侧柏叶多糖的提取及其抗氧化活性的研究

目的:提取侧柏叶多糖,并分析其抗氧化活性。方法:以蒸馏水为提取剂,采用超声波法提取侧柏叶多糖,通过单因素及正交实验研究对多糖提取率影响的主要因素,并确定其最佳提取工艺条件。同时通过对侧柏叶多糖清除超氧阴离子自由基的测定考察多糖的抗氧化能力。结果:超声提取法的优化工艺条件为:料液比(g:m L)为1:35,提取温度为60℃,超声功率为180 W,作用时间为40 min。在此条件下,侧柏叶多糖的最佳提取率为8.35%。侧柏叶多糖对超氧阴离子有一定的清除作用,且其作用强度随侧柏叶多糖浓度增大而增加。结论:超声波强化提取侧柏叶多糖效果省时高效,侧柏叶多糖具有较强的抗氧化活性。

Antioxidant phenanthrenes and lignans from Dendrobium nobile

To study the antioxidant constituents from the stems of Dendrobium nobile, and to discuss theft structure-activity relationship. Compounds were isolated from a 60% ethanolic extract by various chromatographic techniques and were identified by spectral analysis. The antioxidant activities of compounds were evaluated by DPPH free radical scavenging assay. Five phenanthrenes and four lignans were obtained from the active fractions ofD. nobile. Their structures were identified as fimbriatone （1）, confusarin （2）, flavanthrinin （3）, 2,5-dihydroxy-4,9-dimethoxyphenanthrene （4）, 3,7-dihydroxy-2,4-dimethoxyphenanthrene （5）, syringaresinol （6）, pinoresinol （7）, medioresinol （8） and lirioresinol-A （9）, respectively. Compounds 2 and 6 exhibited more potent DPPH scavenging activities than vitamin C. All the above compounds were reported from this plant for the first time, and compounds 3, 4 and 9 were reported for the first time from the genus of Dendrobiurn. For all phenanthrenes and lignans, an electron-donating methoxyl group in the ortho position to the phenolic hydroxyl group exhibits enhanced antioxidant activities.

Antioxidant Activity of Polysaccharides in Yam Bulbils and Their Hypoglycemic Effect in Diabetic Mice

The polysaccharides in yam bulbils were extracted, and their antioxidant activity and hypoglycemic effect in diabetic mice were discussed. The results showed that the antioxidant activity of polysaccharides in yam bulbils was signifi- cantly enhanced with the increase of concentration; they showed a strong scaveng- ing ability against DPPH. and .OH, and the scavenging ability was dose dependent to some extent; the scavenging rates reached 91.15% and 89.06% respectively when the dose reached 4.0 mg/ml; the polysaccharides in yam bulbils significantly educed the blood glucose in model rice, and the hypoglycemic effect of large-dose polysaccharides was more obvious. The polysaccharides in yam bulbils has good antioxidant activity and hypoglycemic effect, which provides a new source for devel- opment of safe and natural food antioxidants and blood sugar-lowering agents.

Factors Influencing the Effects of Theabrownin in Pu-erh Tea on Scavenging DPPH Radicals

[Objective] This study aimed to explore factors influencing the effects of theabrownin in Pu-erh tea on scavenging DPPH radicals. [Method] Taking Pu-erh tea as main research object, theabrownin in Pu-erh tea from tea piles with different times of mixture, stored for different years and processed by both the old and new techniques were extracted and isolated. And DPPH method was used to study the effects of theabrownin solution and tea liquor on scavenging radicals and their antioxidant activity. [Result] The result showed that the theabrownin content in Pu-erh tea from tea piles with one to four times of mixture revealed a first declining and subsequently rising trend. Under different pH conditions, the rates of DPPH radical scavenging of 1 mg/ml theabrownin solution made from tea piles with different times of mixture decreased gradually with the increase of pH, and all the rates were lower than that of the control vitamin C solution; the rates of DPPH radical scavenging of theabrownin from tea samples stored for different years and produced by different techniques dropped slowly within pH range 3.0-6.0; when pH was higher than 6.0, the rates of DPPH radical scavenging of theabrownin from tea samples produced by both new and traditional techniques in 2001 declined obviously, while the declining trend was not obvious in either the black tea or the Pu-erh tea produced by new technique in 2005. Under different theabrownin concentrations, the rates of DPPH radical scavenging of theabrownin solution made from tea piles with different times of mixture ascended as the concentration increased and the highest rate occurred in tea pile with four times of mixture, reaching up to 46.56%±0.88%; for tea samples stored for different years and produced by different techniques, the rates of DPPH radical scavenging of theabrownin rose gradually with the increase of the theabrownin concentration; the rate of radical scavenging of theabrownin in the black tea ascended obviously and that in Pu-erh tea was decreasing within the concentration range of 0.04 -0.06 mg/ml. At pH6.0, the DPPH radical scavenging ability of theabrownin in 0.1 mg/ml theabrownin solution and tea liquor containing 0.1mg/ml theabrownin from tea piles with different times of mixture ascended with the time of mixture; solution made from tea samples produced by different techniques in different years, the DPPH radical scavenging ability of theabrownin solution and tea liquor followed： Pu-erh tea produced by the new technique in 2001 ﹥ Pu-erh tea produced by the old technique in 2001﹥ Pu-erh tea produced by the new technique in 2005. And the DPPH radical scavenging ability of theabrownin in tea liquor containing 0.1 mg/ml theabrownin was higher than that in 0.1 mg/ml theabrownin Solution made from tea samples either with different times of mixture or produced by different techniques in different years. The above results indicated that many factor influenced the effects of theabrownin on eliminating DPPH radicals, including the time of fermentation, fermentation technique, years of storage, concentration and pH all had great influence on the effects of theabrownin on eliminating radicals. To ensure a good effect on scavenging radicals, it is better to prepare theabrownin at pH6.0 with concentration between 0.08 and 0.1 mg/ml for drinking. [Conclusion] Results of this study provide a basis for developing functional products of theabrownin from Pu-erh tea in the future.

Research on Oxidative Stress Induced by Tenuazonic Acid from Alternaria augustiovoide and Changes in Antioxidant Enzyme Activities in Leaves of Echinochloa crus-galli

[Objective] The aim of the study was to determine whether phytotoxicity of TeA against Echinochloa crus-galli leaves was correlated with oxidative stress caused by generation of reactive oxygen and the changes of antioxidant enzymes activity.[Method] The changes of malondialdehyde（MDA）content,hydrogen peroxide（H2O2）,and activities of superoxide dismutase（SOD）,glutathione reductase（GR）and catalase（CAT）were studied by leaf segment method in vitro.[Result] After the treatment of 500 μmol/L TeA,the content of MDA and H2O2 increased by 247.86% and 67.00%,respectively,indicating that the accumulation of MDA and H2O2 in E.crus-galli leaves was due to the reactive oxygen burst induced by TeA.TeA induced a significant increase in activities of SOD,GR and CAT.At 500 μmol/L TeA,activities of SOD,GR and CAT increased more than one fold compared with the control.[Conclusion] TeA could not only cause oxidative stress in leaves of E.crus-galli through the induction of reactive oxygen,but also induce the increasing of antioxidant enzyme activity.

Study on Anti-oxidation and Inhibitory Effect on Nonenzymatic Glycation Reaction of Fermentation Extract from Biotransformation of Ginkgo biloba L. (EGB) by Hericium erinaceus

[ Objective] In order to study the anti-oxidation and inhibitory effect on nonenzymatic glycation reaction of EGB fermentation extraction biotransformed by Hericium erinaceus. [ Method ] The free radical scavenging ability and reducing capacity of DPPH as well as inhibitory rate of nonenzymatic glycation reaction were measured targets for comparing changes of anti-oxidation and inhibitory effect on nonenzymatic glycation reaction of fermentation lyophilizer and fermentation extraction before and after EGB fermention adsorbed by AB-8 macroporous resin. The EGB fermention was biotransformed by Hericium erinaceus. [ Result ] After adsorbed by AB-8 macroporous resin, the bioactive matters were concentrated and separated. The free radical scavenging rate, reducing capacity and inhibitory rate of nonenzymatic glycation reaction were increased significantly after adsorbed by AB-8 macroporous resin. [ Conclusion] AB-8 macroporous resin could be used for preliminary concentration of EGB fermentation which was biotransformed by Hericium erinaceus.

An Antioxidant Acidic Polysaccharide from Cuscuta chinensis

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.

Novel Stilbene Glycosides from Polygonum multiflorum

Two new stilbene glycosides (1 and 2), together with nine known compounds (3-11), were isolated from the water extract of Polygonum multiflorum Thunb. The structures of the new compounds were elucidated by their chemical properties and spectroscopic analyses, including extensive 2D NMR experiments. Compound 2 showed strong DNA cleavage activity, and compounds 1, 2 and 10 (2, 3, 4′, 5_tetrahydroxy_ trans _stilbene_2_O_β_ D _glucopyranoside) exhibited significant inhibition of lipid peroxidation.

Antioxidase Activities of Tetraploid Lines and Diploid Control Line of Chrysanthemum cinerariifolium

Objective The SOD, POD and APX enzyme activities of tetraploid lines and diploid control line of Chrysanthemum cinerariifolium in test-tube and field were determined in order to provide a reference for the selective breeding of high resistance good tetraploid varieties of C. cinerariifolium. Method The SOD, POD and APX activities of each sample were determined. Every material should have 3 replicates which been determined and calculated mean value to analyze. Result The various kinds of antioxidase activities in protective enzyme system of tetraploid lines were increased generally in comparison with that of diploid control line. And there is certain relativity between the samples in field and in test-tube of these lines. Conclusion The antioxidase activities of each line in test-tube could be considered as reference index to screen high resistance good tetraploid varieties of C. cinerariifolium.

Effects of Water Deficit and Increased Nitrogen Application in the Late Growth Stage on Physiological Characters of Anti-aging of Leaves in Different Hybrid Rice Varieties

[Objective] This study aimed to explore the effects of water and fertilizer regulation on the function of leaves in the filling stage. [Method] Six hybrid rice vari- eties including prematurely aged types and non-prematurely aged ones were selected as experimental materials. Limiting water treatment （with soil water potential of about -25 kpa） and increasing nitrogen treatment （additional 10% of granular nitrogen fertiliz- er in addition to limiting water treatment） were applied after heading to analyse the rates of aging and physiological responses of anti-aging in different hybrid rice vari- eties under water deficit and increased nitrogen conditions taking regular water and fertilizer as control. [Result] The results showed that water deficit accelerated the leaf senescence, and prematurely aged types-＇Zhongyou838＇, ＇Tianyou998＇ and ＇Long- ping601 ＇-were more markedly affected by water deficit, of which the rates of chloro- phyll degradation were 6%-8% higher compared to that in another three hybrids. In- creasing nitrogen treatment raised chlorophyll content and slowed down its degrada- tion. Water deficit caused the increase of abscisic acid （ABA） content to obviously varying degrees in leaves of six hybried rice varieties. Responses of ABA content in six hybried rice varieties to increased nitrogen fertilizer were not consistent. Except in ＇Zhongyou838＇, ABA content in the other five hybrids had varying degrees of in- crease. The responses of the activity of antioxidant enzymes in different hybried rice varieties were inconsistent. In response to increased nitrogen fertilizer in combination with water deficit, the activity of each antioxidant enzyme changed differently. Water deficit enhanced the accumulation of malondialdehyde （MDA） in leaves of each hybrid rice. The correlation analysis showed that chlorophyll content was extremely signifi- cantly positively correlated to the resistance of each antioxidant enzyme; SOD activity in leaves also positively related to ABA content. [Conclusion] The rate of leaf aging, physiological activity of anti-aging, and response to water deficit varied in different hy- bried rice varieties. The activity of antioxidant enzymes were not all induced to improve by ABA. SOD activity can be an indicator of resistance to stress. Increasing nitrogen ap- plication significantly postponed leave senescence in the late growth stage of rice.

Differential Expression of PKC Isoforms and Their Tumoricidal Activity in Two Macrophage Cell Lines: Involvement of Nitric Oxide-dependent Mechanisms

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.

Advances in Melatonin and Its Functions in Plants

Abstract Melatonin （N-acetyi-5-methoxytryptamine） is a well-known animal hormone, which is synthesized and secreted by pineal gland and takes part in the regulation of circadian rhythm in animals. At present it has been observed that melatonin is widely existed in higher plants while there have no enough studies on functions of melatonin in plants. Researches have already indicated that the possible functions of melatonin in plants include regulating photoperiod, participating in growth regulation, clearing active oxygen, and promoting activity of antioxidase. General reviews upon functions of melatonin in plants are made upon experiments in recent years. We fo-cus on the demonstrated and predicted biological functions of melatonin in plants to bring researchers up to date on this field. The weakness in present studies and the main research directions are also pointed out.

Effects of Heavy-ion Beams Irradiation on Survival Rate and Antioxidant Enzymes of Sweet Sorghum Seedlings

[Objective] This study aimed to investigate the effects of heavy-ion beams irradiation on the seed germination potential, survival rate, antioxidant enzyme activi- ties and lipid peroxidation of sweet sorghum. [Method] The dry seeds were irradiated by ＇2（36. heavy ion beams with absorbed doses： 0, 40, 80, 120, 160 and 200 Gy, respectively. Then, the seed germination potential, survival rate, antioxidant enzyme activities and lipid peroxidation of sweet sorghum were measured. [Result] Heavy-ion beams irradiation exhibited different influence on germination potential and survival rates. Germination rate showed a downward trend, but the corresponding survival curve of seedlings was saddle-shaped. The activities of SOD, POD, CAT and ASA- POD changed in different trends as well. The MDA content rose toward increasing irradiation dose, suggesting that high dose of heavy-ion beams irradiation enhanced the damage to membrane of sweet sorghum seedlings. [Conclusion] After being irra- diated, germination potential and survival rates of sweet sorghum were decreased, and antioxidant enzymes activity changed greatly. This study laid the basis for fur- ther work on breeding and improvement of sweet sorghum irradiated by ,^（12）C^（6＋） heavy ion beams.

Water Soluble Active Components of Salvia miltiorrhiza and Related Plants

Chemical studies on nine Salvia species yielded various polyphenolic acids. Eleven of them were depsides of R (+) β (3,4 dihydroxyphenyl) lactic acid and a caffeic acid derivative or caffeic acid dimer. Except the two known depsides, rosmarinic acid and lithospermic acid, this type of depsides has not been isolated from other plant materials before, so they were named salvianolic acid A, B, C, D, E, H, I, J and isosalvianolic acid C. Salvianolic acid F, G and przewalskinic acid A were new polyphenolic acids. Pharmacological studies of these polyphenolic acids showed potent antioxidant activities. The effects of these components in protecting against brain and heart damage have been studied.

Effect of Lonicerae Flos extracts on reflux esophagitis with antioxidant activity

AIM： To observe the effects of traditional antiinflammatory medicine Lonicerae FIos （LF） on rat reflux esophagitis （RE） induced by pylorus and forestomach ligation compared with the well-known proton antioxidant, α-tocopherol. METHODS： Rats were pretreated with three different dosages of LF （500, 250 and 125 mg/kg） orally, once a day for 14 d before pylorus and forestomach ligation. Nine hours after pylorus and forestomach ligation, changes to the stomach and esophagus lesion areas, gastric volumes, acid and pepsin outputs, antioxidant effects, esophageal lipid peroxidation, superoxide dismutase （SOD）, catalase （CAT）, malondialdehyde （MDA）, myeloperoxidase and glutathione （GSH） levels, and collagen contents （marker of flexibility） were observed on the esophageal and fundic histopathology. The results were compared with an α-tocopherol （once orally, 1 h before operation, 30 mg/kg） treated group in which the effects on RE were already confirmed.RESULTS： Pylorus and forestomach ligations caused marked increases of gross esophageal and gastric mucosa lesion areas, which corresponded with histopathological changes. In addition, increases of esophageal lipid peroxidation, decreases of SOD, CAT, and GSH-free radical scavengers, increases of collagen were observed. However, these pylorus and forestomach ligation induced RE were dose-dependently inhibited by treatment of 500, 250 and 125 mg/kg of LF extract, mediated by antioxidant effects. RE at 250 mg/kg showed similar effects α-tocopherol. CONCLUSION： The results suggest that antioxidant effects of LF could attenuate the severity of RE and prevent the esophageal mucosal damage, and validate its therapeutic use in esophageal reflux disease.

Effect of chlorogenic acid on antioxidant activity of Flos Lonicerae extracts

Flos Lonicerae is a medically useful traditional Chinese medicine herb. However, little is known about the antioxidant properties of Flos Lonicerae extracts. Here the antioxidant capacity of water, methanolic and ethanolic extracts prepared from Flos Lonicerae to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and reduce Fe3+ to Fe2+ is examined. Chlorogenic acid, a major component of Flos Lonicerae, is identified and further purified from 70% ethanolic extract with high performance liquid chromatography (HPLC) and its antioxidant capacity is characterized. The total phenolic compounds and chlorogenic acid con-tents in Flos Lonicerae are determined. The present results demonstrate that the Flos Lonicerae extracts exhibit antioxidant ac-tivity and chlorogenic acid is a major contributor to this activity.

Antioxidation Activities of Low-Molecular-Weight Gelatin Hydrolysate Isolated from the Sea Cucumber Stichopus japonicus

Gelatin extracted from the body wall of the sea cucumber （Stichopus japonicus） was hydrolyzed with flavourzyme. Low-molecular-weight gelatin hydrolysate （LMW-GH） of 700-- 1700 Da was produced using an ultrafiltration membrane bioreaetor system. Chemiluminescence analysis revealed that LMW-GH scavenges high free radicals in a concentration-dependent manner; IC50 value for superoxide and hydroxyl radicals was 442 and 285 μgmL-1, respectively. LMW-GH exhibited excellent inhibitory characteristics against melanin synthesis and tyrosinase activity in B16 cells. Furthermore, LMW-GH notably increased in- traeellular glutathione （GSH）, which in turn suppressed melanogenesis. LMW-GH performs antioxidation activity, holding the potential of being used as a valuable ingredient in function foods, cosmetics and pharmaceuticals or nutriceuticals.

Antioxidant activity of chito-oligosaccharides on pancreatic islet cells in streptozotocin-induced diabetes in rats

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.

Tannins from Canarium album with potent antioxidant activity

The contents of total phenolics and extractable condensed tannins in the leaves, twigs and stem bark of Canarium album were determined. The structural heterogeneity of condensed tannins from stem bark was characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS） and nuclear magnetic resonance （NMR） analyses The results show the predominance of signals representative of procyanidins and prodelphinidins. In addition, epicatechin and epigallocatechin polymers with galloylated procyanidin or prodelphinidin were also observed. The tannins were screened for their potential antioxidant activities using l,l-diphenyl-2-picrylhydrazyl （DPPH） and ferric reducing/antioxidant power （FRAP） model systems. Tannins extracted from leaves, twigs and stem bark all showed a very good DPPH radical scavenging activity and ferric reducing power.

Sulfated polysaccharides with antioxidant and anticoagulant activity from the sea cucumber Holothuria fuscogliva

Sea cucumber is a traditional nutritional food and medicinal resource with many bioactive components in China. Holothuriafuscogliva is a big sea cucumber with a rich of bioactive polysaccharides. To investigate the bioactivities of the polysaccharides from sea cucumber H. fuscogliva, we prepared the sulfated polysaccharides （HfP） from sea cucumber H. fuscogliva using a protease hydrolysis method. Antioxidant activities of HfP were investigated, including hydroxyl radical scavenging activity and superoxide radical scavenging activity. And, the anticoagulant activities of HfP were studied, including the activated partial thromboplastin time （APTT）, prothrombin time （PT） and thrombin time （TT）. The average molecular weight was 1 867.1 Da, with a sulfate content of 20.7%. In addition, the molar ratio of monosaccharide composition of HfP was Man： Rha： Glc A： Glc： Gal： Xyl： Fuc=0.083 6： 0.437： 0.134： 0： 1.182： 0.748： 1. It had a strong antioxidant activity, the hydroxyl and superoxide radical scavenging activity ECs0 of HfP was 3.74 and 0.037 mg/mL, respectively. It also showed a good anticoagulant activity in our study. The APTT of HfP was much higher than that of heparin sodium, and the PT and TT of HfP was close to that ofheparin sodium at a low concentration. Therefore, HfP shows a good antioxidant and anticoagulant activity and it may become a potential candidate of the natural antioxidant and anticoagulant and will have a good application future in health product or medicine industry.