Determination of Total Flavonoids in Acanthopanax senticosus(Rupr. et Maxim.) Harms by Enzymolysis and Its Technological Optimization

In order to optimize the ultrasonic extraction technique for the total flavonoid of leaf yellows plus, the contents of 21 leaf yellows plus total flavonoid from four regions in Heilongjiang Province were comparatively analyzed. The ultrasonic extraction technology was optimized by Box-Behnken response surface method, and the total flavonoid content of 21 kinds of Acanthopanax senticosus(Rupr. et Maxim.) Harms from different producing areas were analyzed by cluster analysis. The optimal process conditions were determined as ultrasonic time 30 min, solid-liquid ratio 1 : 12 and ultrasonic power 250 W, and the average yield of the total flavonoid was 1.453 mg·g^ (-1). By optimizing the ultrasonic-assisted extraction method, the total flavonoid content from different producing areas was compared in the experiment, which provided certain data support for the optimization of the extraction process in the future and laid a certain theoretical foundation for the quality analysis of Chinese medicinal materials.

Use of a rat model to characterize 35 arterial pulse wave parameters in a comparative study of isoflurane and Zoletil/ xylazine anesthesia and the effect of Acanthopanax senticosus extract

Background : Information obtained from arterial pulse waveforms (APW) can be usefulfor characterizing the cardiovascular system. To achieve this, it is necessary to know thedetailed characteristics of APWs in different states of an organism, which would allowAPW parameters (APW- Ps) to be assigned to particular (patho)physiological conditions.Therefore, our work aimed to characterize 35 APW- Ps in rats under the influence ofisoflurane (ISO) and Zoletil/xylazine (ZO/XY) anesthesia and to study the effect of rootextract from Acanthopanax senticosus (ASRE) in these anesthetic conditions.Methods : The right jugular vein of anesthetized rats was cannulated for the administrationof ASRE and the left carotid artery for the detection of APWs from which 35APW- Ps were evaluated.Results : We obtained data on 35 APW- Ps, which significantly depended on the anesthesia,and thus, they characterized the cardiovascular system under these two conditions.ASRE transiently modulated all 35 APW- Ps, including a transient decrease insystolic and diastolic blood pressure (BP) and heart rate or increases in pulse BP, d P /d t max , and systolic and diastolic areas. Whereas the transient effects of ASRE weresimilar, the extract had prolonged disturbing effects on the cardiovascular system inrats under ZO/XY but not under ISO anesthesia. This negative effect can result fromthe disturbance caused by ZO/XY anesthesia on the cardiovascular system.Conclusions : We characterized 35 APW- Ps of rats under ISO and ZO/XY anesthesiaand found that ASRE contains compounds that can modulate the properties of thecardiovascular system, which significantly depended on the status of the cardiovascularsystem. This should be considered when using ASRE as a nutritional supplementby individuals with cardiovascular problems.

Determination of Eleutheroside B and E in Acanthopanax Preparations by High-P erformance Liquid Chromatography with Solid-Phase Extraction

Aim An HPLC method for analyzing eleutheroside B (ELU B) and eleutheroside E(ELU E) , two of the main active substances of Acanthopanax preparations were studied. Methods Thesamples were analyzed on a kromasil ODS column with water-acetonitrile as a gradient mobile phase.The flow rate was 0.8 mL·min^(-1) and detecting wavelengths were 206 nm for ELU B, 220 nm for ELUE, solid phase extraction (SPE) and internal standard-salicin were selected. Results The recoveriesof Acanthopanax tablets and injection were 90.4% - 96.8% and 96.4% - 99.8% for ELU B, 87.7% -93.3%and 95.7% - 98.5% for ELU E, respectively. The linear ranges were 4.45 - 22.25 μg· mL^(-1) (r =0.999 8) and 5.11 - 25.55 μg·mL^(-1) ( r = 0.999 7) respectively. Conclusion This method can savethe time for cleaning the chromatographic system and improve sensitivity for Acanthopanaxpreparations , thus providing a way to evaluate the quality of Acanthopanax preparations.

短柄五加(Acanthopanax brachypus)rbcL基因的结构分析

克隆了含完整短柄五加rbcL基因的3.2kb EcoRI片段,测定了该基因的核苷酸序列.所测核苷酸序列总长度为1924bp,其中编码区1428bp,编码475个氨基酸的蛋白质.测定的基因5’上游区共278bp,包含原核性质-35区(TTGCGC),-10区(TACAAT)及类似真核的TATA box元件(TATATA).5’前导区长194bp,其中SD序列为GGAGG,紧邻起始密码子上游.测定的3’下游区共218bp,含2个相邻的转录后可形成茎环结构的反向重复序列.短柄五加rbcL基因编码区推导的氨基酸序列与烟草、菠菜、豌豆、苜蓿、玉米、水稻、松树、地钱、衣藻和Anacystis的同源性分别为93.5％、94.11％、94.53％、94.74％、89.68％、92.21％、92.21％、92.63％、87.58％和80.84％.本文还对不同植物rbcL基因的启动区及部分5’和3’非编码区进行了比较分析.

白簕(Acanthopanax trifoliatus (L.) Merr)种子的休眠机理研究

药蔬两用的白簕(Acanthopanax trifoliatus(L.)Merr)深受人们喜爱并广泛栽培,种子具有一定的休眠特性.为了探索白簕种子的休眠机理,本研究解剖种子的内部结构、测定种壳(内果皮)的透水性和透气性以及提取果实和种子各部分內源抑制物质并进行生物测定.研究结果表明:1)种胚细小,约为0.3-0.5mm,位于种子一角隅,可能存在生理后熟.2)种壳存在一定的机械束缚和透水透气障碍,相对剥皮种子和缺刻种子,可能导致完整种子萌发启动时间延后.3)白簕果实和种子各部分都可能含有抑制物质,其中果皮中存在活性较强的內源抑制物质,抑制活性为外果皮及果肉>内果皮>完整果实>完整种子>种胚和胚乳,且白簕外果皮及果肉、内果皮及完整果实随着抑制物浓度的增加,抑制作用显著增强.由此推测,白簕种子属于生理休眠类型.本研究初步阐明白簕种子的休眠机理,以期为其有性繁殖和打破休眠提供理论依据.

超声波法提取刺五加(Acanthopanax senticosus)中丁香甙的研究

通过均匀设计的试验方法,探讨了用超声波法提取刺五加根茎中丁香甙的工艺条件,确定的最佳工艺条件为:水为溶剂,溶剂用量8ml/g,超声提取200min,提取温度为室温。

辽东栎林下和开阔地上短柄五加(Acanthopanax brachpus)无性系种群生长的比较研究

研究了分布于开阔地和辽东栎林下短柄五加无性系种群的扩散方式、无性系的形态可塑性和分株的死亡年龄。研究结果表明 ,开阔地上无性系分株的扩散方式为稀疏线型 ,根茎长度、无性系分株数目 ,林下和开阔地差异显著。根茎分枝角度、无性系根茎总长无显著性差异。

大孔吸附树脂提取刺五加(Acanthopanax senticosus)丁香苷

Adsorptive resin was used to extract syringin from Acanthopanax senticosus, and the content of syringin was assayed by HPLC. Some factors affecting the extraction efficiency such as resin type, resin amount, eluent type, eluent flow rate and amount were examined. The optimal purification condition was selected.

Acanthopanax senticosus polysaccharides-induced intestinal tight junction injury alleviation via inhibition of NF-κB/MLCK pathway in a mouse endotoxemia model

AIM To examine the effects of Acanthopanax senticosus polysaccharides(ASPS) on intestinal tight junction(TJ) disruption and nuclear factor-kappa B(NF-κB)/myosin light chain kinase(MLCK) activation in endotoxemia.METHODS BALB/C mice(6-8-weeks-old) received continuous intragastric gavage of ASPS for 7 d before injection of lipopolysaccharide(LPS), or received ASPS once after LPS injection. Blood and intestinal mucosal samples were collected 6 h after LPS challenge. Clinical symptoms, histological injury, intestinal permeability,TJ ultrastructure, and TJ protein expression were determined.RESULTS Compared with mice in the LPS group, pretreatment with ASPS improved clinical and histological scores by 390.9%(P < 0.05) and 57.89%(P < 0.05), respectively, and gut permeability change in endotoxemic mice was shown by a 61.93% reduction in reduced leakage of fluorescein isothiocyanate-dextran 6 h after LPS injection(P < 0.05). ASPS pretreatment also prevented LPS-induced TJ ultrastructure breakdown supported by increased electron dense materials between adjoining cells, sustained redistribution and expression of occludin(0.597 ± 0.027 vs 0.103 ± 0.009, P < 0.05) and zonula occludens-1(0.507 ± 0.032 vs 0.125 ± 0.019, P < 0.05), and suppressed activation of the NF-κB/MLCK pathway indicated by reduced expression of NF-κB, phospho-inhibitor kappa B-alpha, MLCK and phospho-myosin light-chain-2 by 16.06%(P < 0.05), 54.31%(P < 0.05), 66.10%(P < 0.05) and 64.82%(P < 0.05), respectively. CONCLUSION ASPS pretreatment may be associated with inhibition of the NF-κB/MLCK pathway and concomitant amelioration of LPS-induced TJ dysfunction of intestinal epithelium in endotoxemia.

Effects of Acanthopanax senticosus on learning and memory in a mouse model of Alzheimer's disease and protection against free radical injury to brain tissue

BACKGROUND： Acanthopanax senticosus, a plant of the Araliaceae family, is used in traditional Chinese medicine. It can be used to replenish Qi, strengthen the spleen, tonify the kidney, and relieve mental strain. OBJECTIVE： To observe effects ofAcanthopanax senticosus on learning and memory in a mouse model of Alzheimer＇s disease and abnormal biochemical changes in the brain tissue. DESIGN： A completely randomized grouping, controlled animal experiment. SETTING： Department of Preventive Medicine, School of Basic Medical Sciences, Yanbian University. MATERIALS： A total of 50 Kunming mice, aged 1-1.5 months, equal numbers of males and females, were provided by the Laboratory Animal Center, Yanbian University Medical College. The study was performed in accordance with ethical guidelines for the use and care of animals. Acanthopanax was provided by Yanbian Chengda Pharmaceutical Co., Ltd. Acanthopanax senticosus （0.5 kg） was soaked in water for 1 hour and transferred to 1.5 kg distilled water for extraction. It was boiled for 1 hour and extracted after 1 hour of boiling. The procedure was repeated 3 times. The extract was condensed to 500 mL and then adjusted to 500 and 1000 g/L with water. Piracetam tablets were produced by Shandong Luoxin Pharmaceutical Corporation, China. Malonaldehyde （MDA）, superoxide dismutase （SOD）, and acetylcholinesterase （ACHE） kits were purchased from Nanjing Jiancheng Bioengineering Co., Ltd., China. METHODS： This study was performed at the Department of Preventive Medicine, School of Basic Medical Sciences, Yanbian University from January to June 2007. All mice were randomly divided into 5 groups with 10 mice in each： control group, model group, low-, and high-dose Acanthopanax senticosus-treated groups, and piracetam-treated group. All groups were administered 0.1 mL/10 g. In the control and model groups, mice were intragastrically administered saline each morning for 5 days prior to experimentation. Five days later, they were intraperitoneally perfused with saline and aluminum trichloride （100 mg/kg）, respectively, every other afternoon. In the low- and high-dose Acanthopanax senticosus-treated groups, as well as the piracetam-treated groups, mice were intragastrically administered 500 and 1 000 g/L Acanthopanax senticosus and 23 g/L piracetam suspension, respectively, every morning. Five days later, they were intraperitoneally perfused with aluminum trichloride （100 mg/kg） every other afternoon, 20 successive times. MAIN OUTCOME MEASURES： On days 20, 23, 26, and 29 after treatment, time-to-platform, number of errors, and accuracy were measured by water maze. After anesthesia, mice were euthanized and whole brain tissues were immediately resected and homogenized. MDA levels, and SOD and AChE activities, were measured using the corresponding kits. RESULTS： Fifty mice were included in the final analysis. In the model group, accuracy increased, and time-to-platform was prolonged, compared to control group （P 〈 0.05 0.01）. In the model group, MDA levels significantly increased （P 〈 0.05）, while SOD activity significantly decreased （P 〈 0.01）, compared to control group. MDA levels were significantly lower and SOD activity was significantly higher in the low- and high-dose Acanthopanax senticosus-treated groups, compared to the model group （P 〈 0.05 0.01）. In the model group, AChE activity significantly increased, compared to the control group （P 〈 0.05）, AChE activity was significantly lower in the high-dose Acanthopanax senticosus-treated and the piracetam-treated groups than in the model group （P 〈 0.05）. CONCLUSION： Acanthopanax senticosus can improve learning and memory in a mouse model of Alzheimer＇s disease, and concomitantly increase SOD activity, inhibit AChE activity, and decrease MDA levels.

RAPD analysis of natural populations of Acanthopanax brachypus

Random Amplified Polymorphic DNA (RAPD) analysis is a new technology of molecular marking which has proved very powerful in detecting genetic diversity at the level of population. The genomic DNAs used in our experiment were extracted from fresh leaves taken from 59 individuals sampled from three natural populations in Yan An, Shanxi Province. Through more than 2,000 PCRs, deep-going RAPD analysis was carried out on DNA samples from 49 inviduals. The percentage of polymorphic RAPD loci found in these three populations were respectively 27.2 %, 18.6 % and 5.4 %; the average genetic distances within population, 0.055, 0.036 and 0.008; the average genetic distances between populations (Ⅰ-Ⅱ), (Ⅰ-Ⅲ) and (Ⅱ-Ⅲ), 0.105, 0.096 and 0.060. The genetic diversity of A. brachypus within and between populations was found, for the first time, to be rather poor,thus revealing innate factors as the cause contributing to its endangered status. In addition, our work also provides basic materials for elucidating the underlying cause of its endangerment and for its protection biology.

Acanthopanax senticosus aqueous extract protects PC12 cells against hypoxic injury

Acanthopanax senticosus aqueous extract, at 10, 5 and 2.5 μg/mL, lessened hypoxia-induced PC12 cell injury. Pretreatment with the extract upregulated Bcl-2 expression, downregulated Bax expression, elevated superoxide dismutase activity, diminished malondialdehyde content, relieved Ca2＋ overloading, and suppressed apoptosis of PC12 cells. These effects were most pronounced at 10 μg/mL. These results suggest that Acanthopanax senticosus aqueous extract protects PC12 cells from hypoxia-induced injury in a dose-dependent manner.

Three new compounds from the leaves of Acanthopanax senticosus Harms

Three new compounds named (2E)prenyl benzoate-4-O-α-L-arabinopyranosyl (1→6) β-D-glucopyranoside (1), 7-methoxy-8-O-β-D-glucopyranosyl coumarin (2), and 3,4'-dihydroxy-3'-methoxy benzenepentanoic acid (3) were isolated from the leaves of Acanthopanax senticosus Harms. The structures of new compounds were determined by means of 2D NMR experiments and chemical methods.

Advances in Research of Acanthopanax trifoliatus( L. ) Merr

Acanthopanax trifoliatus( L.) Merr is a common traditional Chinese herbal medicine in central and southern regions of China. It has high medicinal and edible value and mainly contains flavonoids,polysaccharides,saponins and volatile components. Its active extracts have antioxidant,anti-inflammatory,anti-fatigue,and analgesic effects. From the biological characteristics,effective chemical components,extraction methods,cultivation techniques,pharmacological effects,toxicological activities,and product development status,this paper reviewed in detail the research status of A. trifoliatus in China and abroad,to provide a scientific basis for in-depth research and development,and utilization of A. trifoliatus.

Characterization of stereostructure by X-ray and technology of extracting in combination hydrolysis in situ of acankoreanogenin from leaves of Acanthopanax gracilistylus W.W.Smith

To study the stereostructure by X-ray and the technology of extracting acankoreanogenin from the leaves of Acanthopanax graeilistylus W. W. Smith （AGS）, the crystal structure was measured with a Bruker APEX-Ⅱ area-detector diffractometer instrument and the technology of extracting in combination hydrolysis in situ （ECHS） was compared with these of traditional methods. The crystal belongs to the monoclinic system, space group P2b with unit cell parameters： a=（8.3652±0.0006） nm, b=（24.721±0.002） nm, and c=（14.5587±0.0011） nm, α=90°, β=97.850 （4） °, γ=90 °, V=2982.51 nm3, Dc= 1.179 mg/m3, and the molecular number （Z） of elementary structures was 2. The comparisons show that the extraction rate of acankoreanogenin with ECHS methods is much higher than that of traditional methods. Then, central composite design-response surface methodology （CCD-RSM） was adopted for optimizing the extraction rate of ECHS methods. The optimized values of extraction parameters are as follows： for the for extraction process of acid hydrolysis are that extraction time 110.8 min, solvent-herb ratio 11.5 and acid content 5.25%; the best extraction process of basic hydrolysis are that extract time 120 min, solvent-herb ratio 8.7 and the alkali content 8.79%. Finally, the extracts were purified with decolorizing carbon after alkali solution and acid-isolation and purity of acankoreanogenin was 98.7%.

Microwave Assisted-extraction of Flavonols from Acanthopanax Senticosus Harms and High-performance Liquid Chromatography Analysis

Flavonols were extracted from Acanthopanax Senticosus Harms by Soxhlet extraction（SE）,atmospheric pressure microwave-assisted extraction（AMAE） and pressurized microwave-assisted extraction（PMAE）.The influences of experimental conditions,such as concentration of ethanol in the eluent,microwave irradiation time,etc.on the extraction yield were studied.Four flavonols（hyperin,rutin,quercitrin and quercetin） in the extracts were determined quantitatively by HPLC method.The experimental results demonstrate the advantages of PMAE and AMAE over those of conventional SE,that is the extraction time is dramatically reduced,and the yields of flavonols are effective improved.Although the yield of flavonols is higher and the extraction time is shorter by PMAE than those of AMAE,flavonols are easy to be decomposed over a certain time.

THE INFLUENCE OF HETEROGENEOUS HABITATS ON THE GERMINATION OF ACANTHOPANAX SENTICOSUS SEEDS AND THE DYNAMICS OF ITS SEED POOL

Differences in germination of .A. Senticosus, an important shrub under the Korean Pine broad-leavd forest and its secondary forest in heterogeneous habitats and the dynamics of its seed pool have been studied by observations in location. The results showed that transformation rates from seeds to seedlings were in the following order : 16.8% in Larch plantations, 4. 1% in Mongolian oak forests, 2.7% in Birch forests, 1.8% in Korean Pine plantations, and 0.5% in hard wood forests.The life-span of seeds was determined to be four years in the simulating seed pool under Larch plantations. The percentage of the seedling output was, 14.5% in the 2nd year, 10. 1% in the 3rd year, and 1 .8% in the 4th year. Other types of the seed output were those f decayed 33. 1 %, senescent, 22.3%,predated by mice 14. 1%, which was variable due to the variation in the number of mouse, and only 1. 17% were eaten by insects and other soil animals.

刺五加(Acanthopanax senticosus)生理生态学特性的研究——1.展叶期

本文研究了刺五加展叶期的生理生态学特性。结果表明,林冠层下面光强度较弱(荫蔽度为41—71％),相对湿度高、温度变化小、创造一个特殊的森林小气候,形成一个较为阴湿的环境。在这样的生境条件下,刺五加的蒸腾强度在0.407—1.816g·g^(-1)FW·h^(-1)之间,平均值为0.943g·g^(-1)FW·h^(-1),光合强度为48.607mgDW·dm^(-2)·h^(-1),其中净光合强度为39.305mgDW·dm^(-2)·h^(-1),呼吸强度为9.302mgDW·dm^(-2)·h^(-1)。叶绿素含量达4.565mg·g^(-1)DW叶绿素a/b值为0.4。

五加属(Acanthopanax)五加组植物的数量分类研究

以五加属五加组（Ｓｅｃｔ．Ａｃａｎｔｈｏｐａｎａｘ）植物１１个种为分类单位，以２２个形态特征作为性状，用系统聚类的方法探索了种与种之间的亲缘关系，认为浙江五加Ａｃａｎｔｈｏｐａｎａｘｚｈｅ－ｊｉａｎｇｅｎｓｅＸ．Ｊ．Ｘｕｅ与细刺五加Ａ．ｓｅｔｕｌｏｓｕｓＦｒａｎｃｈ在形态上有明显差异。

北京地区刺五加(Acanthopanax senticosus)群落优势种种间关联性分析

为了探讨北京地区刺五加(Acanthopanax senticosus)群落的物种组成、群落结构和种间关联性,在典型抽样法调查的基础上,基于2×2列联表,采用方差比率法、χ2检验、Pearson相关系数和Spearman秩相关系数检验等数量分析方法,对北京百花山和雾灵山自然保护区刺五加群落中的27个优势物种、351个种对间的种间关联性进行了研究。结果显示,刺五加群落中27个优势物种总体呈极显著正关联,表明该群落处于相对稳定的演替阶段;χ2检验表明,正关联种对有191对,负关联种对有160对,正、负关联种对比值为1.19;Pearson相关检验发现,正关联的种对有112对,负关联的种对有239对,正、负关联种对比值为0.47;Spearman秩相关检验结果显示,正关联的种对有170对,负关联的种对有181对,正、负关联种对比值为0.94。将优势物种对环境的适应方式和主导生态因素相结合进行PCA排序发现,27个优势物种可被划分为4个生态种组。本研究结果表明北京地区刺五加群落物种组成相对稳定且群落结构正处于发展阶段。