不同基质栽培的药用鲍姆桑黄孔菌体外抗肿瘤活性的比较研究

比较3种不同栽培技术的鲍姆桑黄孔菌Sanghuangporus baumii——3年生栎树段木鲍姆桑黄孔菌、桑枝代料鲍姆桑黄孔菌和发酵鲍姆桑黄孔菌中多糖、总酚、黄酮及总三萜含量和体外抗肿瘤活性。分别采用硫酸-蒽酮、福林-酚、亚硝酸-硝酸铝和香草醛-冰醋酸法对4种主要成分进行含量测定。选用人大细胞肺癌细胞（H460）、人前列腺癌细胞（PC3）、人乳腺癌细胞（MDA-MB-231）、人肝癌细胞（SMMC-7721和BEL-7402）和人神经母细胞瘤细胞（SH-SY5Y）为待检细胞株,采用CCK-8检测细胞活力。结果显示,发酵鲍姆桑黄孔菌多糖含量最高（29.64%）,代料鲍姆桑黄孔菌和3年生段木鲍姆桑黄孔菌多糖含量均低于2.57%;2种代料鲍姆桑黄孔菌的总酚和黄酮含量较高,而发酵鲍姆桑黄孔菌最低（0.24%和0.68%）,尤其是代料鲍姆桑黄孔菌1号的总酚和黄酮含量高达4.02%和32.13%;三萜含量在4种栽培法的鲍姆桑黄孔菌中差异不明显。体外对6种肿瘤细胞增殖抑制能力最强的均为代料鲍姆桑黄孔菌,其IC₅₀值最低,其次是3年生段木鲍姆桑黄孔菌,发酵鲍姆桑黄孔菌在本研究设置的浓度范围内无明显的细胞毒性。结果表明：不同栽培方式鲍姆桑黄孔菌产生体外抗肿瘤活性不同,代料鲍姆桑黄孔菌最强,其次是3年生段木鲍姆桑黄孔菌,发酵鲍姆桑黄孔菌活性最差,这种活性差异与其有效成分含量高低直接相关;鲍姆桑黄孔菌发挥抗肿瘤活性的物质基础主要是总酚和黄酮。     

鲍姆桑黄孔菌子实体HPLC指纹图谱及抗氧化活性谱效关系

本研究对38个不同批次和菌龄的椴木栽培鲍姆桑黄孔菌Sanghuangporus baumii子实体醇提物进行了高效液相色谱（HPLC）分析,建立了S. baumii醇提物HPLC指纹图谱,提取得到27个共有峰信息（P1-P27）;同时,对38个样品醇提取的Trolox等价抗氧化能力（TEAC）进行分析,基于正交偏最小二乘（OPLS）回归建立了抗氧化活性预测模型,潜变量数为3,交互验证所得累积模型预测参数（Q²）为0.803,变量累积解释指标（R²X）和（R²Y）分别为0.574和0.954,相关系数平方（R²）为0.954,经验证模型可以获得良好的预测效果,可以准确反映谱峰与活性之间相关关系;利用模型变量投影重要性（VIP）指标筛选得到14个共有峰对抗氧化活性具有显著贡献。研究获得的鲍姆桑黄孔菌抗氧化谱效关系模型为桑黄原料活性的预测提供了有效的方法,可辅助指导鲍姆桑黄孔菌活性化合物的分离纯化,也为鲍姆桑黄孔菌栽培子实体的品质控制提供了技术手段。     

粗毛纤孔菌的化学成分及抗肿瘤活性成分

本文研究了粗毛纤孔菌的化学成分及抗肿瘤活性成分。对粗毛纤孔菌的甲醇提取物进行石油醚、乙酸乙酯、正丁醇萃取,采用Sephadex LH-20凝胶色谱法,反相C18柱色谱法及高效液相色谱法对不同萃取组分进行分离纯化。分离得到8个化合物,经鉴定分别为麦角甾醇、齿孔酸、4-(3,4-二羟苯基)-3-丁烯-2-酮、phellibaumin A、3,3?-亚甲基双[6-[2-(3,4-二羟苯基)乙烯基]-4-羟基-2H-吡喃-2-酮](MBP)、肌苷、原儿茶酸和原儿茶醛。其中化合物MBP为首次从自然界中分离得到,对其进行了MTT抗肿瘤筛选和细胞凋亡分析。结果表明此化合物对人肝癌细胞HepG2的细胞增殖具有抑制作用,IC50值为2.3μg/mL,并且可以诱导HepG2细胞凋亡,且呈一定的剂量依赖关系。本研究明确了MBP的提取方法,初步断定该化合物抗肿瘤活性是通过诱导细胞凋亡实现的。     

青藏高原黑果枸杞花青素对人肝癌细胞增殖和自噬的影响

探究黑果枸杞花青素在体外对人肝癌HepG2细胞增殖和自噬的影响。利用CCK-8法测定细胞活力,EdU和细胞划痕试验检测细胞增殖和迁移效果,RT-PCR和Western blot检测增殖和自噬相关基因的mRNA和蛋白表达。结果显示,黑果枸杞花青素可有效抑制人肝癌HepG2细胞的增殖和迁移;上调增殖因子(LATS1、LATS2和MOB1)和自噬因子(Beclin-1、LC3-Ⅱ和AMPK),并下调增殖因子YAP的mRNA水平;下调自噬因子p-mTOR和细胞周期因子CDK4,并上调自噬因子p-AMPK和LC3-Ⅱ的蛋白表达。由此推测黑果枸杞花青素可在体外抑制人肝癌HepG2细胞增殖和迁移,并促进人肝癌HepG2细胞发生自噬。     

原儿茶酸与原儿茶醛对小鼠肝脏细胞色素P450表达的影响

目的:研究原儿茶酸和原儿茶醛对小鼠肝脏细胞色素P450酶主要亚型表达水平的影响。方法:将C57BL/6小鼠随机分为正常对照组(0.9%生理盐水)、原儿茶酸与原儿茶醛给药组(6 mg/kg),苯巴比妥诱导组(80mg/kg),每24 h给药一次,连续给药2周。提取肝脏中总RNA,RT-PCR技术考察药物对CYP450主要亚型mRNA表达的影响;制备肝微粒体,用Western blot技术考察P450酶主要亚型的表达。结果:RT-PCR结果显示,在mRNA水平上,与正常对照组相比,原儿茶酸可诱导Cyp1a2基因的表达(P0.05),对Cyp2c37、Cyp2d9的mRNA表达无诱导作用。原儿茶醛对Cyp1a2的mRNA的表达有诱导作用(P0.05),对Cyp2c37和Cyp2d9的mRNA的表达无诱导作用。Western blot结果显示原儿茶酸对CYP1A2,CYP2E1表达有显著诱导作用,对其他几种亚型无影响。原儿茶醛对CYP1A2表达有显著诱导作用,对其他几种亚型无影响。Western blot结果与RT-PCR结果基本一致。结论:原儿茶酸与原儿茶醛均可以显著诱导CYP1A2的mRNA以及蛋白表达,并且原儿茶酸可以显著诱导CYP2E1的蛋白表达。     

三株亚肉座菌乙酸乙酯提取物生物活性及GC-MS分析

采用乙酸乙酯提取3株亚肉座菌菌丝体,测试虫生真菌乙酸乙酯提取物（EAE）的抗肿瘤、抗菌和抗氧化活性,并借助GC-MS方法分析各提取物中的化学成分。结果发现2株亚肉座菌的菌丝体EAE对HepG2细胞的抑制活性较强,IC₅₀均小于9μmol/L;抗菌结果表明2株真菌的提取物具有抑制细菌生长的作用;1株供试菌的EAE表现出较强的DPPH自由基清除活性（清除率可达85%）。GC-MS分析表明从亚肉座菌JXJG201717、JXJG201720和ARSEF7697的EAE中分别鉴定出21、35和39种成分,主要成分为酯类、醇类和酸类;盘状亚肉座菌JXJG201720与ARSEF7697有相同化合物13个,与暹罗亚肉座菌JXJG201717存在7个相同化合物。本研究表明虫生亚肉座菌具有产生丰富活性成分的能力,彰显出多种利用价值。     

NF-kB 拮抗剂PDTC 对HepG2 细胞的抑制及对caspase-3 表达的影响

目的:研究NF-kB拮抗剂PDTC诱导人肝癌HepG2细胞凋亡及其对凋亡相关基因caspase-3表达的影响,初步探讨其诱导凋亡的可能机制。方法:以不同浓度的PDTC处理人肝癌HepG2细胞,利用MTT法检测对人肝癌HepG2细胞凋亡的影响;利用RT-PCR和Western-blot检测caspase-3mRNA和蛋白的表达。结果:不同浓度PDTC作用人肝癌HepG2细胞不同时间后,能够显著抑制HepG2细胞的生长增殖,存在剂量和时间依赖性(P<0.05);PDTC能够上调HepG2细胞中caspase-3mRNA和蛋白的表达。结论:NF-kB拮抗剂PDTC对人肝癌HepG2细胞产生显著抑制,并能上调HepG2细胞中caspase-3mRNA和蛋白的表达。     

NF-kB拮抗剂PDTC对HepG2细胞的抑制及对caspase-3表达的影响

目的：研究NF-kB拮抗剂PDTC诱导人肝癌HepG2细胞凋亡及其对凋亡相关基因caspase-3表达的影响,初步探讨其诱导凋亡的可能机制。方法：以不同浓度的PDTC处理人肝癌HepG2细胞,利用MTT法检测对人肝癌HepG2细胞凋亡的影响;利用RT-PCR和Western-blot检测caspase-3mRNA和蛋白的表达。结果：不同浓度PDTC作用人肝癌HepG2细胞不同时间后,能够显著抑制HepG2细胞的生长增殖,存在剂量和时间依赖性（P〈0.05）;PDTC能够上调HepG2细胞中caspase-3mRNA和蛋白的表达。结论：NF-kB拮抗剂PDTC对人肝癌HepG2细胞产生显著抑制,并能上调HepG2细胞中caspase-3mRNA和蛋白的表达。     

几种天然有机多酚酸体外抗脂质过氧化研究

通过天然有机多酚酸对H2O2诱导小鼠肝匀浆脂质过氧化和自发脂质过氧化的保护作用的体外实验,比较研究绿原酸、咖啡酸、迷迭香酸、阿魏酸、丹参素、原儿茶醛和3-羟基肉桂酸体外抗氧化能力的强弱,探讨其抗氧化能力产生机理及构效关系。结果表明,7种多酚酸对H2O2诱导小鼠肝匀浆脂质过氧化的保护作用强弱依次为迷迭香酸咖啡酸原儿茶醛绿原酸阿魏酸丹参素3-羟基肉桂酸,其最大和最小抑制率相差2.4倍;对小鼠肝脏组织自发脂质过氧化的抑制率大小顺序为迷迭香酸、原儿茶醛、绿原酸、咖啡酸、阿魏酸、丹参素、3-羟基肉桂酸,最高和最低抑制率相差近90%。7种天然有机酸抗脂质过氧化作用因化合物结构不同存在显著差异。     

Synthesis and evaluation of bifunctional nitrocatechol inhibitors of pig liver catechol-O-methyltransferase

Bifunctional compounds were tested in vitro as potential inhibitors of pig liver catechol-O-methyltransferase (COMT) with respect to the catechol substrate 4-[(3,4-dihydroxyphenyl)azo]benzenesulfonate. The bifunctional compounds were a composite of either two nitrocatechols or one nitrocatechol and one phenol, linked by amide bonds to a spacer unit comprising two to five methylene groups. The unsymmetrical compounds N-[2-(4-hydroxybenzoylamine)ethyl]-3,4-dihydroxy-5-nitrobenzamide], N-[3-(4-hydroxybenzoyl-amine)propyl]-3,4-dihydroxy-5-nitrobenzamide] and N-[5-(4-hydroxybenzoylamine)pentyl]-3,4-dihydroxy-5-nitrobenzamide] demonstrated strong inhibitory action against COMT with K(i) values in the 100 nM range. In comparison, the monofunctional nitrocatechol analogues of these compounds had K(i) values that were significantly higher.     

Studies on the Compounds Produced by Molds

Three isocoumarin compounds (BV 1, 2 and 3) were isolated from the cultural broth of Aspergillus oniki 1784. BV 1 was mellein (3-methyl-8-hydroxy-3,4-dihydroisocoumarin). BV 2 and 3 were assigned to be 3-methyl-4,8-dihydroxy-3,4-dihydroisocoumarin, 3-methyl-3,8-dihydroxy-3,4-dihydroisocoumarin, respectively. These two compounds (BV 2, 3) were newly isolated. Also another component named BV 4 was proved to be 6-methylsalicylic acid.     

A New Antioxidant Cyclic Diarylheptanoid from the Rhizomes of Zingiber officinale

The bioassay was directed to the fractionation of the methanol extract of the rhizomes of Zingiber officinale Roscoe leading to the isolation of a new antioxidant cyclic diarylheptanoid. The structure of the new compoundwas established as 1,5-epoxy-3-hydroxy-1-(3,4-dihydroxy-5-methoxy-phenyl)-7-(3,4-dihydroxy-phenyl) heptane (1) on the basis of MS, 1D and 2D-NMR experiments.     

The formation of dihydrodiols by the chemical or enzymic oxidation of benz[a] anthracene and 7,12-dimethylbenz[a] anthracene.

When benz[a] anthracene was oxidised in a reaction mixture containing ascorbic acid, ferrous sulphate and EDTA, the non-K-region dihydrodiols, trans-1,2-dihydro-1,2-dihydroxybenz[a] anthracene and trans-3,4-dihydro-3,4-dihydroxybenz[a] anthracene together with small amounts of the 8,9- and 10,11-dihydrodiols were formed. When oxidised in a similar system, 7,12-dimethylbenz[a] anthracene yielded the K-region dihydrodiol, trans-5,6-dihydro-5,6-dihydroxy-7,12-dimethylbenz[a] anthracene and the non-K-region dihydrodiols, trans-3,4-dihydro-3,4-dihydroxy-7,12-dimethylbenz[a] anthracene, trans-8,9-dihydro-8,9-dihydroxy-7,12-dimethylbenz[a] anthracene, trans-10,11-dihydro-10,11-dihydroxy-7,12-dimethylbenz[a] anthracene and a trace of the 1,2-dihydrodiol. The structures and sterochemistry of the dihydrodiols were established by comparisons of their UV spectra and chromatographic characteristics using HPLC with those of authentic compounds or, when no authentic compounds were available, by UV, NMR and mass spectral analysis. An examination by HPLC of the dihydrodiols formed in the metabolism, by rat-liver microsomal fractions, of benz[a] anthracene and 7,12-dimethylbenz[a] anthracene was carried out. The metabolic dihydriols were identified by comparisons of their chromatographic and UV or fluorescence spectral characteristics with compounds of known structures. The principle metabolic dihydriols formed from both benz[a] anthracene and 7,12-dimethylbenz[a] anthracene were the trans-5,6- and trans-8,9-dihydrodiols. The 1,2- and 10,11-dihydrodiols were identified as minor products of the metabolism of benz [a] anthracene and the tentative identification of the trans-3,4-dihydriol as a metabolite was made from fluorescence and chromatographic data. The minor metabolic dihydriols formed from 7,12-dimethylbenz[a] anthracene were the trans-3,4-dihydrodiol and the trans-10,11-dihydriol but the trans-1,2-dihydrodiol was not detected in the present study.     

Identification of Potential Bioactive Compounds from Aspergillus terreus against HCV NS3 Serine Protease

This study was aimed to isolate bioactive compounds from the fermentation products of Aspergillus terreus, which could inhibit NS3 protease of hepatitis C virus (HCV). The bioactive compounds were isolated by reverse-phase silica-gel column chromatography, semi-preparative reversed-phase, and Sephadex LH-20, and then their structures were elucidated through spectroscopic analysis. As a result, two small molecule compounds were isolated. Compound 1 was identified as a new benzaldehyde, (E)-2,4-dihydroxy-6-propenylbenzaldehyde. Compound 2 was identified as pleurone, which was obtained from microorganisms for the first time. Their inhibitory activities against HCV NS3 protease (IC₅₀) were 32.6 μM and 78.9 μM, respectively. This study provided a new option for the development of anti-HCV drugs.     

Structures and synthesis of the growth inhibitors batatasins IV and V,and their physiological activities

Two new compounds, batatasins IV and V were isolated from dormant bulbils of Chinese yam (Dioscorea batatas) and shown to be 2′,3-dihydroxy-5-methoxybibenzyl and 2′-hydroxy-3,4,5-trimethoxybibenzyl, respectively. An analogue, 3,4′-dihydroxy-5-methoxybibenzyl was synthesized. Inhibitory activities of these three compounds as well as batatasin I (6-hydroxy-2,4,7-trimethoxyphenanthrene) and batatasin III (3,3′-dihydroxy-5-methoxy-bibenzyl) in lettuce seed germination, lettuce hypocotyl elongation and wheat coleoptile section elongation tests are described.     

Isolation of novel phenolic compounds with multidrug resistance (MDR) reversal properties from Onychium japonicum

We isolated seven novel compounds, namely, 3',4',6-trihydroxy-2,4-dimethoxy-3-(3″,4″-dihydroxybenzyl)chalcone (1), 3',6-dihydroxy-2,4,4'-trimethoxy-3-(3″,4″-dihydroxybenzyl)chalcone (2), α,β-dihydro-3',6-dihydroxy-2,4,6'-trimethoxy-3-(3″,4″-dihydroxybenzyl)chalcone (3), 3',4,4'-trihydroxy-2,6-dimethoxychalcone (4), 4',5,7-trihydroxy-6-(3″,4″-dihydroxybenzyl)flavone (5), 3-(3',4'-dihydroxybenzyl)-6,7-dihydroxycoumarin (6), 3-(3',4'-dihydroxyphenyl)-3,4-dihydroisocoumarin (7), as well as a known compound, 3',4',7-trihydroxy-5-methoxyflavanone (8) from the whole grass of Onychium japonicum, and elucidated their structures by spectroscopic methods. Compounds 1-3 exhibited significant multidrug resistance (MDR) reversal effects on MCF-7/ADR and Bel-7402/5-Fu cell lines.     

1-(3,4-Dihydroxy-5-Methoxyphenyl-3-Methylbut-2-Ene from The Liverworth Plagiochila Rutilans

Extraction of the liverwort Plagiochila rutilans afforded 1-(3,4-dihydroxy-5-methoxyphenyl)3-methylbut-2-ene, the structure of which was confirmed by synthesis.     

Biotransformation of 3-methylphthalate by Micrococcus sp. strain 12B

When Micrococcus strain 12B grown on o-phthalate was incubated with 3-methylphthalate, three compounds accumulated. These were shown to be 2-pyrone-3-methyl-4,6-dicarboxylic acid, 3,4-dihydroxy-6-methylphthalic acid, and 5-hydroxy-3-methyphthalic acid, all previously undescribed. A pathway for the formation of these compounds is proposed.     

Xanthones from Tovomita pyrifolium

The wood of Tovomita pyrifolium (Guttiferae) contains the novel tovopyrifolins A [1,6-dihydroxy-7-methoxy-5-prenyl-6′,6′-dimethylpyrano (2′,3′:3,2)xanthone], B (1,5-dihydroxy-3,4-dimethoxyxanthone) and C (1,3,5-trihydroxy-2-methoxyxanthone) and also the known tovophyllins A and B [structure revised to 1,6-dihydroxy-5-prenyl-6′, 6′-dimethylpyrano(2′,3′:3,2)-6″,6″-dimethylpyrano(2″,3″:7,8)xanthone].