological damages, the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH) and malondialdehyde (MDA). Blood samples were collected to determine serum levels of creatine kinase-MB (CK-MB), cardiac troponin (cTnl) and lactate dehydrogenase (LDH). Furthermore, simulatedischemia/reperfusion (SI/R) injury in vitro was established by oxygen and glucose deprivation (OGD) for 2 h followed by 24-hour recovery period in cardiomyocytes. The activity of LDH in the cultured su- pernatant and the levels of intracellular reactive oxygen species (ROS), SOD and MDA in cardiomyo- cytes were also measured. Finally, cardiomyocytes apoptosis was determined with flow cytometry. RESULTS: MLB significantly limited infarct size, ameliorated histopathological damages and prevented leakage of CK-MB, cTnl and LDH. Additional- ly, SOD, CAT, GPx and GSH activities were notably increased by MLB, along with the MDA content decreased as compared with the model group in rats. In vitro study, MLB also decreased LDH activity in the cultured supernatant, increased SOD activity in cardiomyocytes, reduced intracellular ROS and MDA levels, and significantly suppressed cardiomyocytes apoptosis. CONCLUSION: MLB possessed remarkably cardioprotective effects on MI/R injury in vivo and in vitro. The protection of MLB may contribute to its antioxidant properties.展开更多
The polysaccharides from pumpkin fruit (PP) were obtained and purified by hot-water extraction, anion-exchange chromatography, and gel column chromatography. The physicochemical properties of PP were determined by g...The polysaccharides from pumpkin fruit (PP) were obtained and purified by hot-water extraction, anion-exchange chromatography, and gel column chromatography. The physicochemical properties of PP were determined by gel filtration chromatography, gas chromatography, fourier transform infrared (FTIR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. Results indicated that the molecular weight of PP was about 23 kDa and PP was composed of D-Arabinose, D-Marmose, D-Glucose, and D-Galactose with a molar ratio of 1 : 7.79 : 70.32 : 7.05. FTIR and NMR spectra indicated that PP was the polysaccharide containing pyranose ring. Additionally, PP protected islets cells from streptozotocin (STZ) injury in vitro via increasing the levels of super-oxide dismutase (SOD) and malondialdehyde (MDA) and reducing the production of NO. The experiment of reverse transcriptase-polymerase chain reaction further proved that PP inhibited apoptosis via modulating the expression of Bax/Bcl-2 in STZ-damaged islet cells. In conclusion, PP could be explored as a novel agent for the treatment of diabetes mellitus.展开更多
Controlled ecological life support systems provide food, air, water, and other basic living resources for crew members on long-duration spaceflight missions. Plants are an important basic requirement of these systems ...Controlled ecological life support systems provide food, air, water, and other basic living resources for crew members on long-duration spaceflight missions. Plants are an important basic requirement of these systems and their biological characteristics in space have very high research value. Based on experiments of spaceflight in Shenzhou 8 spacecraft and simulating microgravity effects on three-dimensional (3-D) clinostat, the biological characteristics of tomato's leaf cell sub-microstructure and antioxidant enzyme activities were studied and compared in this work. Results showed that leaf cell sub-microstructure of the tomato samples experiencing spaceflight had more changes than effects, and both peroxidase (POD) and superoxide dismutase (SOD) that of the samples processed by simulated microgravity activities increase obviously in both the environments.展开更多
Xiangxi flavor vinegar(XV) is one of Hunan Province's traditional fermented vinegars.It is produced from herb,rice,and spring water with spontaneous liquid-state fermentation techniques.In this study,we investigate...Xiangxi flavor vinegar(XV) is one of Hunan Province's traditional fermented vinegars.It is produced from herb,rice,and spring water with spontaneous liquid-state fermentation techniques.In this study,we investigated the antioxidant property of XV by analyzing its antioxidant compounds,its free radical scavenging property in vitro and in vivo,and its effects on antioxidant enzyme activity and apoptosis in Caenorhabditis elegans.The results showed that XV is rich in antioxidants.In particular,ligustrazine reached 6.431 μg/ml.The in vitro 2,2-diphenyl-1-picrylhydrazyl free radical(DPPH*),hydroxyl radical(*OH),and superoxide anion radical(O2^(*-)) scavenging rates of XV were 95.85%,97.22%,and 63.33%,respectively.The reactive oxygen species(ROS) content in XV-treated C.elegans decreased significantly(P0.01) compared to the control group.Giutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and catalase(CAT) activities were remarkably increased(P0.01) in C.elegans after XV treatment.In addition,XV could upreguiate CED-9 protein expression and downreguiate CED-3 protein expression in C.elegans.These results prove that XV is rich in antioxidants and scavenges radicals in vitro efficiently.XV inhibits apoptosis in C.elegans probably by scavenging ROS and increasing the activities of its antioxidant enzymes.展开更多
文摘An acidic polysaccharide, H2, was isolated from the alkali-extract CHC of seeds of Cuscuta chinensis Lam. with the molecular weight more than 1.0 x 10(6). Chemical and spectroscopic studies led to the structure determination as follows: the backbone chain consists of 1, 6-linked-beta -D Galp, 1,4-linked-X -D Galp, 1,4-linked-beta -D GalA and 1,2- or 1,4-linked-beta -L Rhap having branching points at position O-3 of some 1,6-linked-beta -D Galp residues (one among eight) and O-4 or O-2 of 1,2- or 1,4-linked-beta -L Rhap residues to terminal beta -D-galactopyranose. The side chains composed of terminal Galp, 1,6-linked-beta -D Galp, 1,4-linked beta -D Galp and 1,3,6-linked-beta -D Galp also linked at position O-3 of 1,6-linked-beta -D Galp residues in the backbone chain. beta -L-arabinofuranosyl and terminal beta -L-rhamnopyranosyl residues existed in the periphery of this polysaccharide linked to O-3 of 1,6-linked-beta -D Galp residues in the backbone chain and the side chains. The polysaccharide H2 increased significantly the survival rate of PC12 cells indicating that it had protective effects against H2O2 insult.
文摘Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW264.7) were stimulated by LPS alone, or with long-term of PMA pretreatment. Then cytotoxicities to P815 cells (by MTT assay) and IL-1, TNF- (by ELISA) and nitric oxide (NO) production (by Griess reagent) in supernatants were measured. Western blot for PKC isoforms after long-term PMA pretreatment was analyzed. Results: RAW264.7 cells were stimulated with LPS to kill target tumor cells P815, whereas P388D1 cells failed to develop such an ability. Down-regulation of PKC isoforms by chronic treatment with PMA significantly inhibited the LPS-induced cytotoxicity in RAW264.7 cells. In unstimulated state, Western blotting with rabbit antiserum specific for the PKC, 1, 2, or showed all 5 isoforms were detected in P388D1 cells, while only PKC, PKC1 and PKC were detected in RAW264.7 cells. Exposure of the cells to long-term of PMA treatment significantly down-regulated the expression of PKC, PKC1 and PKC in RAW264.7 cells. But in P388D1 cells, although PKC, PKC and PKC were down-regulated, the expression of PKC1 and PKC2 could not be regulated. Comparing with LPS-induced IL-1, TNF- and NO production by the two macrophage cell lines, P388D1 failed to produce NO. In RAW264.7 cells, LPS-induced NO production and antitumor activity was attenuated by the addition of L-NAME, an iNOS inhibitor. Conclusion: The results indicated a critical role of PKC in LPS-induced antitumor activity and this cytotoxicity is mainly due to PKC- mediated NO production by RAW264.7 cells, but not a direct cytotoxic activity.
基金Supported by The National Scientific Research Fund of China(2008JK007)the National Key Research and Development Program of China for the Tenth Five-Year Plan,No.2006BAD06A14
文摘AIM:To investigate the antioxidant activity of chitooligosaccharides(COSs)on pancreatic islet cells in diabetic rats induced by streptozotocin. METHODS:The antioxidant effect of COSs on pancreatic islet cells was detected under optical microscopy and with colorimetric assay and gel electrophoresis.The activities of glutathione peroxidase and superoxide dismutase,total antioxidant capacity,and content of malondialdehyde in serum and tissue slices of pancreas were examined after 60 d to determine the effect of COSs in streptozotocin-induced diabetes in rats. RESULTS:COSs can prohibit the apoptosis of pancreatic islet cells.All concentrations of COSs can improve the capability of total antioxidant capacity and activity of superoxide dismutase and decrease the content of malondialdehyde drastically.Morphological investigation in the pancreas showed that COSs have resulted in the reduction of islets,loss of pancreatic cells,and nuclear pyknosis of pancreatic cells. CONCLUSION:COSs possess various biological activities and can be used in the treatment of diabetes mellitus.
文摘Drug-induced liver injury is a significant and still unresolved clinical problem. Limitations to knowledge about the mechanisms of toxicity render incomplete the detection of hepatotoxic potential during preclinical development. Several xenobiotics are lipophilic substances and their transformation into hydrophilic compounds by the cytochrome P-450 system results in production of toxic metabolites. Aging, preexisting liver disease, enzyme induction or inhibition, genetic variances, local 02 supply and, above all, the intrinsic molecular properties of the drug may affect this process. Necrotic death follows antioxidant consumption and oxidation of intracellular proteins, which determine increased permeability of mitochondrial membranes, loss of potential, decreased ATP synthesis, inhibition of Ca^2+-dependent ATPase, reduced capability to sequester Ca^2+ within mitochondria, and membrane bleb formation. Conversely, activation of nucleases and energetic participation of mitochondria are the main intracellular mechanisms that lead to apoptosis. Non-parenchymal hepatic cells are inducers of hepatocellular injury and targets for damage. Activation of the immune system promotes idiosyncratic reactions that result in hepatic necrosis or cholestasis, in which different HLA genotypes might play a major role. This review focuses on current knowledge of the mechanisms of drug-induced liver injury and recent advances on newly discovered mechanisms of liver damage. Future perspectives including new frontiers for research are discussed.
文摘The cytotoxicity of abamectin to the Gill Cell Line of Flounder (FG cell line) was examined in this study. It was found that the exposure of FG cells to abamectin caused the decreases of both cell growth rate and antioxidant enzyme activities, and the increase of intracellular 02 content. It was proposed that the reduction of antioxidant enzyme activities in FG cells caused the accumulation of 02 content in FG cells, leading to the change of cell morphology and even the death of cells. The results showed that FG cell line is suitable for the evaluation of the acute toxicity of abamectin.
文摘Antioxidants play an important role in inhibiting and scavenging free radicals in human, providing protection against cellular damage in relation to cancer initiation. Seaweeds have been proved to have high antioxidant activity. Thus, this research was carried out to determine the antioxidant and anticancer properties of edible red seaweed, Gracilaria manilaensis (Gracilariales, Rhodophyta). The extracts were prepared by Soxhlet extraction using organic solvents with different polarities. The antioxidant activities of extracts were determined in terms of their flee radical scavenging activity (RSA IC50) and total phenolic content (TPC). The cytotoxic activity of extracts were tested against human ovarian cancer cell line (Caov-3), human breast cancer cell line (MDA-MB-231 and MCF-7), human cervical cell line (HeLa), mouse fibroblast cell line (L929) and Madin-Darby canine kidney (MDCK) and the cell viability after 72 h incubation was determined by methylene blue assay. The findings showed that acetone extract has the lowest DPPH IC50 value followed by ethyl acetate extract. Both extracts also showed high values of TPC. Dichloromethane extract had the strongest cytotoxic on MDA-MB-231 (53.90 μg/mL ± 5.59 μg/mL) and HeLa (95.50 μg/mL). While, acetone and ethyl acetate extracts were cytotoxic on MCF-7 (66.07 μg/mL) and Caov-3 (69.67 μg/mL ± 13.94 μg/mL). It could be concluded that the antioxidant and cytotoxic activities of G. manilaensis were influenced by the types of solvents used and thus had a potential to develop as a cancer chemoprevention or anticancer agent against selected cancer.
基金Supported by Grants from the Ministry of Health,Labour and Welfare(21-1)the Japan Society for the Promotion of Science (22590356 and 22790378)+3 种基金the Hamamatsu Foundation for Science and Technology Promotion,the Ministry of Education, Culture,Sports,Science and Technology(221S0001)the Takeda Science Foundationthe Aichi Cancer Research Foundationthe Smoking Research Foundation
文摘AIM:To investigate the suppressive activity of MUTYH variant proteins against mutations caused by oxidative lesion,8-hydroxyguanine(8OHG),in human cells.METHODS:p.R154H,p.M255V,p.L360P,and p.P377L MUTYH variants,which were previously found in patients with colorectal polyposis and cancer,were selected for use in this study.Human H1299 cancer cell lines inducibly expressing wild-type(WT) MUTYH(type 2) or one of the 4 above-mentioned MUTYH variants were established using the piggyBac transposon vector system,enabling the genomic integration of the transposon sequence for MUTYH expression.MUTYH expression was examined after cumate induction using Western blotting analysis and immunofluorescence analysis.The intracellular localization of MUTYH variants tagged with FLAG was also immunofluorescently examined.Next,the mutation frequency in the supF of the shuttle plasmid pMY189 containing a single 8OHG residue at position 159 of the supF was compared between empty vector cells and cells expressing WT MUTYH or one of the 4 MUTYH variants using a supF forward mutation assay.RESULTS:The successful establishment of human cell lines inducibly expressing WT MUTYH or one of the 4 MUTYH variants was concluded based on the detection of MUTYH expression in these cell lines after treatment with cumate.All of the MUTYH variants and WT MUTYH were localized in the nucleus,and nuclear localization was also observed for FLAG-tagged MUTYH.The mutation frequency of supF was 2.2 × 10-2 in the 8OHG-containing pMY189 plasmid and 2.5 × 10-4 in WT pMY189 in empty vector cells,which was an 86-fold increase with the introduction of 8OHG.The mutation frequency(4.7 × 10-3) of supF in the 8OHG-containing pMY189 plasmid in cells overexpressing WT MUTYH was significantly lower than in the empty vector cells(P < 0.01).However,the mutation frequencies of the supF in the 8OHG-containing pMY189 plasmid in cells overexpressing the p.R154H,p.M255V,p.L360P,or p.P377L MUTYH variant were 1.84 × 10-2,1.55 × 10-2,1.91 × 10-2,and 1.96 × 10-2,respectively,meaning that no significant difference was observed in the mutation frequency between the empty vector cells and cells overexpressing MUTYH mutants.CONCLUSION:The suppressive activities of p.R154H,p.M255V,p.L360P,and p.P377L MUTYH variants against mutations caused by 8OHG are thought to be severely impaired in human cells.
文摘The Pomegranate (Punica Granatum), which belongs tothe Lythraceae family, has been used for centuries in traditional Greco-Arab and Islamic medicine of its vermifuge properties and also to treat various diseases. The aim of this research was to investigate the cytotoxicity and antioxidant activities of Moroccan Pomegranate (peel, leaves, branches, flowers and corolla). Further, the biological activities were correlated with phytochemical contents of the plant extracts. Methanolic extract from different parts of Punica Granatum was assessed for its antiproliferative activity in two human cancer (breast and colon) cells lines (MBA-MD 231 and HT-29), through MTT (3-(4,5-dimethyl-2- thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) bioassay using cell viability and cytotoxicity indices. DPPH (2,2-diphenyl-l-picrylhydrazyl) assay was conducted to screen the antioxidant property of the extracts together with its phenolic and flavonoids content were evaluated, as well. The methanolic extract ofPunica Granatum (peel, leaves, branches, flowers and corolla) showed the highest antiproliferative activity on MBA-MD 231 (IC50 was 133.53-233.32 μg/mL) and HT-29 (IC50 was 127.58-203.24 μg/mL) cells. Antioxidants contents are distributed as follows: peel 〉 leaf 〉 flower 〉 corolla 〉 branches. The inhibitory activities required for decreasing initial DPPH by 50% are 8.27, 9.9, 10.06, 11.67 and 13.28 μg/mL, respectively. These results are in correlation with polyphenols content from corolla, peel, leaves, flower and branches are 120.7, 115, 96.65, 90.73 and 64.67 mg GAE/g dw (mg gallic acid equivalents per g dry weight) and flavonoids are 188.8, 221.7, 180.2, 193.7 and 158.5 mg QE/g dw (mg quercetin equivalents per g dry weight). Our results show that the peel, flowers, corolla, leaves and branches of Moroccan Pomegranate may contain a lot of bioactive compounds which are responsible for strong antioxidant and cytotoxicity activities observed here. Our finding indicates the possibility of using the extracts of this plant as source of natural antioxidant and anticancer mainly for its abundant phenolic and flavonoid contents.
基金Supported by the National Natural Science Foundation of China (No. 81173514,No.81001673)the Xijing Research Boosting Program (No. XJZT10D02)the Excellent Civil Service Training Fund of Fourth Military Medical University(No. 4138C4IDK6)
文摘OBJECTIVE: To determine the cardioprotective ef- fect of magnesium lithospermate B (MLB) on myo- cardial ischemia/reperfusion (MI/R) injury and to in- vestigate the antioxidant potential in vivo and in vitro. METHODS: MI/R injury was induced by the occlu- sion of left anterior descending coronary artery for 30 min followed by reperfusion for 3 h in rats. After reperfusion, hearts were harvested to assess infarct size, histopathological damages, the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH) and malondialdehyde (MDA). Blood samples were collected to determine serum levels of creatine kinase-MB (CK-MB), cardiac troponin (cTnl) and lactate dehydrogenase (LDH). Furthermore, simulatedischemia/reperfusion (SI/R) injury in vitro was established by oxygen and glucose deprivation (OGD) for 2 h followed by 24-hour recovery period in cardiomyocytes. The activity of LDH in the cultured su- pernatant and the levels of intracellular reactive oxygen species (ROS), SOD and MDA in cardiomyo- cytes were also measured. Finally, cardiomyocytes apoptosis was determined with flow cytometry. RESULTS: MLB significantly limited infarct size, ameliorated histopathological damages and prevented leakage of CK-MB, cTnl and LDH. Additional- ly, SOD, CAT, GPx and GSH activities were notably increased by MLB, along with the MDA content decreased as compared with the model group in rats. In vitro study, MLB also decreased LDH activity in the cultured supernatant, increased SOD activity in cardiomyocytes, reduced intracellular ROS and MDA levels, and significantly suppressed cardiomyocytes apoptosis. CONCLUSION: MLB possessed remarkably cardioprotective effects on MI/R injury in vivo and in vitro. The protection of MLB may contribute to its antioxidant properties.
基金supported by the Natural Science Foundation Committee of China(NSFC 81202467)the Natural Science Foundation of Jiangsu Province in China(BK2012232)+2 种基金the Natural Science Research Project of Universities in Jiangsu Province of China(11KJB350004,06KJD310150,12KJB310012)the Natural Science Foundation of Nantong City of China(BK2011044)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘The polysaccharides from pumpkin fruit (PP) were obtained and purified by hot-water extraction, anion-exchange chromatography, and gel column chromatography. The physicochemical properties of PP were determined by gel filtration chromatography, gas chromatography, fourier transform infrared (FTIR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. Results indicated that the molecular weight of PP was about 23 kDa and PP was composed of D-Arabinose, D-Marmose, D-Glucose, and D-Galactose with a molar ratio of 1 : 7.79 : 70.32 : 7.05. FTIR and NMR spectra indicated that PP was the polysaccharide containing pyranose ring. Additionally, PP protected islets cells from streptozotocin (STZ) injury in vitro via increasing the levels of super-oxide dismutase (SOD) and malondialdehyde (MDA) and reducing the production of NO. The experiment of reverse transcriptase-polymerase chain reaction further proved that PP inhibited apoptosis via modulating the expression of Bax/Bcl-2 in STZ-damaged islet cells. In conclusion, PP could be explored as a novel agent for the treatment of diabetes mellitus.
基金supported by the National Basic Research Program of China("973"Project)(Grant No.2011CB710902)the National Manned Spaceflight Project of China
文摘Controlled ecological life support systems provide food, air, water, and other basic living resources for crew members on long-duration spaceflight missions. Plants are an important basic requirement of these systems and their biological characteristics in space have very high research value. Based on experiments of spaceflight in Shenzhou 8 spacecraft and simulating microgravity effects on three-dimensional (3-D) clinostat, the biological characteristics of tomato's leaf cell sub-microstructure and antioxidant enzyme activities were studied and compared in this work. Results showed that leaf cell sub-microstructure of the tomato samples experiencing spaceflight had more changes than effects, and both peroxidase (POD) and superoxide dismutase (SOD) that of the samples processed by simulated microgravity activities increase obviously in both the environments.
基金supported by the College of Food Science and Technology,Hunan Agricultural University,China
文摘Xiangxi flavor vinegar(XV) is one of Hunan Province's traditional fermented vinegars.It is produced from herb,rice,and spring water with spontaneous liquid-state fermentation techniques.In this study,we investigated the antioxidant property of XV by analyzing its antioxidant compounds,its free radical scavenging property in vitro and in vivo,and its effects on antioxidant enzyme activity and apoptosis in Caenorhabditis elegans.The results showed that XV is rich in antioxidants.In particular,ligustrazine reached 6.431 μg/ml.The in vitro 2,2-diphenyl-1-picrylhydrazyl free radical(DPPH*),hydroxyl radical(*OH),and superoxide anion radical(O2^(*-)) scavenging rates of XV were 95.85%,97.22%,and 63.33%,respectively.The reactive oxygen species(ROS) content in XV-treated C.elegans decreased significantly(P0.01) compared to the control group.Giutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and catalase(CAT) activities were remarkably increased(P0.01) in C.elegans after XV treatment.In addition,XV could upreguiate CED-9 protein expression and downreguiate CED-3 protein expression in C.elegans.These results prove that XV is rich in antioxidants and scavenges radicals in vitro efficiently.XV inhibits apoptosis in C.elegans probably by scavenging ROS and increasing the activities of its antioxidant enzymes.