A novel flavonoid from the root of Macrothelypteris torresiana(Gaud.) Ching

A novel flavonoid was isolated from the root of Macrothelypteris torresiana （Gaud.） Ching. The structure of the product was identified as 5, 7-dihydroxy-2-（1, 2-isopropyldioxy-4-oxocyclohex-5-enyl）-chromen-4-one on the ground of chemical and spectro-scopic methods.2009 Jin Lan Ruan. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Two new flavanone glycosides from Macrothelypteris torresiana (Gaud.) Ching

Two new flavanone glycosides 1 and 2 were isolated from the aerial parts of Macrothelypteris torresiana （Gaud.） Ching. The structures of two products were identified as （2S）-5,7,2＇,5＇-tetrahydroxyflavanone-2＇-O-β-D-6＂-O-acetylglucopyranoside and （2S）- 5,7,2＇,5＇-tetrahydroxyflavanone-2＇-O-β-D-glucopyranoside on the basis of their chemical and spectral analysis, respectively.

Preparation of Magnetic Molecularly Imprinted Polymers for Selective Isolation and Determination of Kaempferol and Protoapigenone in Macrothelypteris torresiana

Novel uniform-sized magnetic molecularly imprinted polymers （MMIPs） were synthe- sized for selective recognition of active antitumor ingredients of kaempferol （KMF） and protoapi- genone （PA） in Macrothelypteris torresiana （M. torresiana） by surface molecular imprinting tech- nique in this study. Super paramagnetic core-sheU nanoparticles （γ-MPS-SiO2@Fe3O4） were used as seeds, KMF as template molecule, acrylamide （AM） as functional monomer, and N, N＇-methylene bisacrylamide （BisAM） as cross-linker. The prepared MMIPs were characterized by X-ray diffraction （XRD）, Fourier transform infrared spectrum fiT/R）, transmission electron microscopy （TEM） and thermo-gravimetric analysis （TGA）, respectively. The recognition capacity of MMIPs was 2.436 times of non-imprinted polymers. The adsorption results based on kinetics and isotherm analysis were in accordance with the pseudo-second-order model （R2=0.9980） and the Langmuir adsorption model （R2=0.9944）. The value of E （6.742 kJ/mol） calculated from the Dubinin-Radushkevich isotherm model suggested that the physical adsorption via hydrogen-bonding might be predominant. The Scatchard plot showed a single line （R2=0.9172） and demonstrated the homogeneous recognition sites on MMIPs for KMF. The magnetic solid phase extraction （MSPE） based on MMIPs as sorbent was established for fast and selective enrichment of KMF and its structural analogue PA from the crude extract of M. torresiana and then KMF and PA were detected by HPLC-UV. The established method showed good performance and satisfactory results for real sample analysis. It also showed the feasi- bility of MMIPs for selective recognition of active structural analogues from complex herbal extracts.

Hepatoprotective activity of Macrothelypteris torresiana (Gaudich.) aerial parts against CCl4-induced hepatotoxicity in rodents and analysis of polyphenolic compounds by HPTLC

Macrothelypteris torresiana is a fern species belonging to the family Thelypteridaceae. The present study was conducted to evaluate hepatoprotective potential of ethanol extract from M. torresiana aerial parts(EEMTAP)and detect the polyphenolic compounds present in the extract using high performance thin layer chromatography(HPTLC). Hepatoprotective potential of EEMTAP were tested at doses of 300 and 600 mg/kg, per os(p.o.), on Wistar albino rats. The extract and silymarin treated animal groups showed significant decrease in activities of different biochemical parameters like serum glutamic oxaloacetic transaminase(SGOT), serum glutamate-pyruvate transaminase(SGPT), and alkaline phosphatase(ALP), which were elevated by carbon tetrachloride(CCl_4) intoxication. The levels of total bilirubin and total protein along with the liver weight were also restored to normalcy by EEMTAP and silymarin treatment. After CCl_4 administration, the levels of hepatic antioxidant enzymes such as glutathione(GSH) and catalase(CAT) were decreased whereas the level of hepatic lipid peroxidation(LPO) was elevated. The levels of these hepatic antioxidant enzymes were also brought to normalcy by EEMTAP and silymarin treatment. Histological studies supported the biochemical findings, and treatment with EEMTAP at doses of 300 and 600 mg/kg, p.o. was found to be effective in restoring CCl_4-induced hepatotoxicity in rats. A simple HPTLC analysis was conducted for the detection of polyphenolic compounds in EEMTAP, and the result revealed the presence of caffeic acid as phenolic acid and quercetin as flavonoid. The proposed HPTLC method is simple and concise and provides a good resolution of caffeic acid and quercetin from other constituents present in EEMTAP.

普通针毛蕨配子体发育的研究

采用混合土培养普通针毛蕨（Macrothelypteris torresiana （Gaud．） Ching）孢子，显微镜下观察记录其孢子萌发及配子体发育过程。结果表明：孢子褐色，赤道面观为半圆形，极面观椭圆形，单裂缝。播种3d左右孢子萌发，萌发类型为书带蕨型，配子体发育为槲蕨型。播种7d左右发育为片状体，丝状体与片状体两阶段的界限不明显。原叶体边缘及背腹面均可产生毛状体。播种21d左右发育形成幼原叶体，成熟原叶体呈心脏形。播种35d左右开始有性器官出现，精子器近圆球形，由3个细胞组成，成熟颈卵器颈部由4列细胞组成，4～5层细胞高。原叶体受精后1个月内即可观察到从原叶体上生成的幼胚。

普通针毛蕨质量标准的研究

目的研究普通针毛蕨的质量标准。方法运用显微鉴定、薄层色谱、紫外可见分光光度法、高效液相色谱等技术对普通针毛蕨进行研究,对其水分、总灰分、酸不溶灰分、总黄酮、原芹菜素含量进行测定。结果参照《中国药典》2005年版(Ⅰ部),普通针毛蕨生药中水分不得超过15.00%,总灰分不得超过8.31%,酸不溶性灰分不得超过3.90%,t检验显示,上述指标,因产地不同,无显著性差异(P>0.05);生药中原芹菜素的含量不得低于0.71%,总黄酮含量不得低于28.21%。因药材产地不同,其总黄酮、原芹菜素含量有显著性差异(0.01<P≤0.05)。结论实验结果为建立普通针毛蕨药材的质量标准提供了依据,方法简单易行。

Inhibition of Ageratina adenophora on Spore Germination and Gametophyte Development of Macrothelypteris torresiana

Allelopathy of Ageratina adenophora plays an important role in its invasion. However, we have little knowledge of its allelpathic effects on ferns. In Petri dish bioassays, the inhibitory potential of aqueous leachates from roots, stems and leaves of A. adenophora was studied on the spore germination and gametophyte development of Macrothelypteris torresiana. All leachatss inhibited the spore germination and growth of the first rhizoid of M. torresiana and inhibitory effects increased with increasing leachate concentrations. Root leachates proved most inhibitory. Gametophyte rhizoids of M. torresiana treated with stem and leaf leachates of A. adenophora were erect, which was similar to those of the control. However, gametophyte rhizoids of M. torresiana treated with root leachates of A. adenophora were erect, but also curving or swollen. Moreover, curving and swollen rhizoids increased with increasing concentrations. As time went by, rhizoids treated with root leachates were not so curved and the swelling almost disappeared. Possible causes are discussed in the present study. The increasing concentrations of leaf leachates also delayed the stages of gametophyte development. With the treatment of root leachates, the delay was more obvious. Thus A. adenophora inhibited the spore germination and gametophyte development of M. torresiana and the root leachates were most inhibitory.

正交试验法优选普通针毛蕨地上部位总黄酮提取纯化工艺

目的:优选普通针毛蕨地上部位总黄酮提取和纯化工艺条件。方法:以总黄酮提取率为考查指标,液料比、提取次数、提取时间、乙醇浓度为考察因素,选用L9(34)表对提取工艺进行正交试验。以总黄酮提取率、总黄酮纯度和主要活性成分原芹菜素(protoapigenone)物质为考查指标,用高效液相色谱法测定原芹菜素含量,以加碱水量,加碱水次数,碱水pH值,加酸后pH为影响因素,选用L9(34)表对纯化工艺进行正交试验。结果:普通针毛蕨总黄酮提取最佳工艺为20倍量75%乙醇提取3次,每次1.5h。纯化条件为药材醇提浸膏加2倍量pH9的氢氧化钠溶液,提取2次,提取液用浓盐酸调节为pH1。结论:优选工艺获得的总黄酮提取率,纯度和原芹菜素含量较高。本方法合理,便于生产。

普通针毛蕨颈卵器和卵的发育

蕨类植物卵发生是有性生殖研究的重要内容。真水龙骨类II的卵发生尚无人研究,本文对该类群中的普通针毛蕨(Macrothelypteris torresiana)卵发生过程进行了光镜和透射电镜研究。结果显示:卵细胞刚形成时,与腹沟细胞紧密相连,随着卵进一步发育,卵细胞和腹沟细胞之间逐渐形成分离腔,但孔区处卵细胞和腹沟细胞始终相连。随后,卵细胞上表面有不定型物质堆积在质膜外,形成一层加厚的卵膜,孔区处没有卵膜覆盖的位置最后形成受精孔。在卵发育后期,卵细胞核变得不规则,近成熟时卵核产生大量核外突。卵发育过程中卵膜的出现、受精孔的产生以及核外突等特征,与进化的真水龙骨类中其他蕨类植物的卵发生研究结果相似,与薄囊蕨类中原始基部类群的卵发生现象有显著差别。由此可见,普通针毛蕨属于进化类型。依据卵发生中卵膜和受精孔等特征推测原始薄囊蕨类经过里白类、桫椤类,最终演化为水龙骨类。

普通针毛蕨总黄酮包合物的制备及其生物利用度研究

目的:了解普通针毛蕨总黄酮及其包合物的活性成分在人工胃、肠液中的稳定性及其生物利用度。方法:以人工胃、肠液将普通针毛蕨总黄酮0.5%CMC-Na混悬剂及HP-β-CD包合物制成高、中、低3个浓度样品,在温孵箱内37℃恒温、振荡50r.min-1,于0、4、8、12、20、24h取样品进行HPLC检测,检测其活性成分代表原芹菜素的峰面积进行比较;高、中、低3种相等剂量(按体质量分别以100,200,300mg.kg-1给药)的普通针毛蕨总黄酮混悬剂及其包合物进行动物灌胃血液吸收动力学实验,采用Das 2.0软件对各组血药浓度进行房室模型分析。结果:普通针毛蕨总黄酮及其包合物的所有样品在0～24h内,同一样品间所含的原芹菜素峰面积差异无显著性(P>0.05),相同浓度的样品组与阳性组、阴性组比较所含的原芹菜素峰面积无显著性差异(P>0.05),说明原芹菜素在胃肠液中比较稳定,不易破坏;普通针毛蕨总黄酮混悬剂及其包合物的大鼠灌胃血液吸收动力学试验,表现出明显的线性动力学特征,显示包合物延长了半衰期、血药浓度达峰时间变短,峰浓度有所提高,消除半衰期相对延长,AUC相对加大,说明包合处理可提高总黄酮口服吸收的生物利用度。结论:普通针毛蕨总黄酮中的活性成分在人工胃肠液稳定性强,不易破坏;包合物的生物利用度显著提高。

普通针毛蕨中原芹菜素与其糖苷对PDK-1/AKT途径的拮抗效应

目的:揭示原芹菜素与其糖苷之间的拮抗作用,并在分子水平上阐明这两个化合物对于PDK-1介导的AKT磷酸化的拮抗作用。方法:首先建立普通针毛蕨的HPLC指纹图谱,并确定目标化合物含量。而后通过免疫印迹实验,采用组合模式给药,证实了这两个化合物对于PDK-1介导的AKT磷酸化的拮抗作用。再后通过CETSA实验进一步证实了这两个化合物对PDK-1蛋白的竞争性结合行为,最后采用分子对接给予这种拮抗作用图像化的解释。结果:普通针毛蕨提取物中原芹菜素和原芹菜素葡萄糖苷的摩尔浓度分布为1.249μmol·L^-1和0.928μmol·L^-1。在该浓度下原芹菜素的抗肿瘤增殖效果可以被原芹菜素葡萄糖苷所抑制,且原芹菜素所介导的PDK-1下调也可以被原芹菜素葡萄糖苷所拮抗。同时CETSA实验进一步证实了这个两个化合物与PDK-1的竞争性结合。结论:本研究揭示了一个原芹菜素的拮抗因素,并为原芹菜素葡萄糖苷含量控制提供了依据。