Effects of Centella asiatica extracts on apoptosis of mouse spleen lymphocytes

AIM：To isolate the active compounds of Centella asiatica and to investigate their effects on the apoptosis of mouse spleen lymphocytes. METHODS：Cold soak, polar extraction, silica gel column chromatography and Sephadex LH-20 column chromatography were performed to isolate the active compounds from Centella asiatica, which were identified by the assays of scavenging activity on 1,1-diphenyl-2-picrylhydrayl radical (DPPH·) and hydroxyl radical (OH·). Flow cytometry was performed to observe the effects of Centella asiatica extracts on the apoptosis of spleen lymphocytes in mice. RESULTS：Four extracts (B1, B2, B3 and B4) were obtained by the separating procedure. Among them, the extract B4 exerted a potent scavenging effect on both DPPH· and OH·. Quercetin was identified from the extract B4 by HPLC analysis. The mouse spleen lymphocytes with extract B4 treatment showed a significant rise in the level of apoptosis compared with control group. CONCLUSION：Centella asiatica extract B4 induces apoptosis of spleen lymphocytes, and quercetin is the major substance of extract B4.     

Effects of Centella asiatica extracts on cell cycle and intracellular free calcium in spleen lymphocytes

AIM: To investigate the effects of Centella asiatica extracts on the cell cycle and intracellular free calcium ( _i) in spleen lymphocytes. METHODS: Centella asiatica was extracted with alcohol and different organic solvents. Sephadex-LH20 gel column chromatography was applied for further purification. To screen the active constituents, the scavenging ability of the fractions generated by elution of chromatography against radicals of OH·, DPPH· and their anti-proliferative effects on spleen lymphocytes were measured. To assess the cell cycle and _i in spleen lymphocytes, flow cytometry was performed. RESULTS: The D fraction eluted from the crude extract of Centella asiatica was obtained, which had strong suppressive effect on lymphocyte proliferation and decreased the _i of the cells in a dose-dependent manner. The cell cycle of the spleen lymphocytes arrested at S and G₂/M stages when treated with the D fraction. CONCLUSION: The effects of Centella asiatica for lowing _i and arresting cell cycle at S and G₂/M stages of spleen lymphocytes are responsible for its anti-proliferative activitiy.     

不同种类老山芹总酚和总黄酮含量及抗氧化能力的初步研究

以红色老山芹、绿色老山芹、混合色老山芹的叶片和叶柄为试材,测定不同种类、不同部位老山芹的总酚、总黄酮含量及抗氧化能力,并分析功能活性与物质成分含量之间的相关性。结果表明:不同种类、不同部位老山芹的总黄酮含量、总酚含量大体上表现差异显著,3种老山芹叶片的总黄酮含量和总酚含量均显著高于叶柄;红色老山芹叶片醇提物的抗氧化能力最强,其次为混合色老山芹叶片,绿色老山芹叶柄最弱;各处理醇提物浓度在一定范围内与DPPH·自由基、OH·自由基、ABTS+自由基的清除能力均呈线性关系,醇提物浓度与抗氧化能力呈正相关;老山芹醇提物的抗氧化能力与叶片、叶柄总酚含量呈显著或极显著正相关,与总黄酮含量的相关性不显著。     

草莓和蓝莓果实花青素提取及定量方法的比较

 本研究旨在比较果实花青素提取液的提取效果以及花青素分析方法的准确性。用丙酮-水-甲酸（A）、乙腈-乙酸（B）、乙醇-水-乙酸（C）、甲醇-水-乙酸（D）以及含有盐酸的甲醇（E） 等5种提取液分别提取了草莓和蓝莓果实中的花青素,用比色法和液相色谱法分别测定了花青素的含量。试验结果表明：由于提取液中有机溶剂和酸种类的不同,提取效果差异甚大。提取草莓花青素时,以D的提取效果最好,其次是E、C、A,B的提取量最低,还不到D的1/3。提取蓝莓花青素得到的结果与草莓相似。用比色法分析,D、E、C提取液提取的草莓和蓝莓果实的花青素含量明显低于液相色谱分析的结果, 但B提取液比液相色谱结果高2倍以上,A提取液提取蓝莓时比色法与液相色谱法的结果相近,而提取草莓时比色法比液相色谱结果高出约50%。建议花青素的定量方法应尽量使用液相色谱法,对同一种果实,尤其是以低沸点醇类作提取液时可以用比色法。     

Antioxidant Capacity,Phenolic Compounds and Antioxidant Enzymes of Wild Grape Seeds from Different Accessions Grown in Iran

Grape seeds contain high levels of phytochemicals, that have been associated with a reduced risk of chronic diseases. In this study, the polyphenolic content, antioxidant capacity and antioxidant enzymes activity in the seeds of 20 wild grape accessions native to Iran were investigated. The antioxidant capacity of the wild grape seeds was evaluated by DPPH radical scavenging activity, ferric reducing antioxidant power (FRAP) and Fe²⁺ chelating. Total phenolic (TP), total flavanoid (TF) and total proanthocyanidin (TPA) content ranged from 44.86 to 155.1, 23.21 to 131.91, and 10.11 to 26.10?mg/g seed, respectively. All seed extracts showed remarkable DPPH radical scavenging activity, ferric reducing antioxidant power (FRAP) and Fe²⁺ chelating ranging from 27.34 to 78.57%, 198.83 to 590.86?µmol TE/g and 23.12 to 62.98%, respectively. Grape seed extracts had the greatest antioxidant enzymes activity guaiacol peroxidase (G-POD), superoxide dismutase (SOD), and catalase (CAT). Based on HPLC analyses, catechin was the most abundant compound identified (25.89?mg/g) for all accessions. Additionally, significant correlations among different antioxidant assays were found. With the high amount of total phenolic content, antioxidant capacity and antioxidant enzymes activity, seeds of P6, P7 and P8 accessions might be evaluated as a nutritional supplement.     

不同钾盐浸种对无棱丝瓜种子萌发和出苗的影响

研究了不同浓度的3种钾盐对无棱丝瓜种子萌发和出苗的影响。结果表明,浓度在0.4～0.6g/L的硝酸钾、磷酸二氢钾浸种对无棱丝瓜种子发芽率、发芽指数、出苗率和出苗指数有明显的促进作用;0.2～0.4g/L硫酸钾浸种对种子萌发和出苗有一定促进作用,而高于0.4g/L后则有抑制作用。     

两种粤产粉孢牛肝菌抗氧化活性

研究两种产自广东的粉孢牛肝菌(Tylopilus)——类铅紫粉孢牛肝菌(T.plumbeoviolaceoides)和花盖粉孢牛肝菌(GDGM27210)的子实体水提液和75%乙醇提取液的体外抗氧化活性。结果表明,在实验浓度范围内,两种牛肝菌的子实体水提液和75%乙醇提液有不同程度的抑制亚油酸自氧化和清除自由基DPPH.、羟自由基.OH的作用。其中水提液对抑制亚油酸自氧化作用效果较好,最大抑制率分别可达95.74%和92.79%;而75%乙醇提取液对清除DPPH.效果较好,最大清除率分别可达86.68%和94.03%。     

Physico-chemical properties and antioxidant capacity of different jujube (Ziziphus jujuba Mill.) cultivars grown in loess plateau of China

The physico-chemical properties and antioxidant capacity of the five main jujube varieties: Junzao, Lingbaozao, Jinzao, Zanhuangzao and Lizao collected from loess plateau of China were determined. The analyzed components included fruit yield, moisture content, total mass per fruit, pH, titratable acidity (TA), reducing sugars, total sugars, total soluble solids (TSS), ascorbic acid, total phenolics content, total flavonoids content and several pure phenolic compounds. The output of the analyses showed higher amounts of ascorbic acid and phenolics in these jujube varieties than in some common fruits. The antioxidative capacity of the jujube extracts, evaluated with the reducing power, the β-carotene bleaching, the 2,2-diphenyl-1-picrylhydracyl (DPPH•), and the 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonicacid) (ABTS•⁺) scavenging methods, showed that the antioxidant activity of the extracts of Lingbaozao was excellent for free radical scavenging and a potent natural antioxidant of commercial value. Statistically significant differences were observed between jujube cultivars investigated with regards to the measured parameters except rutin content of fruit. These results demonstrated that the cultivar was the main factor which influences the physico-chemical properties and antioxidant activity of jujubes.     

几种香蕉废弃物清除DPPH自由基的作用

测定了几种香蕉废弃物不同溶剂提取物及不同生长期香蕉果实(果皮及果肉)的DPPH自由基清除作用;此外,还对不同材料、不同蕉类品种废弃物的DPPH自由基清除效果进行了比较,以期研究香蕉废弃物的抗氧化活性。结果表明:大部分材料的80%甲醇、80%乙醇、80%丙酮提取物均具有很强的DPPH自由基清除作用,其中80%丙酮提取物的自由基清除作用稍强;所有材料中,嫩苞片及花显示了很强的DPPH自由基清除能力,其IC50值分别为0.6834和0.6853mg/mL,不同材料的80%乙醇提取物的DPPH自由基清除作用强弱顺序依次为:嫩苞片花青色果皮(未成熟)黄色果皮(成熟)老苞片果轴;在整个生长期间,香蕉果皮的自由基清除作用由强变弱,而果肉则与之相反;不同蕉类品种的各种废弃物自由基清除作用强弱顺序依次为:香蕉(威廉斯B6,Williams B6)红蕉大蕉粉蕉。     

HPLC-DAD-MS-DPPH在线筛选与定性黑脉羊肚菌抗氧化活性成分

为建立在线高效液相色谱-高分辨质谱-2,2-二苯代苦味酰基苯肼(high-performance liquid chromatogra phy-diode array detection-mass spectrometry-DPPH,HPLC-DAD-MS-DPPH) 快速筛选和定性黑脉羊肚菌(Morchella angusticeps)中抗氧化活性成分的方法,对黑脉羊肚菌样品的水提物、50%乙醇提取物、95%乙醇提取物和氯仿提取物进行检测,并应用高分辨质谱对筛选所得抗氧化活性成分进行定性分析.结果 显示,黑脉羊肚菌水提物抗氧化活性最优,筛选获得抗氧化活性成分3个,应用高分辨质谱进行定性分析,获得抗氧化成分分别为山梨醇、未知化合物和柠檬酸.故可知,黑脉羊肚菌抗氧化活性成分主要为水溶性小分子极性化合物,以水提取物中含量最为丰富.本方法直观、快速、准确,适用于复杂天然产物中抗氧化活性成分的快速筛选与定性.     

粗毛纤孔菌不同极性萃取物对α-葡萄糖苷酶的抑制作用及抗氧化活性研究

为探讨粗毛纤孔菌(Inonotus hispidus)子实体对α-葡萄糖苷酶抑制作用及体外抗氧化活性的影响,采用系统溶剂萃取法对粗毛纤孔菌甲醇粗提物进行萃取,得到石油醚、乙酸乙酯、正丁醇极性萃取物.选用福林酚比色法、α-葡萄糖苷酶活性抑制法、1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻...     

苍耳有效成分的提取及其杀虫活性的研究

分别用石油醚、苯、丙酮和乙醇4种溶剂提取苍耳植株的有效成分,将获得的4种提取物以不同浓度处理菜粉蝶的幼虫,研究这4种提取物对菜粉蝶的杀虫活性。结果表明:这4种提取物对菜粉蝶都具有生物活性,随着浓度的减小,杀虫活性也随之降低,但石油醚提取物在低浓度情况下具有较高的拒食率。其中,就拒食作用而言,以乙醇提取物的效果最好,48 h不同浓度的提取物(0.08 g/mL和0.32 g/mL)拒食率分别为78.53%和96.24%;就触杀作用而言,以丙酮提取物的效果最好,24、48 h的校正死亡率分别为77.78%和81.48%;而毒杀作用以乙醇的提取物作用最好,1、4、7 d的校正死亡率分别为73.33%、85.33%和92.12%。     

二十种植物与南方根结线虫寄主关系的研究

以繁殖系数Rf为指标,在盆栽条件下采用2龄幼虫接种法研究了7科20种植物与南方根结线虫(Meloidog yne incognita)的寄主关系.结果表明:绿架豆是易感寄主;油菜、翠绿青豆、棱形丝瓜、板蓝根、芥菜、四季精选小香葱、苏丹草和甜玉米是寄主;紫花苜蓿、八棱脆、黄芩、甜高粱、小麦、苇状羊茅、金谷2401、高羊茅是弱寄主,对M.incognita具有一定的抗性;蓖麻、高丹草、黑麦草是非寄主.     

独活提取物抑制柑橘病原真菌活性成分的分离与鉴定

【目的】为了探明独活抑制柑橘病原真菌的活性成分。【方法】采用生长速率法、溶剂萃取、硅胶柱层析、气相色谱-质谱联用(GC-MS)等方法研究独活乙醇提取物对3种柑橘贮藏病害病原真菌的抑菌作用,并分离和鉴定其抑菌活性成分。【结果】独活乙醇提取物中含有高效抑制柑橘炭疽菌的活性物质。GC-MS鉴定B2组分中含有4种香豆素类化合物—异佛手柑内酯、牛防风素、茴芹内酯、异茴芹内酯。异佛手柑内酯、茴芹内酯、异茴芹内酯3种化合物对柑橘炭疽菌的EC50在45 mg.L-1左右,其中茴芹内酯的抑制活性最好,其EC50为41.521 6 mg.L-1。显微观察发现,茴芹内酯处理导致柑橘炭疽菌菌丝体分支增多、畸形,部分隔膜消失等变化。【结论】异佛手柑内酯、茴芹内酯、异茴芹内酯为独活抑制柑橘病原真菌的主要活性物质。     

丝瓜种质资源遗传多样性的SSR与SRAP分析

利用SSR和SRAP标记对30份丝瓜种质资源进行遗传多样性分析,分别从52对SSR和48对SRAP引物中选取10对和12对多态性好的引物,其中10对SSR引物共扩增出32条多态性带,12对SRAP引物扩增出118条多态性带。30份种质间的平均遗传相似系数为0.761,说明丝瓜间种质遗传背景比较狭隘。聚类分析显示,30份丝瓜种质被划分为普通丝瓜与有棱丝瓜两大类。     

Fingerprinting antioxidative activities in plants

Background

A plethora of concurrent cellular activities is mobilised in the adaptation of plants to adverse environmental conditions. This response can be quantified by physiological experiments or metabolic profiling. The intention of this work is to reduce the number of metabolic processes studied to a minimum of relevant parameters with a maximum yield of information. Therefore, we inspected 'summary parameters' characteristic for whole classes of antioxidative metabolites and key enzymes.

Results

Three bioluminescence assays are presented. A horseradish peroxidase-based total antioxidative capacity (TAC) assay is used to probe low molecular weight antioxidants. Peroxidases are quantified by their luminol converting activity (LUPO). Finally, we quantify high molecular weight superoxide anion scavenging activity (SOSA) using coelenterazine. Experiments with Lepidium sativum L. show how salt, drought, cold, and heat influence the antioxidative system represented here by TAC, LUPO, SOSA, catalase, and glutathione reductase (GR). LUPO and SOSA run anti-parallel under all investigated stress conditions suggesting shifts in antioxidative functions rather than formation of antioxidative power. TAC runs in parallel with GR. This indicates that a majority of low molecular weight antioxidants in plants is represented by glutathione.

Conclusion

The set of assays presented here is capable of characterising antioxidative activities in plants. It is inexpensive, quick and reproducible and delivers quantitative data. 'Summary parameters' like TAC, LUPO, and SOSA are quantitative traits which may be promising for implementation in high-throughput screening for robustness of novel mutants, transgenics, or breeds.     

Antidiabetic,Antihypertensive and Antioxidant Properties of Grapevine Leaf Extracts Obtained by Ultrasound,Microwave Assisted,and Classical Solvent Extraction

Grapevine leaves are consumed as a food in Mediterranean countries. At the same time, it is a byproduct of the wine industry. Antioxidant, antihypertensive and antidiabetic properties of grapevine leaves after ultrasound-assisted extraction (UAE), microwave assisted extraction (MAE) and classical solvent extraction (CSE) were evaluated and compared by using water and methanol as solvent. DPPH radical scavenging activity, total phenolic content (TPC), total antioxidant capacity (TEAC), metal chelating activity, ACE inhibitory activity and α?glucosidase inhibitory activity of grapevine leaves extracts were determined. The highest values of grapevine leave extracts for antioxidant properties; DPPH (61.60%), TPC (910.18?mg GAE?g^?1), TEAC (6.23?µg Trolox mg^?1) and metal chelating activity (IC50; 3.18?mg mL^?1) were obtained from UAE when the methanol was used extracting solvent. However antihypertensive activity (13.68%) and antidiabetic activity (83.00%) values obtained from MAE by using water as a solvent were relatively highest. The effectiveness of UAE, MAE and CSE methods in terms of antioxidant, antihypertensive and antidiabetic properties of grapevine leaves were significantly (p?<?0.05) different from each other. This differences depending on the solvent used.     

淹水对红籽瓜嫁接苗生长和根系生理生化的影响

以丝瓜为砧木,"中信1号"红籽瓜为接穗,研究淹水胁迫对红籽瓜嫁接苗和自根苗生长和根系生理生化特性的影响.研究结果表明,淹水胁迫后,嫁接苗的生物量和根系活力均显著高于自根苗.随着淹水胁迫时间的延长,可溶性糖和脯氨酸含量均呈先上升后下降的趋势,嫁接苗植株上升幅度较大而下降幅度较小,并始终显著高于同期的自根苗.嫁接苗根系MDA含量先升后降,而在自根苗中持续上升,且同期自根苗增加幅度明显大于嫁接苗.     

硒在大蒜体内的生物富集及其抗氧化作用

大蒜对硒具有强的生物富集作用，并能将吸收的无机硒的７８．２４％转化为有机硒。经ＳｅｐｈａｄｅｘＧ－１５０柱层析，聚丙烯酰胺凝胶电泳，ＳＤＳ－聚丙烯酰胺凝胶电泳，首次从富硒大蒜中分离到一种分子量为３００００Ｄ的含硒蛋白，进一步研究其抗氧化作用，发现该蛋白能提高谷胱甘肽过氧化物酶、超氧化物歧化酶、过氧化氢酶活性，降低过氧化物酶活性。ＥＰＲ表明，Ｎａ２ＳｅＯ３处理组大蒜体内活性氧相对含量比对照低３１．９１％。     

Effects of EDRV and DIDS on reactive oxygen species levels in acute ischemia-reperfusion injured myocardium

AIM: To investigate the effects of chloride channel inhibitor 4,4- diisothiocyanostilbene-2,2- disulfonic acid (DIDS) and free radical scavenger edaravone(EDRV) on the production of reactive oxygen species in acute ischemia-reperfusion injured (I/RI) myocardium. METHODS: Male Sprague-Dawley rats, subjected to myocardial ischemia for 30 min and reperfusifor for 4 h, were divided into 5 groups: sham group, I/RI group, DIDS group, EDRV group and DIDS+EDRV group. The rats were treated with EDRV (10 mg/kg for 5 min) before reperfusion or/and DIDS (14 mg/kg,4 mL·kg^-1·h^-1 for 2 h) at the beginning of reperfusion by a program-controlled injection micropump . The myocardiac tissues were collected immediately at the end of reperfusion. The levels of ROS, OH· and O₂^- were determined by the methods of spectrofluorophotometry and colorimetry. Myocardial apoptosis was detected by TUNEL method. Serum superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentration were measured by colorimetry. RESULTS: Compared with I/RI group, myocardial apoptotic index, the levels of ROS, O₂^-, OH· and MDA were significantly reduced, and SOD activity was significantly increased in DIDS group, EDRV group and DIDS+EDRV group (P<0.05). Compared with DIDS group, the levels of ROS,O₂^-, OH· and MDA were significantly decreased, and SOD activity was significantly increased in EDRV group (P<0.05).No statistical difference of myocardial apoptotic index between these two groups was observed (P>0.05). Compared with DIDS+ EDRV group, myocardial apoptotic index in DIDS group and EDRV group was significantly increased (P<0.05), and no significant difference of ROS, O₂^-, OH·, MDA and SOD between the two groups was found (P>0.05).CONCLUSION: DIDS and EDRV protect myocardial cells from apoptosis by inhibition of ROS activity. Combinative use of the two reagents has stronger cardioprotectiue effect, suggesting that they have different regulatory pathways.