刺五加种子结构,后熟作用及其细胞化学研究

刺五加种子为扁肾形,种皮由一层细胞构成。种子脱落时,胚处于心形胚期,胚周围的胚乳细胞解体形成囊腔包囊胚,胚细胞原生质浓厚,胚乳细胞中贮存大量蛋白质和脂类,但两者均未见贮存多糖,有萌发潜能的种子只占全部种子的１２．８０％,种子经变温层积处理６个月即可完成后熟过程,其细胞化学特点是：处理１．５个月时胚细胞中开始积累多糖颗粒,至４个月时达最大量并一直保持至种子萌发。试验地种植条件下饱满种子经１８－２０个     

刺五加胚和胚乳发育的研究

刺五加Eleutherococcus senticosus(Rupr .et Maxim．)Maxim的胚胎发生类型为茄型。其卵细胞受精后,合子经历15天左右的休眠期才进行第一次分裂。合子分裂通常发生在胚乳细胞化之后,经棒形胚、球形胚、至果实成熟时发育到心形胚。棒形胚后期至心形胚初期,胚柄最为发达。刺五加的胚乳发育类型为核型。其初生胚乳核的休眠期为1天左右。当胚乳游离核数目增加到200至300时,胚乳以自由生长细胞壁的方式细胞化,胚乳细胞以典型的有丝分裂方式进一步增殖,增加细胞数目。球形胚时期,胚乳细胞内开始贮藏营养物质。少数种子的胚乳里存在巨大细胞核的异型胚乳细胞。在胚乳游离核为32至64个时,分化出珠被绒毡层;球形胚时期,珠被绒毡层解体。珠被绒毡层解体后,胚乳表层细胞分化为分泌层。球形胚至心形胚阶段,约有5％的种子里,胚与胚乳组织发生弥散样降解。成熟果实中,含有大量的瘪粒种子和虫咬种子;饱满种子率为40％左右。饱满种子中,胚乳组织占据种子体积的绝大部分,胚所占比率很小。讨论了不同发育时期胚和胚乳的营养供应。     

刺五加、短梗五加的开花动态及繁育系统的比较研究

野外定位观测刺五加（Eleutherococcus senticosus),短梗五回（E.sessiliflorus)的开花进程,花朵的功能形态特征和开花动态,用杂交指数（OCI）,花粉－胚珠比（P／O）,去雄,套袋,人工授粉等方法分别测定刺五加,短梗五加的繁育系统,结果显示,刺五加种群,短梗五加种群的花期均持续1个月左右,刺五加比短梗五加早开花20d 左右,二均有雄蕊先熟现象,刺五加是单全异株植物,种群内既具有雄性,又具有雌株,还具有两性株,繁育系统主要为异交,需要传粉活动才能完成授粉过程,与刺五加不同,短梗五加仅具两性花,但繁育系统也以异交为主,短梗五加两性花中的雌,雄器官既在空间上分离,又在时间上分离,只能进行同株异花间或异株,异花间传粉才能受精结实。     

Protective Effects of 3,4-Seco-Lupane Triterpenes from Food Raw Materials of the Leaves of Eleutherococcus Senticosus and Eleutherococcus Sessiliflorus on Arrhythmia Induced by Barium Chloride

As a traditional wild vegetable and food raw material, the leaves of Eleutherococcus senticosus and Eleutherococcus sessiliflorus are rich in 3,4-seco-lupane triterpenes, including chiisanoside (CSS), divaroside (DVS), sessiloside-A (SSA), and chiisanogenin (CSG). This study was conducted to evaluate the anti-arrhythmic effects of these 3,4-seco-lupane triterpenes. Evaluation of the cytotoxicity of compounds was performed by measuring cell viability and apoptosis with the CCK-8 assay. In vivo, arrhythmia was induced by rapid injection of BaCl₂ via rat caudal vein. The occurrence time and duration of arrhythmias in rats were studied. The levels of SOD and MDA in serum, and Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase in myocardial homogenate were detected by ELISA. The histopathological changes of rats myocardial were observed by HE staining. Changes in the expression of PKA and related proteins were detected by Western blot. The 3,4-seco-lupane triterpenes interactions with protein kinase A were analyzed by molecular docking. In the present study, we found that 3,4-seco-lupane triterpenes exhibited powerful anti-arrhythmic activity, especially DVS completely relieved the ventricular arrhythmia induced by BaCl₂. This study suggests that the leaves of E. senticosus and E. sessiliflorus might be used as functional food materials to prevent arrhythmia, and DVS can potentially be further developed as an anti-arrhythmic drug.     

Determination and pharmacokinetic study of chiisanogenin in rat plasma by ultra performance liquid chromatography-tandem mass spectrometry

Introduction – Chiisanogenin existing in many Acanthopanax species has been reported to possess anti‐inflammatory, antibacterial and antiplatelet aggregatory activities. Objective – To develop and validate a rapid and sensitive ultra performance liquid chromatography‐tandem mass spectrometry method for the determination of chiisanogenin in rat plasma and to investigate its pharmacokinetics after oral administration of chiisanogenin or the extract of Acanthopanax sessiliflorus fruits. Methodology – The sample pretreatment involved a one‐step extraction of 0.2 mL plasma with diethyl ether. Acetaminophen was used as the internal standard. The separation was carried out on an ACQUITY UPLC? BEH C₁₈ column with a mobile phase of acetonitrile‐5 mM ammonium acetate (90:10, v/v) at a flow rate of 0.2 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI) source. Results – A high sample throughput was achieved with an analysis time of 1.1 min per sample. The calibration curve was linear (r² ≥ 0.99) over the concentration range of 5–500 ng/mL with a lower limit of quantification (LLOQ) of 5 ng/mL. The intra‐day and inter‐day precision (relative standard deviation, R.S.D.) values were below 11% and the accuracy (relative error, R.E.) was within 8% at all three quality control (QC) levels. Conclusion – The method was successfully applied to the pharmacokinetic study of chiisanogenin in rat after oral administration of chiisanogenin and the extract of Acanthopanax sessiliflorus fruits. Other constituents in the extract affected the pharmacokinetic behavior of chiisanogenin. Copyright © 2010 John Wiley & Sons, Ltd.     

无梗五加根中苯丙素类化合物的研究

从无梗五加(Acanthoparax sessiliflorus( Rupr.et Maxim.)Seem.)根70％乙醇提取物的乙酸乙酯层中提取分离得到8个苯丙素类化合物.经理化和波谱分析鉴定为(+)-表芝麻脂素(1)、(-)-芝麻脂素(2)、赛菊芋黄素(3)、洒维宁(4)、咖啡酸甲酯(5)、对羟基桂皮酸(6)、(-)-丁香脂素(7)、(+)-松脂索(8).化合物8是首次从五加科植物中分离得到,化合物1和5是首次从五加属植物中分离得到,化合物3、6和7是首次从该植物中分离得到.     

刺五加鲨烯合酶基因cDNA的克隆与序列分析

采用改良的异硫氰酸胍法提取刺五加总RNA,逆转录为cDNA,根据已报道的人参鲨烯合酶基因(squalene synthase gene,SS) cDNA序列设计引物,利用RT-PCR法克隆刺五加SS基因的cDNA序列.克隆得到长度为1258 bp的刺五加SS基因cDNA序列,开放阅读框全长1248 bp,编码415个氨基酸残基,GenBank登录号为HQ456918,与人参的SS1、SS2和SS3氨基酸序列一致性分别为91.73％、97.59％和96.63％.首次分离并报道了刺五加SS基因cDNA序列,为刺五加苷生物合成中关键酶的表达分析及调控机理研究提供参考.     

Impact of Chisan (ADAPT-232) on the quality-of-life and its efficacy as an adjuvant in the treatment of acute non-specific pneumonia

A double-blind, placebo-controlled, randomized (simple randomisation), pilot (phase III) study of Chisan^® (ADAPT-232; a standardised fixed combination of extracts of Rhodiola rosea L., Schisandra chinensis Turcz. Baill., and Eleutherococcus senticosus Maxim) was carried out on two parallel groups of patients suffering from acute non-specific pneumonia. Sixty patients (males and females; 18–65 years old) received a standard treatment with cephazoline, bromhexine, and theophylline: in addition, one group of 30 patients was given Chisan mixture, whilst the second group of 30 patients received a placebo, each medication being taken twice daily from the beginning of the study for 10–15 days. The primary outcome measurements were the duration of antibiotic therapy associated with the clinical manifestations of the acute phase of the disease, together with an evaluation of mental performance in a psychometric test and the self-evaluation of quality-of-life (QOL) (WHOQOL-Bref questionnaires) before treatment and on the first and fifth days after clinical convalescence. The mean duration of treatment with antibiotics required to bring about recovery from the acute phase of the disease was 2 days shorter in patients treated with Chisan compared with those in the placebo group. With respect to all QOL domains (physical, psychological, social and ecological), patients in the Chisan group scored higher at the beginning of the rehabilitation period, and significantly higher on the fifth day after clinical convalescence, than patients in the control group. Clearly, adjuvant therapy with ADAPT-232 has a positive effect on the recovery of patients by decreasing the duration of the acute phase of the illness, by increasing mental performance of patients in the rehabilitation period, and by improving their QOL. Both the clinical and laboratory results of the present study suggest that Chisan (ADAPT-232) can be recommended in the standard treatment of patients with acute non-specific pneumonia as an adjuvant to increase the QOL and to expedite the recovery of patients.     

Quality control of roots of Eleutherococcus senticosus by HPLC

An HPLC method based on several known methods for the determination of eleutherosides B and E was developed, optimised and validated in terms of linearity, precision (repeatability and intermediate precision on different days and at different concentration levels) and accuracy (recovery). The extraction procedure, the extraction solvent and the extraction yield were evaluated and optimised. A reversed-phase RP-18 column gradient eluted with a two-phase system consisting of phosphoric acid:water (0.5:99.5) and acetonitrile was used to evaluate the samples; detection was at 220 nm. Although eleutherosides B and E are commercially available, they are very costly, and therefore ferulic acid was chosen as external standard. The correction factors for the response of ferulic acid against both eleutherosides were determined and validated. This method, accepted by the European Pharmacopoeia Commission, will be included in the monograph on Eleutherococcus senticosus roots to assay the content of eleutherosides B and E.