五味子治疗大鼠糖尿病肾病作用机制的血清代谢组学研究

采用基于超高效液相色谱与串联四级杆飞行时间质谱仪(Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry,UPLC/Q-TOF-MS)联用技术的代谢组学方法,通过分析大鼠血清内源性代谢物的变化,研究五味子治疗糖尿病肾病的作用机制。利用高脂高糖饲料喂养并腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型。给药12周后,采用试剂盒方法测定尿蛋白、尿肌酐的含量,结果表明五味子水提取物可以显著降低模型动物的尿蛋白含量(p<0.05),对糖尿病大鼠肾病并发症具有一定的改善作用。采用UPLC/Q-TOF-MS方法分析了五味子对糖尿病肾病大鼠的血清代谢轮廓,分析了健康组、模型组和五味子给药组的大鼠血清,采用偏最小二乘法-判别分析(Partial least squares discriminant analysis,PLS-DA)进行数据分析。PLS-DA得分图显示健康组、模型组和五味子组的代谢轮廓有显著差别,根据正交偏最小二乘法-判别分析(Orthogonal partial least squares discriminant analysis,OPLS-DA)载荷图筛选,将对各组分离贡献大的化合物的串联质谱分析数据,经Human Metabolome Database(HMDB)等数据库检索,进行质谱信息匹配,鉴定出黄尿酸、油酰胺、棕榈酰胺、尿酸、5-羟基己酸、硫酸对甲酚、对甲酚葡萄糖苷酸7种内源性代谢物为生物标记物。研究结果表明五味子通过影响色氨酸代谢、嘌呤代谢、肠内菌代谢、脂肪酸代谢等通路对糖尿病肾病发挥治疗作用,其中嘌呤代谢、肠内菌代谢通路可能是五味子发挥治疗作用的重要途径。     

Separation and purification of 9,10‐dihydrophenanthrenes and bibenzyls from Pholidota chinensis by high‐speed countercurrent chromatography

Stilbenoids are the main components of leaves and stems of Pholidota chinensis. In the present investigation, high‐speed counter‐current chromatography was used for the separation and purification of two classes of stilbenoids, namely, bibenzyls and 9,10‐dihydrophenanthrenes, on a preparative scale from whole plants of P. chinensis with different solvent systems after silica gel column chromatography fractionation. n‐Hexane/ethyl acetate/methanol/water (1.2:1:1:0.8, v/v/v/v) was selected as the optimum solvent system to purify 1‐(3,4,5‐trimethoxyphenyl)‐1′,2′‐ethanediol ( 1 ), coelonin ( 2 ), 3,4′‐dihydroxy‐5,5′‐dimethoxybibenzyl ( 3 ), and 2,?7‐?dihydroxy‐?3,?4,?6‐?trimethoxy‐?9,?10‐?dihydrophenanthrene ( 4 ). While 2,7‐dihydroxy‐3,4,6‐trimethoxy‐?9,?10‐?dihydrophenanthrene ( 5 ), batatasin III ( 6 ), orchinol ( 7 ), and 3′‐O‐methylbatatasin III ( 8 ) were purified by n‐hexane/ethyl acetate/methanol/water (1.6:0.8:1.2:0.4, v/v/v/v). After the high‐speed counter‐current chromatography isolation procedure, the purity of all compounds was over 94% assayed by ultra high performance liquid chromatography. The chemical structure identification of all compounds was carried out by mass spectrometry and ¹H and ¹³C NMR spectroscopy. To the best of our knowledge, the current investigation is the first study for the separation and purification of bibenzyls and 9,10‐dihydrophenanthrenes by high‐speed counter‐current chromatography from natural resources.     

Selective and sensitive determination of protoberberines by capillary electrophoresis coupled with molecularly imprinted microextraction

In this work, we developed a novel molecularly imprinted solid‐phase microextraction with capillary electrophoresis method for the selective extraction and determination of protoberberines in complicated samples. The imprinted monolith was prepared in a micropipette tip‐based device by using acrylamide as the functional monomer, ethyleneglyoldimethacrylate as the cross‐linker and dimethylsulfoxide as the porogen, and exhibited an imprinting factor of 2.41 to berberine, 2.36 to palmatine and 2.38 to jatrorrhizine. Good capillary electrophoresis separation was achieved by using 20 mM phosphate buffer at pH 7 as running buffer with the addition of organic modifier of 10% methanol. Parameters such as sample pH value, sample flow rate and sample volume were investigated for imprinted monolith‐based solid‐phase microextraction. An imprinted solid‐phase microextraction with capillary electrophoresis method was developed, the method showed a wide linear range (0.3–50 μg/mL), good linearity (R² ≥ 0.9947) and good reproducibility (relative standard deviations ≤ 0.73%), the limit of detection was as low as 0.1 μg/mL, which was lower than some reported methods based on capillary electrophoresis for protoberberines. The method has been applied for determination of three common protoberberines in Cortex Phellodendri Chinensis, by using a molecularly imprinted monolith as the selective sorbent, most of the matrices in the Cortex Phellodendri Chinensis sample were removed and three protoberberines were selectively enriched and well determined.     

Anti-diabetic Effects of Polysaccharides from Ethanol-insoluble Residue of Schisandra chinensis(Turcz.) Baill on Alloxan-induced Diabetic Mice

The ethanol-insoluble residue of Schisandra generated during lignans industrial production is usually treated as solid waste. However, there is active polysaccharide which could be used in it. In this work, the water-soluble polysaccharides from the ethanol-insoluble residue of Schisandra(ESCP) were obtained and their anti-diabetic effect was evaluated. The results indicate that ESCP could significantly reduce the blood glucose level in alloxan-induced diabetic mice. Moreover, the ESCP could significantly improve the lipid metabolism and increase the content of liver glycogen in alloxan-induced diabetic mice. The results indicate that ESCP could be developed into a potential natural hypoglycemic agent.     

透明质酸酶的色氨酸残基修饰与荧光光谱研究

北五味子[Schisandra chinensis（Turcz．）Baill．]属广义木兰科植物,主产于我国东北,故又称“辽五味”,中药五味子的主要药材为北五味子的干燥果实,作为一种传统中药,五味子具有收敛固涩,益气生津,补肾宁心的功效,用于肺喘虚咳,心悸失眠诸病。     

基于TG-FTIR的圆柏心、边材热解研究

生物质热解是实现生物质废弃物有效处理及生物质材料高效利用的重要途径之一,因此对生物质热解过程及其机理进行研究具有极大的现实意义。木材作为一种来源广泛的可再生材料也是众多生物质热解原料中的一种,由于组成成分等多方面的差别,不同树种的热解特性具有明显差异。然而,同一种木材的心材与边材由于在组织构造、化学组成及其含量等方面有着明显不同,因此在热解特性和产物方面也存在一定的差异,该工作即对此进行深入研究。试验以心、边材区分明显的常见园林绿化物圆柏为试验材料,通过TG-FTIR联用技术分别得到其心、边材热解过程中的热重曲线和挥发分的红外光谱谱图并对其进行分析。结果表明,纤维素、半纤维素、木质素和抽提物含量对心、边材热解特性影响显著。由于圆柏心材半纤维素、木质素含量较高,在热解前期和热解后期具有比边材更大的失重率,而较高的抽提物含量则在一定程度上增加了其在反应中期的失重率,减小了该范围内心、边材失重率的差异;边材纤维素含量较高,因此其在300~380 ℃,失重明显,在DTG曲线中的最大失重速率也较高。在红外图谱中,心、边材热解所产生的挥发分种类几乎相同,但在数量上有差异,当热解达到最大失重速率时,所产生的挥发分明显增加,且整个过程中边材产生的有机酸类化合物更多,而心材相对产生更多的水和CO₂。     

Two new noroleanane-type triterpenoid saponins from the stems of Stauntonia chinensis

Two new noroleanane-type triterpenoid saponins, 3β,20α,24-trihydroxy-29-norolean-12-en-28-oic acid 24-O-β-L-fucopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-β-D-glucopyranoside (1) and 3β,20α,24-trihydroxy-29-norolean-12-en-28-oic acid 24-O-β-D-glucopyranosyl-(1→2)-[α-L-arabinopyranosyl-(1→3)]-β-D-glucopyranoside (2) were isolated from the stems of Stauntonia chinensis DC., together with three known compounds, brachyantheraoside B₂ (3), eupteleasaponin Ⅷ (4) and fargoside B (5). Their structures were elucidated by spectroscopic and chemical methods. The cytotoxic activities of compounds 1 and 2 were evaluated against five human tumor cell lines (HCT-116, HepG2, BGC-823, NCI-H1650, and A2780). Compounds 1 and 2 showed moderate cytotoxic activities toward the tested cell lines with IC₅₀ values ranging from 12.71 to 32.04 μM.     

Application of characteristic fragment filtering with ultra high performance liquid chromatography coupled with high‐resolution mass spectrometry for comprehensive identification of components in Schisandrae chinensis Fructus

An integrated strategy of characteristic fragment filtering combined with target database screening based on ultra‐high‐performance liquid chromatography coupled with high‐resolution mass spectrometry was proposed for comprehensive profiling of components in Schisandrae chinensis Fructus. The strategy consisted of following five steps: (1) Representative standards were analyzed by ultra high performance liquid chromatography coupled with linear ion trap‐Orbitrap mass spectrometer for characteristic fragments and fragmentation rules of each structure type. (2) The raw data of 70% methanol extract was collected by ultra high performance liquid chromatography quadrupole time‐of‐flight tandem mass spectrometry. (3) The chemical components database that consisted of names, chemical formulas and structures of potential components in Schisandrae chinensis Fructus was established by summarizing previous literature to screen the collected liquid chromatography with mass spectrometry data and obtain matched compounds. (4) Characteristic fragments, literature, and reference standards were used to verify the matches. (5) Characteristic fragment filtering combined with online database querying was used to deduce potential new compounds. As a result, a total of 94 compounds were identified or characterized and 16 of them were potential new compounds. The study provided a reference for comprehensive characterization of ingredients in herbal medicine and formed the foundation for pharmacodynamic study of Schisandrae chinensis Fructus.     

开口箭化学成分及抗肿瘤活性

从开口箭新鲜根茎中分离得到8个化合物, 通过理化性质分析、 质谱、 红外光谱及核磁共振波谱检测等方法鉴定了其结构, 分别为开口箭皂苷J(1)、 (25S)-26-O-β-D-吡喃葡萄糖基呋甾-1β,3β,22α,26-四羟基-3-O-β-D-吡喃葡萄糖苷(2)、 洋地黄毒苷元-3-O-α-L-吡喃岩藻糖苷(3)、 弯蕊皂苷元B(4)、 异罗斯考皂苷元(5)、 罗斯考皂苷元(6)、 香豌豆酚(7)及(+)-儿茶素(8). 其中, 化合物1为新化合物, 化合物3, 4, 7和8为首次从该属植物中分离得到. 在抗人结肠癌细胞LoVo和胃癌细胞BGC-823活性评价中, 化合物5和6表现出较强的抗肿瘤活性, 半抑制浓度(IC₅₀)值分别达到0.532和0.757 μmol/L.     

Seconoriridone A: A C16-seco-noriridal derivative with a 5/5/7 tricyclic skeleton from Belamcanda chinensis

Seconoriridone A (1), a novel C₁₆-seco-noriridal with a 5/5/7 tricyclic scaffold, was isolated from the rhizome of Belamcanda chinensis. Its structure and absolute configuration were determined by NMR spectroscopy and quantum chemical calculations. Notably, seconoriridone A featured a 2-butanonyl moiety with oxidative cleavage of the conventionally multisubstituted cyclohexane ring of iridals. Seconoriridone A was a mixture of C-14 epimers in the presence of hemiacetal. The plausible biosynthetic pathway for 1 was deduced.