北虫草不同极性部位体外抗肿瘤活性研究

目的研究山东北虫草不同极性部位体外抗肿瘤活性。方法采用系统溶剂法,分别用石油醚、乙酸乙酯、正丁醇和蒸馏水依次提取北虫草不同极性部位。以乳腺癌MCF-7细胞、食管癌ECA-109细胞和肺癌A549细胞为研究对象,用MTT法检测北虫草不同极性部位对MCF-7细胞、ECA-109细胞和A549细胞的抑制作用。结果石油醚部位在100μg/ml浓度时,对MCF-7细胞、ECA-109细胞和A549细胞的抑制率较强,乙酸乙酯部位在50,100μg/ml浓度对ECA-109细胞的抑制率较强。结论北虫草石油醚部位和乙酸乙酯部位体外抗肿瘤活性较强,值得进一步研究其化学成分。     

菱角壳分级萃取物对肿瘤细胞增殖及凋亡的影响

目的:制备菱角壳乙醇提取物的分级萃取物,并研究其对肿瘤细胞增殖及凋亡的作用。方法:采用有机溶剂萃取法将菱角壳乙醇提取物分为乙酸乙酯萃取物和正丁醇萃取物2个不同极性部分,并测定其黄酮与多酚含量,CCK-8法测定菱角壳分级萃取物对肿瘤细胞增殖的抑制作用,流式细胞术检测菱角壳萃取物诱导肿瘤细胞凋亡的作用。结果:乙酸乙酯萃取物中多酚和黄酮含量高于正丁醇萃取物。在两种萃取物浓度为25、50、100、200、400 μg/mL时均能抑制胃癌细胞(SGC-7901)、肝癌细胞(Hep G2)的增殖,最高抑制率均达60%以上,并存在明显的剂量与效应关系。两种萃取物均能促进肿瘤细胞凋亡,400 μg/mL的乙酸乙酯、正丁醇萃取物作用肝癌细胞24 h,凋亡率分别为45.57%和33.34%;作用胃癌细胞24 h,凋亡率分别为31.01%和21.65%。结论:菱角壳乙酸乙酯萃取物具有抑制肿瘤细胞增殖及促肿瘤细胞凋亡的生物活性。     

荞麦芽提取物抗突变和抑制肿瘤作用的实验研究

对荞麦芽提取物的抗突变和抗肿瘤作用进行研究,为开发荞麦的保健作用提供依据.实验以小鼠骨髓细胞微核实验观察荞麦芽提取物的抗突变作用,以小鼠S-180移植性肿瘤观察荞麦芽提取物的抗肿瘤效果.结果显示,荞麦芽提取物对N-甲基-N-硝基-N-亚硝基胍(MNNG)诱发的小鼠骨髓细胞微核的发生有明显的抑制效果,其中荞麦芽乙醇提取物、乙酸乙酯提取物、正丁醇提取物在80mg/kg度下,对小鼠骨髓微核的抑制率达73.4％、75.8％、74.2％;对小鼠S-180移植性肿瘤生长也有明显的抑制作用,其中荞麦芽乙醇提取物、乙酸乙酯提取物的抑瘤率达39.5％、38.0％.说明荞麦芽对体细胞DNA损伤有保护作用,对肿瘤也有一定的预防作用.     

灵巴菌质抗肿瘤活性部位的体外筛选

通过体外细胞培养对"灵巴菌质"(巴豆发酵品)的石油醚部位和碱性醇提物的抗肿瘤活性进行初步筛选。对"灵巴菌质"以石油醚、二氯甲烷、水饱和正丁醇和甲醇四个极性梯度依次进行抽提,得到石油醚部位(PEE)等4个提取部位;甲醇提取物再采用酸溶碱沉-溶剂萃取等方法处理,得到其碱性醇提物(AME)。通过快速、灵敏的MTT比色法考察石油醚部位和碱性醇提物对SMMC-7721、MGC-803、A549、Hep G-2肿瘤细胞增殖的影响。PEE、AME与巴豆苷(Cro)一样,对A549细胞增殖均有显著抑制作用,且呈浓度依赖性;而二者对MGC-803细胞活力没有明显影响。在一定浓度范围内,AME仅对SMMC-7721细胞增殖有抑制作用;PEE和AME对Hep G-2细胞增殖却有促进作用。"灵巴菌质"的石油醚部位和碱性醇提物对SMMC-7721、MGC-803、A549、Hep G-2等4种瘤株增殖的影响是有一定选择性的,二者仅对A549细胞生长均呈现出明显的抑制作用。     

荞麦芽的抗氧化活性研究

为了研究荞麦芽的抗氧化活性,以不同生长阶段的荞麦芽为研究对象,采用70%乙醇为溶剂提取了不同生长阶段甜荞麦芽和苦荞麦芽的总黄酮,利用比色法测定了荞麦芽提取物对DPPH自由基的清除率。结果表明,不同品种的荞麦芽苗均有一定的抗氧化能力,且苦荞的抗氧化能力明显高于甜荞。并在第14 d时荞麦芽提取物对DPPH自由基的清除能力最高,苦荞麦芽达到80%以上,甜荞麦芽在50%以上,以后几天略有下降。     

黄秋葵花提取物不同极性部位抗氧化活性的研究

为确定黄秋葵花提取物的抗氧化活性,采用有机溶剂萃取法将黄秋葵花提取物分为乙酸乙酯相和水相2个不同极性部位,测定不同极性部位提取物中总黄酮及多酚的含量,分析其还原能力、(1,1-diphenyl-2-picrylhydrazyl)DPPH自由基清除能力、超氧阴离子自由基清除能力和羟自由基清除能力,比较黄秋葵花不同极性部位提取物的抗氧化作用。结果表明,黄秋葵花提取物的不同极性部位均有抗氧化活性作用,其中乙酸乙酯相部位提取物的羟自由基清除能力和超氧阴离子清除能力均强于水相提取部位,而水相提取部位的还原力和DPPH自由基清除能力强于乙酸乙酯相部位提取物。本研究将为黄秋葵花抗氧化功能食品的研究与开发提供理论依据。     

柿叶提取物体外抑瘤作用初步研究

目的:研究柿叶提取物在体外对肿瘤细胞的抑制作用。方法:采用不同方法提取柿子叶有效部位,用MTT法检测其对Hela(人宫颈癌细胞)和H22(鼠肝癌细胞)两种细胞的生长抑制率,并用SPSS进行统计分析。结果:柿叶水、醇提取物作用于Hela细胞,具有明显的抑瘤效应;柿叶水、醇提取物作用于H22细胞,有明显的抑瘤效应。加药后肿瘤细胞在形态以及生长方式上面发生明显改变。结论:柿叶不同提取物在一定浓度下对Hela细胞和H22细胞均有明显的抑制增殖作用。     

平盖灵芝提取物对肺腺癌A549细胞增殖抑制作用研究

目的探讨平盖灵芝不同极性部位提取物对肺腺癌A549细胞增殖的抑制作用。方法采用噻唑蓝（MTT）法测定。结果平盖灵芝氯仿部位提取物高剂量组(100μg/mL)对肺腺癌A549细胞的增殖抑制率为30.43%,与空白组比较差异有统计学意义（P<0.01）。结论平盖灵芝氯仿部位提取物对肺腺癌A549细胞增殖有一定抑制作用,其有效成分及作用机制有待进一步研究。     

孜然乙醇提取物的工艺优化及对酪氨酸酶活性的抑制作用

以孜然为原料,在单因素实验的基础上,以酪氨酸酶抑制率为指标,采用响应面法优化孜然乙醇提取物的提取工艺,并测定不同极性部位对酪氨酸酶的影响及孜然乙醇提取物中主要活性成分的含量。结果表明,孜然乙醇提取物抑制酪氨酸酶的最优提取工艺为:乙醇体积分数71%,液料比16:1 (mL/g),提取时间2.2 h,提取温度83 ℃,在此条件下其对酪氨酸酶活性的抑制率达(42.61%±0.56%),与预测值43.20%接近,表明响应面分析的结果可靠;孜然乙醇提取物中多酚、黄酮、皂苷、多糖四种活性成分的含量分别为(1.35%±0.03%)、(4.40%±0.29%)、(0.76%±0.04%)、(4.89%±0.28%);孜然乙醇提取物不同极性部位对酪氨酸酶均有抑制作用,且呈浓度依赖关系,其IC₅₀大小顺序为:水相 > 石油醚相 > 正丁醇相 > 乙酸乙酯相。因此,优化得到的孜然乙醇提取物抑制酪氨酸酶的提取工艺合理、可行,乙酸乙酯和正丁醇部位对酪氨酸酶活性的抑制能力较强,多酚、黄酮、皂苷、多糖等都可能是孜然乙醇提取物抑制酪氨酸酶的功效成分。     

牡丹籽壳提取物及不同极性部位的抗氧化活性研究

采用不同极性溶剂(石油醚、乙酸乙酯、正丁醇以及水)将牡丹籽壳提取物(70%乙醇溶液提取物)逐级系统分离,获得不同极性部位的提取物。研究牡丹籽壳提取物和各极性部位提取物的总多酚含量;采用DPPH及FRAP法,以L-抗坏血酸(VC)和2,6-二叔丁基-4-甲基苯酚(BHT)为参照抗氧化剂考察其抗氧化活性。结果表明:总多酚含量有较大差异,从小到大依次为水部位正丁醇部位乙酸乙酯部位牡丹籽壳提取物,变化范围为1.30~57.50 mg/g;牡丹籽壳提取物与不同极性部位提取物均具有一定的抗氧化能力,总抗氧化活性从小到大依次为正丁醇部位水部位牡丹籽壳提取物乙酸乙酯部位,FRAP值范围为27.28~1 250.08 mmol/L,其中VC和BHT的FRAP值分别为1 814.88 mmol/L和226.00 mmol/L;对DPPH自由基清除能力从弱到强依次为牡丹籽壳提取物乙酸乙酯部位正丁醇部位水部位,半数抑制浓度(IC_(50))范围为22.47~71.00μg/m L,其中BHT的IC_(50)为34.30μg/m L。     

Improving the antioxidant activity of buckwheat (Fagopyrum tataricm Gaertn) sprout with trace element water

Trace element water (TEW) (100, 200, 300, 400 and 500 ppm) was used to grow buckwheat (Fagopyrum tataricm Gaertn) to evaluate whether the beneficial effects of trace elements on the antioxidant activity could be accomplished with the supplement of TEW. At 300 ppm, TEW significantly increased the Cu, Zn and Fe contents in buckwheat sprout, but not the Se and Mn contents. The levels of rutin, quercitrin and quercetin did not differ between buckwheat sprouts grown in TEW and de-ionized water (DIW). The ethanolic extract from buckwheat sprout grown in 300 ppm TEW showed higher DPPH radical scavenging activity, ferrous ion chelating activity, superoxide anion scavenging activity and inhibitory activity toward lipid peroxidation than that grown in DIW. The extract of the TEW group also enhanced intracellular superoxide dismutase activity and resulted in lower level of reactive oxygen species in human Hep G2 cells.     

不同品种甜荞芽中酚类成分的比较

本文建立了甜荞芽5种酚类成分的测定方法,并且对7种甜荞芽中酚类成分进行比较分析。采用超高效液相色谱(UPLC)法检测甜荞中5种多酚成分含量,色谱条件为ACQUITYUPLC? BEHC₁₈色谱柱(2.1 mm×50 mm,1.7 μm),流动相甲醇(A)-0.20%冰乙酸水溶液(B),检测波长为254 nm,流速为0.20 mL/min,进样量为1 μL;并采用高效液相色谱法对甜荞芽中的矢车菊素-3-O-葡萄糖苷和矢车菊素-3-O-芸香糖苷成分的含量进行检测,用DPS 14.50软件对甜荞芽第7、10 d酚类成分的含量进行分析。结果表明,建立的超高效液相色谱法准确可靠,甜荞芽第7、10 d酚类成分总含量具有显著性差异(P<0.05),其中第7、10 d荭草素、异荭草素、牡荆素、异牡荆素、芦丁总含量最高的品种分别是赤甜1号、品甜2号,分别为43.66、40.82 mg/g,含量最低的品种分别是宁荞1号、定甜3号,分别为20.45、14.13 mg/g;矢车菊素-3-O-葡萄糖苷和矢车菊素-3-O-芸香糖苷总含量最高的分别是红花甜荞、宁甜1号,分别为218.47、282.95 mg/kg,最低的分别是赤甜1号、丰甜1号,分别为49.91、88.35 mg/kg。本实验为甜荞芽质量评价提供了依据,为后续研究奠定了基础。     

昆布多糖不同提取工艺优化及其理化性质和抗肿瘤活性比较

在单因素试验的基础上通过正交试验对昆布多糖的提取条件进行优化,并对比分析热水浸提法、超声辅助提取法和复合酶法3 种方法提取的昆布多糖的理化性质和抗肿瘤活性。结果表明,优化后昆布多糖的提取条件为,热水浸提法：提取温度85 ℃、提取时间5 h、料液比1∶80（g/mL）,此条件下多糖平均提取率为5.97%;超声辅助提取法：超声功率500 W、超声时间10 min、料液比1∶90（g/mL）,此条件下多糖平均提取率为5.81%;复合酶法：木瓜蛋白酶75 U/g、果胶酶15 000 U/g、纤维素酶108 000 U/g、pH 5、温度50 ℃、料液比1∶70（g/mL）、时间3 h,此条件下多糖平均提取率为14.29%。不同提取方法对昆布多糖的成分有显著影响,复合酶法提取的昆布多糖的溶解性显著优于水浴和超声提取。提取的昆布多糖中均具有少量蛋白质或多肽存在,并且具有三股螺旋结构。噻唑蓝（MTT）实验表明,不同提取方法得到的昆布多糖具有不同的抗肿瘤活性,其中超声辅助提取多糖对人肺癌细胞A549的抑制作用最强,而复合酶法所得多糖对人乳腺癌细胞MCF-7的抑制效果最好。     

Antimicrobial and human cancer cell cytotoxic effect of synthetic angiotensin-converting enzyme (ACE) inhibitory peptides

Four peptides with high angiotensin-converting enzyme (ACE) inhibitory effect were separated from beef sarcoplasmic protein hydrolysates using commercial enzymes. They were identified as GFHI, DFHING, FHG, and GLSDGEWQ and their 50% inhibition concentration (IC₅₀) values against ACE were 117, 64.3, 52.9, and 50.5 μg/ml, respectively. These peptides were synthesised and further biological activities of these four peptides were measured, including antimicrobial, cytotoxic effect against cancer cells, and macrophage-stimulating effect. Peptide GLSDGEWQ showed growth inhibition on Salmonella Typhimurium, Bacillus cereus, Escherichia coli, and Listeria monocytogenes at a 100 ppm level but not on Staphylococcus aureus and Pseudomonas aeruginosa. Peptide GFHI showed higher inhibition activity on the growth of E. coli and P. aeruginosa at concentrations of 200 and 400 μg/ml. However, peptide FHG inhibited only P. aeruginosa at 200 and 400 μg/ml. The effect of separated peptides on breast cancer (MCF-7), lung cancer (A549), and stomach cancer (AGS) cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Peptide GFHI showed a slight decrease of MCF-7 cell viability in a dose dependent manner. When 400 μg/ml of peptide GFHI was applied to the AGS cell, its viability was decreased by 75%. However, peptide DFHINQ seemed to act as a nutrient to AGS cell because it increased its viability. None of the four peptides had a cytotoxic effect on A549 cells. Nitric oxide (NO) production of peptide GFHI by stimulation of macrophage was investigated at 100, 300, and 1000 μg/ml concentration. NO was not produced in all treatments. From these results it is expected that the ACE inhibitory peptides identified from beef sarcoplasmic protein hydrolysates have both antimicrobial and cancer cell cytotoxic effects.     

Cytotoxicity of Coprinopsis atramentaria extract,organic acids and their synthesized methylated and glucuronate derivatives

Coprinopsis atramentaria is a wild edible mushroom whose methanolic extract revealed a marked antioxidant activity; p-hydroxybenzoic (HA), p-coumaric (CoA) and cinnamic (CA) acids were identified in the extract. In the present work, the cytotoxicity of C. atramentaria extract, previously identified organic acids and their synthesized derivatives (methylated compounds and protected glucuronides) was evaluated. Among all the tested cell lines (MCF-7—breast adenocarcinoma, NCI-H460—non-small cell lung carcinoma, HCT15—colon carcinoma, HeLa—cervical carcinoma and HepG2—hepatocellular carcinoma), the extract presented good activity for the first three cell lines mentioned; in most of the cases methylated and glucuronide derivatives showed a higher activity than the corresponding parental compounds. The substitution of the carboxylic group (in parental organic acids) for an ester (in methylated derivatives) increased the cytotoxicity for tumor cell lines. Acetylated glucuronide derivatives showed higher cytotoxicity when compared with the corresponding parental acids.     

Hairy root extract of Phyllanthus amarus induces apoptotic cell death in human breast cancer cells

This study deals with establishment of hairy root cultures of Phyllanthus amarus using Agrobacterium rhizogenes and cytotoxic effects of methanolic extract of hairy roots on human adenocarcinoma cell line, MCF-7. The hairy root extract displayed a linear concentration- and time-dependent cytotoxicity. Further, increased concentration of the root extract showed an increase in the percent apoptotic cells from 26% to 36% as determined by annexin V-FITC and propidium iodide. The observed cytotoxicity correlated well with the increased levels of intracellular reactive oxygen species (ROS) as well as decreased mitochondrial membrane potential (MMP). The overall results amply suggest an appreciable antiproliferative effect of P. amarus hairy root extract on the MCF-7 cells through induction of apoptosis, thereby establishing the potential anticancer activity of the extract.Industrial relevanceHairy roots of Phyllanthus amarus showing anti cancer properties has great potential as Industrial product.     

蜂胶醇提物对人肝癌细胞Hep G2增殖及凋亡的影响

为探讨蜂胶醇提物对人肝癌细胞Hep G2 增殖和凋亡的影响,采用MTT 法检测蜂胶醇提物对Hep G2 细胞增殖的抑制作用,用荧光倒置显微镜和流式细胞仪分析蜂胶醇提物对Hep G2 细胞凋亡的影响。结果表明：12.5～200μg/mL 质量浓度的蜂胶醇提物作用24、48h 后,均能不同程度地抑制Hep G2 细胞的增殖,呈明显的时间、剂量依赖关系,半数抑制质量浓度(IC50)分别是115、78μg/mL。作用8h 后,Hep G2 细胞出现不同程度的凋亡,凋亡率由6.36% 上升到21.9%,呈现出剂量依赖关系。故蜂胶醇提物可显著抑制人肝癌细胞Hep G2 的生长,诱导其凋亡。     

苦荞萌发物中生物活性黄酮对人乳腺癌细胞增殖的抑制作用

以萌发期(1～6d)的苦荞芽粉为原料,采用铝盐显色可见分光光度法和HPLC 法,探明萌发期(1～6d)苦荞芽粉乙醇提取物中总黄酮、芦丁和槲皮素含量的变化趋势。以人乳腺癌细胞MCF-7 为模型,采用噻唑蓝(MTT)比色法,比较萌发期(1～6d)的苦荞芽粉乙醇提取物对人乳腺癌细胞体外的增殖抑制率。结果表明：苦荞芽粉乙醇提取物具有抑制MCF-7 乳腺癌细胞增殖的作用,尤以萌发第3 天(芦丁与槲皮素含量比为0.92:1)时抑制效果最好,显示二者具有良好的协同抑制效果;苦荞芽粉乙醇提取物的抑制效果与槲皮素和芦丁标准品模拟样品抑制效果相似,表明苦荞芽粉乙醇提取物对MCF-7 细胞的生长起抑制作用的主要功效成分为槲皮素和芦丁。     

Comparison of phenolic compositions between common and tartary buckwheat (Fagopyrum) sprouts

The phenolic compositions of non-germinated/germinated seeds and seed sprouts (at 6–10 day-old) of common (Fagopyrum esculentum Möench) and tartary (Fagopyrum tataricum Gaertn.) buckwheats were investigated. Phenolic compounds, including chlorogenic acid, four C-glycosylflavones (orientin, isoorientin vitexin, isovitexin), rutin and quercetin, were determined in the seed sprouts by high-performance liquid chromatography (HPLC). In the edible parts of common buckwheat sprouts, individual phenolics significantly increased during sprout growth from 6 to 10 days after sowing (DAS), whereas in tartary buckwheat sprouts they did not. While the sum contents of phenolic compounds in the edible part (mean 24.4 mg/g DW at 6–10 DAS) of tartary buckwheat sprouts were similar to those of common buckwheat sprouts, rutin contents in the non-germinated/germinated seeds (mean 14.7 mg/g DW) and edible parts (mean 21.8 mg/g DW) of tartary buckwheat were 49- and 5-fold, respectively, higher than those of common buckwheat. Extracts of the edible parts of both species showed very similar free radical-scavenging activities (mean 1.7 μmol trolox eq/g DW), suggesting that the overall antioxidative activity might be affected by the combination of identified phenolics and unidentified (minor) components. Therefore, buckwheat seed sprouts are recommended for their high antioxidative activity, as well as being an excellent dietary source of phenolic compounds, particularly tartary buckwheat sprouts, being rich in rutin.