高效液相色谱-高分辨质谱法鉴定拉萨大黄化学成分

建立了高效液相色谱-高分辨质谱法分析鉴定拉萨大黄化学成分。选用Agilent Zorbax SB-C18柱（150 mm×4.6 mm×5 μm）,以乙腈和水为流动相进行梯度洗脱,流速0.5 mL/min;采用电喷雾离子源,在正、负离子模式下对拉萨大黄提取物进行一级和多级全扫描质谱分析。综合分析化合物的色谱和质谱行为,并结合对照品、文献和数据库等相关数据鉴定化合物。在负离子模式下,鉴定出63种化合物;正离子模式下,鉴定出54种化合物。所检测到的化合物包括22种鞣质类、28种茋类、6种黄酮类、3种苯丙酸类、2种苯丁酮类和2种有机酸类,其中有53种化合物为在该药材中首次发现,且未发现蒽醌类成分。该方法快速、灵敏,不仅提高了拉萨大黄复杂基质中微量化合物的鉴定效率,同时实现了对拉萨大黄提取物中不同类型化合物的分析鉴别,为进一步阐明拉萨大黄的药效物质基础及质控标准提供了科学依据。     

Hepatoprotective Potency of Chrysophanol 8-O-Glucoside from Rheum palmatum L. against Hepatic Fibrosis via Regulation of the STAT3 Signaling Pathway

Rhubarb is a well-known herb worldwide and includes approximately 60 species of the Rheum genus. One of the representative plants is Rheum palmatum, which is prescribed as official rhubarb due to its pharmacological potential in the Korean and Chinese pharmacopoeia. In our bioactive screening, we found out that the EtOH extract of R. palmatum inhibited hepatic stellate cell (HSC) activation by transforming growth factor β1 (TGF-β1). Chemical investigation of the EtOH extract led to the isolation of chrysophanol 8-O-glucoside, which was determined by structural analysis using NMR spectroscopic techniques and electrospray ionization mass spectrometry (ESIMS). To elucidate the effects of chrysophanol 8-O-glucoside on HSC activation, activated LX-2 cells were treated for 48 h with chrysophanol 8-O-glucoside, and α-SMA and collagen, HSC activation markers, were measured by comparative quantitative real-time PCR (qPCR) and western blotting analysis. Chrysophanol 8-O-glucoside significantly inhibited the protein and mRNA expression of α-SMA and collagen compared with that in TGF-β1-treated LX-2 cells. Next, the expression of phosphorylated SMAD2 (p-SMAD2) and p-STAT3 was measured and the translocation of p-STAT3 to the nucleus was analyzed by western blotting analysis. The expression of p-SMAD2 and p-STAT3 showed that chrysophanol 8-O-glucoside strongly downregulated STAT3 phosphorylation by inhibiting the nuclear translocation of p-STAT3, which is an important mechanism in HSC activation. Moreover, chrysophanol 8-O-glucoside suppressed the expression of p-p38, not that of p-JNK or p-Erk, which can activate STAT3 phosphorylation and inhibit MMP2 expression, the downstream target of STAT3 signaling. These findings provided experimental evidence concerning the hepatoprotective effects of chrysophanol 8-O-glucoside against liver damage and revealed the molecular basis underlying its anti-fibrotic effects through the blocking of HSC activation.     

Screening of plant species for the phytotreatment of wastewater containing sulphonated anthraquinones

Sulphonated anthraquinones are known to be recalcitrant to biodegradation and are not eliminated by traditional wastewater treatment plants, leading to their accumulation in fresh water. Due to the high cost and limited efficiency of existing physical-chemical treatments, alternative cheaper processes are required to remove these compounds from industrial effluents. Four plant species were tested under hydroponic conditions for their ability to treat model effluents contaminated with mono- and disulphonated anthraquinones. Among them, Rheum rabarbarum (rhubarb) showed the most promising results and was chosen for further investigation. The apparent transpiration stream concentration factor obtained with this plant species reached up to 2.5, indicating a strong phytotreatment potential that should be further explored then exploited.     

DPPH法评价大黄提取物的抗氧化活性

采用超声波法提取大黄抗氧化物质,以DPPH法评价大黄抗氧化活性。结果表明,大黄抗氧化物质最佳提取工艺为:以70%乙醇为溶剂,料液比为1∶40 g/mL,超声提取2次,每次提取30 min;大黄抗氧化活性物质主要分布于石油醚和乙酸乙酯提取物,为该药材的开发提供参考。     

唐古特大黄多糖对大鼠应激性胃溃疡的保护作用

目的: 探讨唐古特大黄多糖(Rheum tanguticum polysaccharides, RTP) 对大鼠应激性胃溃疡的保护作用。方法: 用水浸束缚应激法(water immersion and restraint stress, WRS) 复制大鼠应激性胃溃疡模型, 提前灌胃给予大黄多糖, 应激6 h 后处死动物,观察溃疡指数和胃粘膜损伤程度的变化, 检测血清及胃粘膜组织中超氧物歧化酶(SOD) 和丙二醛(MDA) 的变化。结果: RTP 能明显降低应激大鼠胃粘膜溃疡指数和胃粘膜MDA 水平, 升高血清和胃粘膜SOD 活性。结论: RTP 对水浸束缚应激引起的大鼠应激性胃溃疡有明显的保护作用, 可能是大黄治疗应激性胃溃疡的有效成分之一。     

Identification of the antidiarrhoeal components in official rhubarb using liquid chromatography–tandem mass spectrometry

Though the purgative components in official rhubarb are well documented as anthraquinone glycosides and sennosides, the antidiarrhoeal effect and its chemical basis have not been reported before. In this study, liquid chromatography coupled with electrospray ionisation tandem mass spectrometry was used to investigate the main chemicals in the antidiarrhoeal fraction previously isolated from Rheum palmatum L. In total 10 compounds were identified in the fraction as follows: gallic acid, galloyl glucose, di-O-galloyl-glucose, glucopyranosyl-galloyl-glucose, coumaroyl-O-galloyl-glucose, trimer of catechin, catechin gallate, catechin-glucopyranoside, carboxyl-chrysophanol-O-glucose and emodin-O-glucose. The predominant components identified in the fraction were rhubarb tannins, except for two anthraquinone glycosides of low contents. The polyphenolic analysis also showed the occurrence of abundant tannins. So, the antidiarrhoeal basis of rhubarb was primarily proposed as tannin-related compounds.     

Phytoconstituents and therapeutic uses of Rheum emodi wall. ex Meissn

Rhubarb (Rheum emodi, family Polygonaceae) has been traditionally used as diuretic, liver stimulant, purgative/cathartic, stomachic, anticholesterolaemic, antitumour, antiseptic and tonic. A number of anthraquinone derivatives including emodin, aloe-emodin, physcion, chrysophanol, rhein, emodin glycoside and chrysophanol glycoside occur as the main chemical constituents. In the past few years, new components such as sulfemodin 8-O-β-d-glucoside, revandchinone-1, revandchinone-2, revandchinone-3, revandchinone-4, 6-methyl-rhein and 6-methyl aloe-emodin have been reported from the same species. Anthraquinone derivatives show evidence of antifungal, anti-microbial, anti-Parkinson’s, anti-proliferative, immuno-enhancing, antiviral and antioxidant activities. This review covers published work on botany, chemistry and therapeutic uses of different components from rhubarb.     

Immuno-enhancing effects of the herbal extract from Himalayan rhubarb Rheum emodi Wall. ex Meissn

In the present studies, the immuno-enhancing activity of Rheum emodi ethyl acetate rhizome extract has been carried out by conducting studies on the cell lines. The studies include effects on signal transduction parameters that could pave the way for understanding some of the treatment principles of the plant extracts. The data indicate that R. emodi may have an immuno-enhancing effect through the release of various cytokines. There is a dose dependent increase in the release of NO (nitric oxide) and cytokine TNF-α, IL-12 and a decrease in IL-10 by RAW 264.7 in macrophage cell lines in the presence of extract alone. However the cytokine level is concomitantly increased in the presence of stimulant as well as extract. The up-regulation of TNF-α, and IL-12 up to the level of 200 ng/ml and 530 ng/ml respectively switches the cell towards Th-1 immune response. TNF-α, IL-12 induces generation and proliferation of Th-1 cells which in turn produces NK and cytotoxic T-cells. On the other hand, the decreased levels of IL-10 switches off the Th-2 immune system and directs the cell towards Th-1 immune response only. Thus, indicating that R. emodi extract may have anti-tumour effects by regulating Th-1 and Th-2 immune response in vivo.     

黄棒菜超氧化物歧化酶(SOD)分离纯化的技术

以植物新品种黄棒菜为原料,研究料液比,浸提温度及pH,热变性温度及时间、丙酮加入量等因素对黄棒菜超氧化物歧化酶(SOD)提取效果的影响,利用正交试验与单因素试验优化提取工艺。结果表明,提取SOD的最佳条件为:料液比1︰3(g/mL),浸提温度0℃,pH 7.0,50℃热变性20 min,1.5倍酶液体积的丙酮,此时SOD纯化倍数可达3.17。经层析纯化及真空冷冻干燥,获得SOD成品。最佳条件下处理新鲜黄棒菜叶茎、叶片组织,测得SOD最终比活力分别为7662.23 U/mg,4897.39 U/mg,纯化倍数达17.55、23.08,叶茎更适于提取SOD。经电泳检测,黄棒菜SOD相对分子质量约80 kDa,据文献鉴定为Mn-SOD。试验结果为黄棒菜SOD的分离纯化及资源开发提供技术参考。