基于对接的植物激素3D-QSAR和分子动力学模拟

采用分子对接和分子动力学(MD)模拟方法研究植物雌激素类化合物与雌激素受体的相互作用机制,对接结果表明,雌激素受体活性位点的疏水和氢键作用是影响植物雌激素化合物活性的主要原因,植物雌激素类化合物主要与氨基酸残基GLU353、ARG394、HIS524和LEU525之间形成氢键.然后以对接后的分子构象进行分子结构叠合,结合比较分子力场分析(CoMFA)和比较分子相似性指数分析(CoMSIA)方法建立了3D-QSAR模型.CoMFA模型的交叉验证相关系数(Q2)和非交叉验证相关系数(R2)值分别为0.676和0.994,标准估计误差SEE和F统计量分别为0.143和342.115;CoMSIA模型的Q2=0.565,R2=0.972,SEE=0.286和F=111.480.结果表明,CoMFA和CoMSIA模型具有良好的稳定性和预测能力,可为植物雌激素的雌激素效应研究提供有力的支持.采用MD模拟研究了小分子和受体蛋白的动力学情况,为对接结果的合理性提供了验证.     

多溴联苯醚和全氟烷基酸的分子毒理机制研究

多溴联苯醚(PBDEs)和全氟烷基酸(PFAAs)是两类使用量大、环境污染广泛、人体暴露严重的新型有机污染物,2009年已纳入《斯德哥尔摩公约》持久性有机污染物(POPs)名单,但其毒性效应及作用机制并不明确.本文综述了本课题组近几年针对多溴联苯醚PBDEs和全氟烷基酸PFAAs的分子毒理机制研究工作,主要集中在这两类污染物对甲状腺系统、雌激素系统和肝脏脂肪酸代谢系统干扰效应的分子机制研究.本文分别从分子、细胞和活体三个层面,研究了污染物与核受体的直接结合作用、结合后受体的构象变化、细胞内受体的转录活性、以及活体暴露后受体调控基因的表达变化,由此阐明了污染物通过与受体直接作用导致细胞和活体生物功能改变的分子机制.同时结合计算模拟,探讨了污染物生物效应与其化学结构之间的关系,发现污染物的受体活性取决于它们与受体结合的空间构型,而其活性强度基本与二者的结合能力一致,主要受疏水作用和氢键的影响.此外,还通过研究污染物与天然配体转运蛋白的相互作用,明确了各个污染物与转运蛋白的结合能力,探讨了其构效关系,并评估了污染物对天然配体在体内转运过程的潜在干扰效应.通过上述研究工作,提出了多层面、多靶点研究环境污染物分子毒理机制的新思路,建立和引进了研究污染物与生物靶分子相互作用的新方法,发现了PBDEs、PFAAs与TR、ER、PPARγ核受体结合的新模式,为深入了解这些污染物的分子毒理机制提供了有用的信息和有效的研究手段.     

羟基化多溴联苯醚(OH-PBDEs)在小鼠肝脏微粒体的体外代谢及对CYP450酶活性的影响

羟基化多溴联苯醚(OH-PBDEs)是一类具有内分泌干扰效应的酚类化合物,生物毒性要高于母体多溴联苯醚(PBDEs),研究OH-PBDEs的体外代谢行为对于理解其在生物体内的富集转化具有重要意义。以小鼠肝脏微粒体作为研究对象,考察了3-OH-BDE-47、5-OH-BDE-47、6-OH-BDE-47和2’-OH-BDE-68在小鼠肝脏中的体外代谢,并分别研究了浓度为0.1、0.2、0.4、0.6、1.0μmol·L-1条件下4种OH-PBDEs对细胞色素P450酶系中7-乙氧基香豆素-O-脱乙基酶(ECOD)、7-乙氧基异吩唑酮-O-脱乙基酶(EROD)和苯胺4-羟基化酶(ANH)活性的影响。结果表明,4种OH-PBDEs在小鼠肝脏微粒体中均能够快速代谢,代谢率分别为80%(3-OH-BDE-47)、42%(5-OH-BDE-47)、86%(6-OH-BDE-47)和63%(2’-OH-BDE-68)。实验所设OH-PBDEs各浓度对微粒体的ECOD活性无显著性抑制作用,但对EROD的活性均表现出相同的显著抑制作用;4种OH-PBDEs表现出不同的ANH活性影响,即3-OH-BDE-47对ANH活性具有抑制作用,5-OH-BDE-47具有诱导作用,而6-OH-BDE-47和2’-OH-BDE-68对ANH活性无显著性影响。     

羟基取代和甲氧基取代多溴联苯醚的存在特征和源解析研究进展

近年来,多溴联苯醚（PBDEs）的衍生物,羟基取代的PBDEs（OH-PBDEs）和甲氧基取代的PBDEs（MeO-PBDEs）作为新型的持久性有机污染物引起了国内外学者的关注。这两类物质在各种生物体内（藻类、鱼、贝类、海豹、海鸟及人体血液和乳汁等）和环境介质中（地表水、瀑布和雪）都被检出。OH-PBDEs对生物体具有多种毒性效应,包括干扰甲状腺荷尔蒙动态平衡,干扰氧化磷酸化作用,影响雌二醇合成,并能引起神经毒性效应。而研究表明,在生物体内MeO-PBDEs可通过去甲基化作用转化为OH-PBDEs,因此对生物体内高含量的MeO-PBDEs应予以更密切的关注。OH-PBDEs既有人为源（来自PBDEs生物代谢或自由基反应）又有天然源（在海洋环境中合成）,MeO-PBDEs主要来自天然源。在我国相关的研究刚刚起步。文章比较系统地总结了OH-PBDEs和MeO-PBDEs在环境中存在特征、生物毒性、来源及二者相互关系的研究进展,以期为在我国相关研究的开展提供参考。对于开展关于OH-PBDEs和MeO-PBDEs在特定区域存在特征和源解析的研究,建议重点考察物质的组成和含量与水温、盐度和富营养化程度等因素的关系,以及特定组分的季节性变化,从物质的存在形态和相关关系角度对OH-PBDEs和MeO-PBDEs的来源进行解析。     

典型羟基化多溴联苯醚(OH-PBDEs)小鼠肝微粒体的体外代谢及关键代谢CYP450酶系的研究

羟基化多溴联苯醚(OH-PBDEs)是一类具有内分泌干扰效应的酚类化合物,本研究以小鼠肝脏微粒体作为研究对象,考察了4种典型OH-PBDEs(3-OH-BDE-47、5-OH-BDE-47、6-OH-BDE-47和2′-OH-BDE-68)的体外代谢.研究结果表明,4种OH-PBDEs在小鼠肝脏微粒体中均能被代谢;OH-PBDE结构上的醚键(O)、羟基(OH)和溴原子(Br)的相对位置会影响其代谢效率,OH位于苯环上醚键邻位更有利于其代谢,即6-OH-BDE-47表现出相对较高的代谢率;抑制剂实验发现CYP1A2和CYP3A4是4种典型OH-PBDEs主要的CYP450代谢亚型.     

荧光光谱法研究木栓酮与牛血清白蛋白的相互作用

为了解三萜类化合物在体内的结合转运机制,采用荧光光谱法结合紫外吸收光谱研究从中药爵床中分离的有效成分木栓酮(Friedelin,FDL)与牛血清白蛋白(Bovine serum albumin,BSA)的结合反应.通过研究木栓酮对牛血清白蛋白内源性荧光的淬灭作用,发现木栓酮可以与牛血清白蛋白相互结合,且其作用是一个静态猝灭的过程.通过计算木栓酮与牛血清白蛋白的结合常数和结合位点,发现在生理弱碱性溶液中二者具有较强的结合作用,且以1:1结合.根据热力学参数推测木栓酮与牛血清白蛋白的作用力类型为氢键和范德华力.根据F rster非辐射能量转移机理,计算出木栓酮与牛血清白蛋白结合距离为2.86 nm.研究表明木栓酮在体内可以通过血清白蛋白进行结合和转运.     

苯砜基乙酸酯类化合物发光菌毒性效应的CoMFA研究

应用比较分子场分析方法(CoMFA)对56种苯砜基乙酸酯类化合物的发光菌急性毒性进行了研究,获得了6组分三维定量结构-活性相关模型(3D-QSAR)．模型的交叉验证相关系数(q^2)和常规相关系数(r^2)分别为0．823和0．958,说明该模型具有较高的稳定性和较好的预测能力．模型分析表明：该类化合物发光菌急性毒性效应与化合物分子的空间场和静电场具有显著相关关系．     

分子对接法研究多溴二苯醚衍生物与芳香化酶的作用机理

应用分子对接方法探讨了多溴二苯醚(PBDE)衍生物与人胎盘芳香化酶间的可能作用机理.通过分析对接生成的构象发现,羟基多溴二苯醚(HO-PBDEs)和甲氧基多溴二苯醚(Me O-PBDEs)与人胎盘芳香化酶间形成氢键的情况不同.对人胎盘芳香化酶有抑制活性的HO-PBDEs通过其羟基(—OH)与氨基酸残基Arg435、Arg115、Arg375、Asp309和Ala306形成氢键,而对人胎盘芳香化酶无抑制活性的Me O-PBDEs的甲氧基(Me O—)几乎不与芳香化酶内氨基酸残基形成氢键,这可能是二者对芳香化酶抑制活性不同的原因.由此推测,化合物作为氢键供体可能是多溴二苯醚衍生物具有抑制芳香化酶活性的结构基础.本研究从化合物结构及化合物与人胎盘芳香化酶结合的构象特征上解释化合物活性不同的原因,以期能够利用构象分析得到的结果进行筛选.     

十溴联苯醚（BDE-209）对成年大鼠甲状腺激素的影响

多溴联苯醚(PBDEs)是现代工业中广泛应用的溴系阻燃剂,PBDEs具有持久性有机卤素污染物(Organohalogen Contaminants,OHCs)和内分泌干扰物(Endocrine Disrupters,EDs)的化学物理特性.PBDEs的生物毒性也是国际上环境生态科学的关注热点.论文建立了十溴联苯醚(BDE-209)暴露剂量的成年大鼠模型,测定了不同暴露剂量的实验动物血清甲状腺激素(Thyroid hormones,THs)水平,初步研究了PBDEs污染物对生物体甲状腺激素的影响.BDE-209的量-效关系研究结果显示,TT4、FT4和TT3浓度均随着暴露剂量的提高而下降,这可能是BDE-209对甲状腺合成分泌T4、T3起到了抑制作用.不同BDE-209暴露剂量下,TT4~FT4以及TT4~TT3的正相关系数大于FT3~TT3以及FT3~FT4,这意味着BDE-209暴露对于T4从甲状腺激素转运结合蛋白(Transthyretin,TTR)的分离与脱碘过程影响不明显,而影响了结合态T3从TTR的分离过程以及FT4的脱碘过程,从而对甲状腺激素的平衡具有干扰效应.采用基准点分析法,研究了BDE-209对于甲状腺干扰相对效应的时间-效应关系,实验表明由于甲状腺的应激作用,对于毒物具有自我防御功能,BDE-209对甲状腺的干扰效应在5~8d后才有所显现,持续时间至少为14d.     

三氯酚暴露致稀有鮈鲫肝脏损伤的比较蛋白质组研究

为探讨2,4,6-三氯酚(2,4,6-TCP)暴露致鱼类肝脏损伤的毒性效应,以稀有鮈鲫(Gobiocypris rarus)为模式生物,TCP暴露浓度为1、10、100μg·L-1,进行流水暴露实验(28d),利用二维凝胶电泳方法(2-DE),结合质谱分析技术(MALDI-TOF-MS),对TCP致稀有鮈鲫肝脏损伤的蛋白表达谱进行了研究.结果显示,共发现35个差异蛋白点,其中11个蛋白被成功鉴定(C.I.>95%).对鉴定的蛋白根据GOTerms进行功能分类,发现它们主要参与脂类代谢与转运、氧化应激以及蛋白修复和氧化磷酸化等生物学过程。以上结果表明,TCP对稀有鮈鲫具有明显的肝脏毒性效应.     

取代苯酚类化合物抑藻活性的CoMFA模型

基于比较分子力场分析(CoMFA)方法建立20种取代苯酚类化合物抑藻活性(pI)的三维定量构效关系(3D-QSAR)。训练集中16个化合物用于建立预测模型,测试集5个化合物(含模板分子)作为模型验证。已建立的Co MFA模型的交叉验证系数(Q2)、非交叉验证系数(R2)分别为0.915、0.963,说明所建模型具有较强的稳定性和良好的预测能力。该模型中立体场、静电场贡献率依次为48.4%、51.6%,表明影响抑藻活性(pI)的主要因素是取代基的电荷分布,其次是取代基的疏水性和空间位阻。     

Effects of pulse current electromagnetic fields on human peripheral lymphocytes

The effects of pulsed current electromagnetic fields (PCEMF) on human peripheral lymphocytes sister chromatid exchanges (SCE), and micronuclei (MN) frequencies were investigated. PCEMF were produced by 8/20 μs pulsed current generation with two model types, a long straight wire and a solenoid. Human peripheral lymphocyte samples were exposed to PCEMF (a pulse per min, and continuing for 1 h). SCE and MN assays were used to evaluate the genotoxic effects. The results indicated that SCE and MN incidences correlated positively with B _m and dB/dt. For the long straight wire model, as B _m reached 8.48 × 10^?2 T and dB/dt = 1.06 × 10⁴ T s^?1, the effect was significant. As B _m reached 1.70 × 10^?1 T and dB/dt = 2.13 × 10⁴ T s^?1, the effect was marked. For the solenoid model, the conditions of significant effect were B _m = 0.264 T and dB/dt = 3.30 × 10³ T s^?1 and B _m = 1.32 T and dB/dt = 1.650 × 10⁴ T s^?1. Data indicated that dB/dt was an important factor in increasing the frequencies of MN and SCE in human peripheral lymphocytes.     

Fungicidal activity and molecular modeling of fusarubin analogues from Fusarium oxysporum

Abstract

Fusarubin analogues of Fusarium oxysporum f. sp. ciceris were investigated for antifungal activity in vitro against five soil borne phytopathogenic fungi. 3-O-Methyl-8-O-methyl-fusarubin was inhibitory towards S. sclerotiorum (EC₅₀ 0.33?mmol L^?1) and Sclerotium rolfsii (EC₅₀ 0.38?mmol L^?1). A structure–antifungal activity relationship of fusarubin analogues was established from their activity performance. Possible mechanism of action of these compounds was studied using molecular docking and simulations against three target enzymes which revealed receptor ligand binding affinity. Docking of 3-O-methyl-8-O-methyl-fusarubin into the succinate dehydrogenase site revealed formation of salt bridge, hydrogen bond, π–anion, π–alkyl, and Van der Waals interactions.     

Molecular studies on the ouabain binding site of the Na+, K+-ATPase in milkweed butterflies

Summary. The Na⁺, K⁺-ATPase of the Monarch butterfly (Danaus plexippus) is insensitive to the inhibition by cardiac glycosides due to an amino acid replacement: histidine instead of asparagine at position 122 of the α-subunit representing the ouabain binding site. By PCR amplification of the DNA sequence of this site, a PCR product of 270 bp was obtained from DNA extracted from Danainae species (Danaus plexippus, D. chrysippus, D. gillipus, D. philene, D. genutia, Tirumala hamata, Euploea spp., Parantica weiskei, P. melusine), Sphingidae (Daphnis nerii) and mimics of milkweed butterflies (Hypolimnas missipus, Limenitis archippus and L. arthemis, Nymphalidae). Analysis of the nucleotide sequences revealed that the single point mutation in the ouabain binding domain (AAC-Asn for CAC-His) was present only in Danaus plexippus, but not in the other species investigated. Since these milkweed butterflies also store cardenolides, other structural modifications of the Na⁺, K⁺-ATPase may have occurred or other strategies of cardenolide tolerance have been developed. Received 15 May 2000; accepted 29 June 2000     

Comparative sensitivity of a tropical mysid Metamysidopsis insularis and the temperate species Americamysis bahia to six toxicants

The sensitivity of the tropical mysid, Metamysidopsis insularis relative to that of the sub-tropical North American mysid, Americamysis bahia was determined by comparing their acute toxicity to six toxicants. The 96 h LC50 values for M. insularis ranged from 0.03 mg L^?1 (cadmium (Cd) chloride) to 466 mg L^?1 (potassium chloride (KCL) whereas those for A. bahia ranged from 0.1 mg L^?1 for Cd chloride to 501 mg L^?1 for KCl. The interspecies correlation indicated that the mean acute toxicities for M. insularis showed a high positive correlation with A. bahia (r ² = 1.0). Data suggest there were no significant differences between acute toxic responses of the two species to these toxicants. The results suggest that though M. insularis and A. bahia occur in different climatic zones, the difference in relative sensitivities of the two species due to temperature preferenda may be negligible. Consequently, M. insularis may be considered suitable tropical test species for use in toxicity testing in Trinidad and Tobago.     

A novel extraction method for analysing available sulfamethoxazole in soil

A novel extraction method was established to determine the water-extractable (available) content of sulfamethoxazole (SMX) in soil. The SMX imprinted polymers (MIPs) were synthesised and the performance was evaluated by Fourier transform infrared spectroscopy, scanning electron microscopy and binding experiments. Results showed that the MIPs exhibited good selectivity for SMX, so the MIPs were applied as a sorbent. SMX in soil was extracted by water, sorbed from the extract to MIPs and analysed with a high performance liquid chromatography (HPLC) after its desorption from MIPs. Meanwhile, the classic organic solvent extraction was employed to measure the total SMX content in soil. Results showed that when SMX level in spiked soils varying from 1.0–500?μg?kg^?1, the observed recoveries of available SMX contents ranged from 63.27?±?3.11% to 82.11?±?2.77% (n?=?3), while the total SMX varied between 89.59?±?1.65% and 97.64?±?3.92% (n?=?3). The detection limit of the developed method for SMX in soils was 0.05?μg?kg^?1. Available SMX contents in five field soil samples ranged from 0.13 to 4.14?μg?kg^?1, which were only 0.35–25.40% of the respective total SMX contents. Results from this study manifest the importance of the extents of SMX immobilisation with different soils for assessing SMX's ecological and human health risks.     

有机化学品生物富集因子定量结构-活性关系模型

依据经济合作与发展组织(OECD)关于定量结构-活性关系(QSAR)模型构建和使用导则,将780个有机化合物,以4:1的比例随机划分为训练集(624个化合物)和验证集(156个化合物),通过多元线性回归(MLR)方法构建了一个包含12个描述符的有机化合物鱼类生物富集因子(BCF)的QSAR模型。QSAR模型的调整决定系数R2ad j=0.809,去一法交叉验证系数Q2LOO=0.803,外部验证系数Q2EXT=0.732,表明模型具有较好的拟合优度、稳健性和预测能力。采用欧几里德距离方法表征模型应用域,通过威廉姆斯图分析模型离群点,并对模型进行机理解释。所构建的模型,可以用于预测应用域内有机化学品的生物富集因子。     

Photosynthetic performance of giant clams, Tridacna maxima and T. squamosa, Red Sea

Two species of giant clams, Tridacna maxima and T. squamosa, coexist in the Red Sea, but exhibit distinctly different depth distributions: T. maxima mostly occurs in shallow waters (reef flat and edge), while T. squamosa may occur down to the lower fore-reef slope. Giant clams have been described as mixotrophic, capable of both filter-feeding and photosynthesis due to algal symbionts (zooxanthellae), therefore, observed depth preferences were investigated in relation to possible differences in autotrophy vs. heterotrophy. This study was conducted from April to June 2004, at the reef near the Marine Science Station, Aqaba, Gulf of Aqaba, Red Sea, and in May 2007, at a reef near Dahab, Sinai Peninsula, Egypt. In situ measurements using a submersible pulse amplitude modulated fluorometer (Diving PAM), revealed no significant differences in effective PSII quantum yield (ΔF/Fm′) and relative electron transport rates (ETR) between the two species; but rapid light curves (ETR vs. light, photosynthetically active irradiance, PAR) showed significant differences in maximum photosynthetic rates (ETR_max), with 20% higher values in T. maxima. Chamber incubations displayed higher net and gross oxygen production by T. maxima (88.0 and 120.3 μmol O₂ cm⁻² mantle area day⁻¹) than T. squamosa (56.7 and 84.8 μmol O₂ cm⁻² mantle area day⁻¹); even under shading conditions (simulated depth of 20 m) T. maxima still achieved 93% of the surface gross O₂ production, whereas T. squamosa reached only 44%. A correlation was found between ETR and net photosynthesis measured as oxygen production (T. maxima: R ² = 0.53; T. squamosa: R ² = 0.61). Calculated compensation depth (CD) (gross photosynthesis equals respiration) in T. maxima (16 m) matches the maximum depth of occurrence in this study (17 m). By contrast, the CD of T. squamosa (9 m) was much shallower than the maximum vertical range (42 m). Findings suggest T. maxima is a strict functional photoautotroph limited by light, whereas T. squamosa is a mixotroph whose photoautotrophic range is extended by heterotrophy. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

A rapid determination of histamine and 3-methylhistamine in human plasma by high performance liquid chromatography

Histamine plays an important pathophysiological role in allergy, inflammation, gastric acid secretion, microcirculation and neurotransmission. 3-Methylhistamine is a prominent metabolite of histamine. Different methods for determination of histamine in biological fluids have been developed. In the present study, a simple, simultaneous determination of histamine and 3-methylhistamine by HPLC (precolumn derivatization with fluorescamine) was developed in human plasma using fluorescence detection with 1-methylhistamine as the internal standard. Linear regression analysis of the ratios of the concentrations of histamine and 3-methylhistamine (X) against peak height ratios (Y) yielded the following: y = 0.0073x ? 0.0096 (R ² = 0.990) and y = 0.0077x ? 0.0111 (R ² = 0.989). In conclusion, it was possible to detect histamine and 3-methylhistamine below 5 ng mL^?1 in 1 mL plasma.