苦槛蓝叶中的黄酮类化合物

为了解苦槛蓝(Myoporum bontioides A.Gray)的化学成分,采用色谱分离技术从苦槛蓝叶片中分离得到11个黄酮类化合物。通过波谱分析,他们的结构分别鉴定为:桔皮素(1)、甜橙素(2)、5,4′-二羟基-6,7,8,3′-四甲氧基黄酮(3)、4′,5,7,8-四甲氧基黄酮(4)、去甲基川陈皮素(5)、5-羟基-6,7,3′,4′-四甲氧基黄酮(6)、3′,4′,5,6,7,8-六甲氧基黄酮(7)、二氢山柰酚(8)、木犀草素(9)、3′,4′,5,7-四羟基-3-甲氧基黄酮(10)和芹黄素(11)。除化合物9之外,其他化合物均为首次从苦槛蓝叶片中分离得到。孢子萌发法测定结果表明,化合物1,2,8和9对香蕉炭疽菌(Colletotrichum musae)具有较好的抑菌活性。这为苦槛蓝叶片中有效成分的利用提供了理论依据。     

艾纳香化学成分的分离与鉴定

采用色谱技术对艾纳香的化学成分进行分离,根据理化性质和波谱学技术确定化合物的结构。结果从艾纳香的乙酸乙酯萃取液中分离得到7个已知成分,分别鉴定为木犀草素(1);木犀草素-7-甲醚(2);鼠李素(3);5,4′-二羟基-7-甲氧基-黄酮(4);5,4′-二羟基3,7,3′-三甲氧基-黄酮(5);北美圣草素(6);二氢斛皮素-4′-甲醚(7)。其中化合物4、5为首次从该植物中分离得到。     

刺五加有效成分及体外抑制二酰基甘油酰基转移酶活性研究

本文为研究刺五加(Acanthopanax senticosus Harms)的化学成分及其抑制二酰基甘油酰基转移酶(DGAT)活性。刺五加用75%乙醇提取,经硅胶、ODS、半制备HPLC进行分离纯化,结合理化性质、波谱数据鉴定化合物的结构。得到12个化合物分别鉴定为赤式-愈创木基丙三醇-β-O-4′-二羟基松柏醇(1)、(E)-3-(2,2-dimethyl-2H-chromen-6-yl)prop-2-enal(2)、7′E-4,9-dihydroxy-3,3′,5′-trimethoxy-8,4′-oxyneolign-7′-en-9′-al(3)、5-甲氧基去氧双松柏醇(4)、去氧双松柏醇(5)、5,5′-二甲氧基落叶松脂素(6)、5,5′-二甲氧基开环异落叶松树脂酚(7)、(7′S,8′S)-4′-O-甲基黄花菜木脂素(8)、(+)-9′-O-(Z)-阿魏酰-5,5′-二甲氧基落叶松脂素(9)、(+)-9′-O-(E)阿魏酰-5,5′-二甲氧基落叶松脂素(10)、大豆苷(11)和3′-甲氧基大豆苷(12)。其中化合物1～3和8～10为首次从该植物中分离得到。化合物1、3～7、9和10对DGAT1活性具有抑制作用,其IC_(50)值范围在81.5±1.2到123.2±1.1μM之间。     

海南栽培肾茶的化学成分研究

为了解肾茶(Clerodendranthus spicatus)的化学成分,从海南栽培肾茶地上部分分离得到11个化合物,经波谱分析分别鉴定为:吐叶醇(1)、丁香脂素(2)、3,4-二羟基苯乙醇(3)、甜橙素(4)、5,6,7,4′-四甲氧基黄酮(5)、5-羟基-6,7,3′,4′-四甲氧基黄酮(6)、6-羟基-5,7,4′-三甲氧基黄酮(7)、5-羟基-6,7,3′,4′-四甲氧基黄烷酮(8)、3,3′,5-三羟基-4′,7-二甲氧基-二氢黄酮(9)、松脂素(10)和熊果酸(11)。化合物3、9和10为首次从肾茶中分离得到。对化合物1~6进行活性测试,结果表明化合物3~5对乙酰胆碱酯酶具有一定的抑制活性。     

城口东俄芹化学成分研究

采用硅胶、聚酰胺、Sephadex LH-20、大孔吸附树脂等柱色谱方法从城口东俄芹的全草中分离得到6个化学成分,根据理化性质和NMR波谱分析分别鉴定为suberosin(1)、crenulatin(2)、7-羟基-8-甲氧基香豆素(3)、luteolin-7-O-α-L-arabinopyranosyl(1→6)-β-D-glucopyranoside(4)、芹菜素-7-O-β-D-葡萄糖苷(5)和daphnolin(6)。所有化合物均首次从东俄芹属中分离得到,其中3、4和6首次从伞形科中分离得到。     

木荷茎中木脂素类化学成分研究

为了研究山茶科植物木荷(Schima superbaGardn.et Champ.)茎中的化学成分,采用硅胶柱层析、Sepha-dex LH-20、HPLC等色谱方法进行分离,NMR和MS等波谱学方法进行结构鉴定,从木荷茎中分离鉴定了9个木脂素类化合物,分别为丁香脂素(syringaresinol,1)、pinoresinol(2)、落叶松脂醇(lariciresinol,3)、balanophonin(4)、3-acetoxymethyl-5-[(E)-2-formylethen-1-yl]-2-(4-hydroxy-3-methoxyphenyl)-7-methoxy-2,3-dihydrobenzofuran(5)、4,9-dihydroxy-3,5′-dimethoxy-4′,7-epoxy-8,5′-neolignane-9′-yl acetate(6)、二聚松柏醇(dihydrodehydrodico-nifery alcohol,7)、异落叶松脂素(isolariciresinol,8)和1-(3,4-dimethoxyphenyl)-2-O-(2-methoxy-4-omegahydroxypropylphenyl)propane-1,3-diol(9)。以上木脂素类化合物均为首次从该植物分离得到。     

千里香化学成分研究CSCD

研究千里香的化学成分。采用硅胶、ODS、Sephadex LH-20、制备液相等多种色谱分离技术进行分离纯化,通过波谱数据分析进行结构鉴定。从千里香70%乙醇提取物中分离得到26个化合物,分别鉴定为千里香脂素(1)、8-去甲基川陈皮素(2)、ficusal(3)、lariciresinol-4′-monomethy ether(4)、(±)-5′-methoxy-4′-O-methyllariciresinol(5)、diospyrosin(6)、(-)-9′-O-E-feruloyl-lyoniresinol(7)、7-O-methylphellodenol-B(8)、欧芹烯酮酚甲醚(9)、3,4′-二羟基-3′,5′-二甲氧基苯丙酮(10)、4′-羟基-5,7-二甲氧基二氢黄酮(11)、5-羟基-6,7,3′,4′-四甲氧基二氢黄酮(12)、4′-羟基-5,7,3′-三甲氧基二氢黄酮(13)、5,7,3′,4′,5′-五甲氧基二氢黄酮(14)、2′,4-二羟基-3′,4′,6′-三甲氧基查尔酮(15)、2′,3-二羟基-4,4′,6′-三甲氧基查尔酮(16)、楝叶吴萸素B(17)、2′-羟基-3,4,5,4′,6′-五甲氧基查尔酮(18)、2′-羟基-3,4,4′,6′-四甲氧基查尔酮(19)、5,8-二羟基-6,7,3′,4′-四甲氧基黄酮(20)、3′-羟基-5,6,7,8,4′,5′-六甲氧基黄酮(21)、5,3′,5′-三羟基-7,4′-二甲氧基黄酮(22)、5,7,3′-三羟基-8,4′-二甲氧基黄酮(23)、3′-羟基-5,6,7,4′-四甲氧基黄酮(24)、5,7,3′,4′,5′-五甲氧基黄酮(25)、5-羟基-6,7,8,3′,4′-五甲氧基黄酮(26),其中化合物1、2为新化合物,化合物3~8、10~13、15、16、20~24为首次从九里香属植物中分离得到,化合物17为首次从千里香中分离得到。     

幌伞枫叶的化学成分研究

为了解幌伞枫(Heteropanax fragrans)的药理活性化学基础,从其叶的乙醇提取物中分离得到8个化合物,经波谱分析分别鉴定为:(7S,8R)-蛇菰脂醛素-4-O-β-D-吡喃葡萄糖苷(1)、4β,10α-香木兰烷二醇(2)、原儿茶酸(3)、3′-甲氧基-槲皮素-3-O-β-D-吡喃葡萄糖苷(4)、山柰酚-3-O-β-D-吡喃葡萄糖苷(5)、槲皮素-3-O-β-D-吡喃葡萄糖苷(6)、槲皮素-3-O-β-D-芸香糖苷(7)和山柰酚-3-O-β-D-芸香糖苷(8)。这8个化合物均为首次从幌伞枫中分离得到。     

酸豆外果皮黄酮类成分及其抗氧化活性评价

为了研究酸豆外果皮成分及其抗氧化活性,采用现代色谱分离技术从酸豆(Tamarindus indica Linn.)外果皮丙酮提取物中分离和纯化得到18个黄酮类化合物,通过核磁共振波谱数据对它们进行鉴定,分别为木犀草素(1)、7,3′,4′-三羟基黄酮(2)、芹菜素(3)、7,4′-二羟基黄酮(4)、7,4′-二羟基-3′-甲氧基黄酮(5)、紫云英苷(6)、槲皮素(7)、山柰酚(8)、金合欢素(9)、圣草酚(10)、柚皮素(11)、紫铆素(12)、二氢山柰酚(13)、5,7,3′,5′-四羟基二氢黄酮(14)、5,6,7,4′-四羟基二氢黄酮(15)、3,5,7,3′,5′-五羟基二氢黄酮醇(16)、儿茶素(17)、表儿茶素(18)。10个化合物(2、4～6、9、11、13～16)为首次从该属植物中分离得到;采用ORAC法体外抗氧化活性测定,发现酸豆外果皮的4份萃取物及9个化合物(1、2、7～10、12、13、17)有较好的抗氧化活性。     

福建省不同产地及不同生育期土荆芥精油化学成分的比较

采用气相色谱-质谱联用技术(GC-MS),对来源于福建省不同产地(福州新店镇、平潭县城关、连城县城关)以及不同生育期(苗期、生长期、果熟期)土荆芥(Chenopodium ambrosioides L. )地上部分精油的化学成分及其相对含量进行了比较分析.结果表明,福州新店镇、平潭县城关和连城县城关产土荆芥精油的主要化学成分(相对含量0.04%以上)分别有12、11和27种,3个产地的共有成分为11种;福州新店镇产土荆芥精油中相对含量较高的成分有对-聚伞花素(49.60%)、α-松油烯(26.81%)和异驱蛔素(8.16%)等,平潭县城关产精油中相对含量排在前3位的成分也为对-聚伞花素(47.25%)、α-松油烯(27.27%)和异驱蛔素(8.82%),连城县城关产精油中相对含量较高的成分有对-聚伞花素(36.33%)、2,5-二甲基-3-己炔-2,5-二醇(20.03%)、α-松油烯(12.71%)和异驱蛔素(3.07%)等.苗期、生长期和果熟期土荆芥精油的主要成分分别有17、12和12种,3个时期的共有成分为10种;苗期精油中相对含量较高的成分有对-聚伞花素(49.75%)、异驱蛔素(10.77%)、1-(1,3-二甲基-3-环己烯-1-基)乙酮(8.08%)和2,5-二甲基-3-己炔-2,5-二醇(7.79%)等;生长期精油中相对含量较高的成分有对-聚伞花素(49.60%)、α-松油烯(26.81%)和异驱蛔素(8.16%)等;果熟期精油中相对含量较高的成分有对-聚伞花素(43.65%)、α-松油烯(24.09%)和异驱蛔素(19.12%)等.研究结果显示,产地的地理环境和土荆芥的生育期对福建产土荆芥地上部分精油的化学成分及其相对含量均有一定的影响.     

海南国产沉香的化学成分研究

从国产沉香(Aquilaria sinensis)的95%乙醇提取物中分离得到6个化合物,经波谱数据分析,分别鉴定为3,3'-(3-hydroxypropane-1,2-diyl)diphenol(1)、guaiacy lacetone(2)、6-羟基-2-(4'-羟基-2-苯乙基)色酮(3)、6-羟基-2-(2-羟基-2-苯乙基)色酮(4)、6-羟基-2-(2-苯乙基)色酮(5)和5α,6β,7α,8β-四羟基-2-(4'-甲氧基-2-苯乙基)-5,6,7,8-四氢色酮(6)。其中化合物1为新化合物,化合物3为首次从国产沉香中分离得到。     

Cleavage of double-stranded DNA by the intrinsic 3'-5' exonuclease activity of DNA polymerase B1 from the hyperthermophilic archaeon Sulfolobus solfataricus at high temperature

The substrate requirement of the intrinsic 3'-5' exonuclease of DNA polymerase B1 from the hyperthermophilic archaeon Sulfolobus solfataricus P2 (Sso polB1) was investigated. Sso polB1 degraded both single-stranded (ss) and double-stranded (ds) DNA at similar rates in vitro at temperatures of physiological relevance. No difference was found in the cleavage of 3'-recessive, 3'-protruding and blunt-ended DNA duplexes at these temperatures. However, a single-stranded nick in duplex DNA was less readily employed by the enzyme to initiate cleavage than a free 3' end. At lower temperatures, Sso polB1 cleaved ssDNA more efficiently than dsDNA. The strong 3'-5' exonuclease activity of polB1 was inhibited by 50% in the presence of 2 microM dNTPs, but remained measurable at up to 600 microM dNTPs. In view of the strong exonuclease activity of Sso polB1 on matched dsDNA, we suggest that S. solfataricus may have evolved mechanisms to regulate the exonuclease/polymerase ratio of the enzyme, thereby reducing the cost of proofreading at high temperature.     

Anti-CD11d antibody treatment reduces free radical formation and cell death in the injured spinal cord of rats

Treatment with a monoclonal antibody (mAb) against the CD11d subunit of the leukocyte integrin CD11d/CD18 after spinal cord injury (SCI) decreases intraspinal inflammation and oxidative damage, improving neurological function in rats. In this study we tested whether the anti-CD11d mAb treatment reduces intraspinal free radical formation and cell death after SCI. Using clip-compression SCI in rats, reactive oxygen species (ROS) generated in injured spinal cord were detected using 2',7'-dichlorofluorescin-diacetate and hydroethidine as fluorescent probes. ROS in the injured cord increased significantly after SCI; anti-CD11d mAb treatment significantly reduced this ROS formation. Immunohistochemistry and western blotting were employed to assess the effects of anti-CD11d mAb treatment on spinal cord expression of gp91Phox (a subunit of NADPH oxidase producing superoxide) on formation of 4-hydroxynonenal (HNE, indicating lipid peroxidation) and on expression of caspase-3. We also assessed effects on cell death, determined by cell morphology. The expression of gp91Phox, formation of HNE, and cell death increased after SCI. Anti-CD11d mAb treatment clearly attenuated these responses. In conclusion, anti-CD11d mAb treatment significantly reduces intraspinal free radical formation caused by infiltrating leukocytes after SCI, thereby reducing secondary cell death. These effects likely underlie tissue preservation and improved neurological function that result from the mAb treatment.     

β,β''-Iminodipropionitrile Impairs Retrograde Axonal Transport

beta,beta'-Iminodipropionitrile (IDPN), a neurotoxin, causes redistribution of neurofilaments in axons followed by the development of proximal axonal swellings and, in chronic intoxication, a distal decrease in axonal caliber. The latter changes are caused by a selective impairment in the slow anterograde axonal transport of neurofilament proteins. To assess the role of retrograde axonal transport in IDPN toxicity, we used [3H]N-succinimidyl propionate ([3H]NSP) to label covalently endogenous axonal proteins in sciatic nerve of the rat and measured the accumulation of radioactively labeled proteins in the cell bodies of motor and sensory neurons over time. IDPN was injected intraneurally 6 h or intraperitoneally 1 day before subepineurial injection of [3H]NSP into the sciatic nerve, and the animals were killed 1, 2, and 7 days after [3H]NSP injection. Neurotoxicity was assessed by electron microscopic observation of the nerves of similarly treated animals. Both intraneural and intraperitoneal injection of IDPN caused an acute reduction in the amount of labeled proteins transported back to the cell bodies. The early appearance of these changes suggests that alterations in retrograde transport may play a role in the production of the neuropathic changes.     

The targets of 2,2',5,5'-tetrachlorobiphenyl in the respiratory chain of rat liver mitochondria revealed by modular kinetic analysis

The response of the respiratory subsystem of oxidative phosphorylation to the environmental pollutant, 2,2',5,5'-tetrachlorobiphenyl (2,2',5,5'-TCB) was investigated by modular kinetic approach. The effects of 20 M 2,2',5,5'-TCB on the activity of the respiratory chain modules in rat liver mitochondria oxidizing succinate (+ rotenone) in state 3 were assessed. The toxin inhibited the rate of respiration by 23%. Analysis around cytochrome c revealed that 2,2',5,5'-TCB inhibited both cytochrome c-oxidizing and - reducing modules. The toxin inhibited also CoQ-oxidizing module, however it did not affect the kinetics of CoQ-reducing module. Taken together, these data indicated that 2,2',5,5'-TCB inhibited cytochrome bc₁ but had no effect on succinate dehydrogenase.     

Analysis of effects of 2,2',5,5'-tetrachlorobiphenyl on the flux control in oxidative phosphorylation system in rat liver mitochondria

Modular kinetic analysis reveals that the environmental pollutant 2,2',5,5'-tetrachlorobiphenyl (2,2',5,5'-TCB) affects a large number of steps in oxidative phosphorylation in rat liver mitochondria. 2,2',5,5'-TCB increases membrane permeability to ions, and inhibits NADH dehydrogenase, cytochrome bc1, cytochrome oxidase (all in the respiratory chain) and ATP-synthase (in the phosphorylation subsystem). Surprisingly, flux control distribution does not change. A kinetic model for oxidative phosphorylation was used to simulate these findings, and it was found that combined large changes in the processes indicated indeed left the flux control largely unchanged. In addition, computational analysis with the model indicated that the adenine nucleotide translocator might be inhibited by 2,2',5,5'-TCB.     

Biochemical Study of Heterosis for Brain Myelin Content in Mice

Heterosis (hybrid vigor) for brain myelin content has been examined in detail in (C57BL/6J x DBA/2J)F1 hybrid mice at 17 days of age. The amount of myelin isolated from the F1 hybrid brain is greater than that isolated from either parental strain. In addition, the total protein content in the myelin of the three genotypes showed the following trend: F1 greater than DBA greater than C57. However, no discernible differences in myelin protein compositions could be detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Analysis of the whole brain for several myelin-associated constituents such as GM1 ganglioside, 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase), 5'-nucleotidase, and carbonic anhydrase indicated that heterosis exists for these components. No heterosis was found for such nonmyelin constituents as gangliosides GD1a, GT, GQ, RNA, DNA, and choline acetyltransferase. A developmental study of the whole brain CNPase indicated that the heterotic effect was greatest during the most active period of myelination (17-30 days). We conclude that the heterotic effect is specific for myelin content and is probably the result of an accelerated myelin synthesis. The heterotic effect should have great potential as a new model for studying aspects of myelinogenesis.     

Rat Brain Adenosine Deaminase After 2''-Deoxycoformycin Administration: Biochemical Properties and Evidence for Reduced Enzyme Levels Detected by 2''-[3H]Deoxycoformycin Ligand Binding

Near total inhibition of brain adenosine deaminase (ADA) activity in rats injected with the potent ADA inhibitor 2'-deoxycoformycin (DCF) was previously shown to reduce enzyme activity for up to 50 days during which time the enzyme exhibited reduced sensitivity to in vivo inhibition by DCF. Here, we investigated the biochemical properties of ADA and the basis for its reduced activity after DCF treatment. It was found that much higher doses of DCF were required to inhibit ADA in DCF-treated compared with drug-naive animals. Fourteen days after DCF administration, reduced ADA activity in brain homogenates was due to a decrease in Vmax, rather than to an altered Km of ADA for adenosine. DCF treatment had no effect on Ki values for erythro-9-(2-hydroxy-3-nonyl)adenine inhibition of ADA. The IC50 value for DCF inhibition of ADA in hypothalamus was unchanged. However, the Ki for DCF inhibition of ADA in whole brain increased by fivefold. Sucrose gradient analysis of brain ADA revealed only one corresponding peak of activity and [3H]DCF-labeled ADA in DCF-treated and control rats. A radioligand filtration assay with [3H]DCF was developed to assess the effects of DCF on ADA protein levels. Over a roughly 200-fold range of ADA activities the binding of [3H]DCF was highly correlated with deaminase activity (r = 0.99). In brain tissues taken 8 and 33 days after treatment of rats with DCF, [3H]DCF binding was reduced to 27% and 48% of control levels, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibition of M and P Phenol Sulfotransferase by Analogues of 3''-Phosphoadenosine-5''-Phosphosulfate

Structural analogues of the sulfate donor 3'-phosphoadenosine-5'-phosphosulfate (3',5'-PAPS) were examined for their ability to inhibit dopamine and phenol sulfation by the M and P forms of phenol sulfotransferase (PST), respectively. The Ki values for each of the adenosine derivatives were calculated from the rate equation for PST. For both M and P PST, the naturally occurring product 3'-phosphoadenosine-5'-phosphate, (3',5'-PAP), was shown to be the most effective inhibitor. The weakest inhibitors of the two sulfotransferases were 5'-adenosine phosphosulfate and the three AMP derivatives, which were less than 1,000 times as effective as 3',5'-PAP. 5'-ATP, 2',5'-PAPS, 2',5'-PAP, and 5'-ADP were similar in their inhibition of M and P PST and were all approximately 100 times less effective than the natural end product. These data reveal that there is a rigid structural requirement for binding of the ribose portion of adenosine to both M and P PST that involves the groups on both the 3' and 5' positions. The effectiveness of binding to the two enzymes may depend on both steric factors as well as the distribution of negative charges on the ribose ring.     

Effect of 5''-(N-Ethylcarboxamido)adenosine on Adenosine Transport in Cultured Chromaffin Cells

Extracellular adenosine is transported into chromaffin cells by a high-affinity transport system. The action of adenosine receptor ligands was studied in this cellular model. 5'-(N-Ethylcarboxamido)adenosine (NECA), an agonist of A2 receptors, activated adenosine transport. Km values for adenosine were 4.6 +/- 1.0 (n = 5) and 10.2 +/- 3.0 microM (n = 5) for controls and 100 nM NECA, respectively. The Vmax values were 66.7 +/- 23.5 and 170.2 +/- 30 pmol/10(6) cells/min for controls and 100 nM NECA, respectively. The A1 agonist N6-cyclohexyladenosine, the A1 antagonist 8-cyclopentyl-1, 3-dipropylxanthine, and the A1-A2 antagonist 1,3-dipropyl-8-(4-[(2-aminoethyl)amino]-carbonylmethyloxyphenyl)- xanthine did not significantly modify the adenosine transport in this system. Binding studies done with [3H]dipyridamole, a nucleoside transporter ligand, did not show changes in either the number or affinity of transporter sites after NECA treatment. This ligand can enter cells and quantifies the total number of transporters. The binding studies with [3H]-nitrobenzylthioinosine, which quantifies the plasma membrane transporters, showed a Bmax of 19,200 +/- 800 and 23,200 +/- 700 transporters/cell for controls and 100 nM NECA, respectively. No changes in the KD were obtained. The effects of NECA were not mediated through adenylate cyclase activation, because its action was not imitated by forskolin.