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.

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。

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.

Determination of A canthopanax senticosus ＇ s Seed Viability

以6个产地的刺五加种子为试验材料，采用四唑染色法（TTC法）、红墨水染色法、溴麝香草酚蓝法（BTB法）、炒种法及纸上荧光法测定种子生活力，筛选最佳的刺五加种子生活力测定方法。结果显示：溴麝香草酚蓝法是测定刺五加种子生活力的最佳方法。

Cloning and Sequence Analysis of ATPase Beta Subunit Gene from Eleutherococcus senticosus

[Objective] This study aimed to clone and analyze the cDNA of ATPase β subunit gene from Eleutherococcus senticosus.[Method] A pair of homologous primers was designed according to the chloroplast ATPase β subunit gene sequences of the known species;then the gene cDNA of E.senticosus were amplified by RT-PCR and compared with that of the known species;its structure was predicted finally.[Result] 1 099 bp of ATPase beta subunit cDNA of E.senticosus which encodes 366 amino acids was amplified by RT-PCR.Sequence comparison and structure prediction showed that amino acids encoded by the ATPase beta subunit gene of E.senticosus shared the highest homology,up to 96.41% with that of Oryza sativa.In the secondary structure,the protein contained 171 alpha helixes accounting for 46.72%,53 extended strands accounting for 14.48%,27 beta sheets accounting for 7.38% and 115 random coils which took up 31.42%.The amino acids 262-271 were the symbolic site of ATPase β subunit.The whole peptide chain had no obvious hydrophobic region and was primarily confirmed as a hydrophilic protein.[Conclusion] The cNDA of ATPase β subunit gene cloned from E.senticosus in this study will provide reference for learning the effect of energy metabolism on secondary metabolism,structure and function of ATPase in E.senticosus.

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。

Plasmolysis treatment enhances the expression of callose synthase gene in zygotic embryos of Eleutherococcus senticosus

In previous study we reported that pretreatment with plasmolysis enhanced somatic embryo formation in hypocotyls of Eleutherococcus senticosus.In the present study,the expression level of callose synthase gene in embryos of E.senticosus in response to 2,4-D,sucrose and mannitol treatments was analyzed by RT-PCR.The results show that plasmolysis pretreatment using sucrose and mannitol significantly promoted the expression of callose synthase gene.Also,the thicker cell walls of explant plasmolyzed compared with controls were observed during the somatic embryogenesis.We suggest that the callose may make the cells in epidermis separate from neighboring cells and then develop into embryogenic potential cells.

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。

Supercritical Fluid Extraction of Total Flavonoids from Leaves of Acanthopanax Senticosus Harms

The herbal plant Acanthopanax Senticosus Harms is natural herb of Changbaishan in Jilin Province of China, which belongs to the Araliaceae family. As the ingredients of folk medicine, it has long been used to treat a variety of human diseases, such as cerebrovascular disease, diabetes, tumor, isochemic heart diseases, hypertension, rheumatic arthritis, etc. E2,33. Flavonoids, a class of constituent compounds, which have a broad distribution in the nature and are found in Acanthopanax Senticosus Harms, have gained particular attention. A number of studies have shown that flavonoid compounds have wide biological activities, such as anti-aging, anticancer, anti-HIV,

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.

Developing conservation strategies for Pinus koraiensis and Eleutherococcus senticosus by using model-based geographic distributions

We described potential changes in the geo- graphic distribution and occurrence probability of Pinus koraiensis Sieb. et Zucc. and Eleutherococcus senticosus （Rupr. et Maxim.） Maxim. in the counties of northeast China. This information was used to identify priority areas for protection and provide protection and management recommendations within each studied county. The two species were mapped in 2884 study plots throughout this region over a 4-year period （38°40＇N-53°30＇N, 115°05＇E- 135°02＇E）. We used the species distribution models （Maxent）, systematic conservation planning models （Marxan）, and Geographic Information Systems （ArcGIS 10.0）. The distributions of two species were correlated in the study area, enabling unique and economically viable joint conservation measures to be implemented. Three models were combined to identify feasible priority con- servation sites. We used local spatial statistics to assess all identified conservation areas in relation to potential climate change based shifts in the geographic distribution of the two species. Model-based conservation strategies were used to identify effective measures to protect and utilize these two tree species in the study region. This study pre- sents a novel technique for assessing wild plant distribu- tions, in addition to serving as a model for the conservation of other species within the framework of general forest management, ecological construction, and geographical surveying.

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.

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.

<i>Eleutherococcus senticosus</i>: Studies and effects

Ginseng is one of the most popular herbal supplements in the world. It is a plant widely used in folk and traditional medicines for cardiovascular, immune, nervous and endocrine systems, and according to the researchers, it has the ability to increase the non-specific resistance state, which characterizes it as an adaptogenic substance. There are different species of ginseng, such as the American, Chinese, Korean and Japanese ginseng;the Korean species (Panax ginseng) is being used for thousands of years as a tonic, prophylactic and “restorative” agent, with powerful antioxidant properties. For a long time, its use was empirical, because people used to believe that it was a panacea that promoted longevity, with beneficial effects for the treatment of physical fatigues. Nowadays, the active components of Eleutherococcus senticosus are well described, however, there are no data on the quantity of a certain class of these secondary compounds produced in each species. Although the Eleutherococcus senticosus extract may contain several substances, including vitamins, minerals, cellulose, and ethanol, the substances responsible for inducing various physiological responses are the eleutherosides (in the root) and ciwujianosides (in the leaf). As Eleutherococcus senticosus receives great attention by showing that its active components can provide protection against oxidative stress, among other benefits, contributing to health and the prevention and treatment of diseases, such as diabetes, cancer, cardiovascular disease and inflammation. The purpose of this article is to describe the main, adverse and toxicological effects of Eleutherococcus senticosus recently related in the literature.

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.

The Effect of Methylation Modification of MDD on the Expression of Key Genes in the Process of Saponin Synthesis in Eleutherococcus senticosus

Mevalonate pyrophosphate decarboxylase is a kind of key enzyme in the terpenoid synthesis pathway in Eleutherococcus senticosus.The results of bisulfite sequencing showed that there were three kinds of samples with a low(0.68%),medium(0.72%)and high(0.79%)DNA methylation ratio in the promoter of MDD in E.senticosus,respectively.The transcriptome sequencing results showed that the expression of MDD in E.senticosus was significantly up-regulated in the types with low DNA methylation ratios of MDD(P<0.05).There was a significant negative correlation between the saponin content in E.senticosus and the DNA methylation ratio of MDD promoter(P<0.05).The screening results of differentially expressed transcription factors among the three groups with different DNA methylation ratio showed that a total of 4 transcription factors could bind to 6 CpG sites.Protein-protein interaction analysis showed that E.senticosus MDD could interacted with other key enzymes in the process of terpenoid synthesis.In addition,it was found that the DNA methylation of MDD promoter was mainly regulated by DNA methyltransferase.These results demonstrated that under the action of DNA methyltransferase,the changes of DNA methylation of MDD promoter could regulate its own expression level by affecting the combination ability of transcription factors,and then affected the expression of other genes in terpenoid synthesis pathway.The synthesis and accumulation of saponins in E.senticosus was also changed.

刺五加多酚合成纳米氧化锌及其抗氧化和抗菌性能研究

本试验旨在绿色合成纳米氧化锌并探究该纳米氧化锌的活性。以刺五加多酚为原料,绿色合成纳米氧化锌(ZnO NPs),通过紫外-可见分光光度计、X-射线衍射仪(XRD)、傅里叶变换红外光谱仪(FTIR)、扫描电镜(SEM)、透射电镜(TEM)、热重分析(TG)和粒径分析对所制得的样品进行表征,通过1,1-二苯基-2-三硝基苯肼(DPPH)和2,2′-联氮-双-3-乙基苯并噻唑啉-6-磺酸(ABTS)自由基清除试验评价其抗氧化性能,通过沙门氏菌、大肠杆菌和金黄色葡萄球菌抑菌试验评价其抗菌性能。结果表明:本试验成功合成ZnO NPs,其在361 nm处有吸收峰,形态为六方晶系纤锌矿结构,晶体平均粒径为77.45 nm;当ZnO NPs质量浓度达到6 g/L时,对DPPH自由基的清除率达到93.28%;当ZnO NPs质量浓度达到9 g/L时,对ABTS自由基的清除率达到73.83%;ZnO NPs对沙门氏菌、大肠杆菌的最小抑菌浓度为64 g/L,最小杀菌浓度为80 g/L,对金黄色葡萄球菌的最小抑菌浓度为80 g/L,最小杀菌浓度为88 g/L,且ZnO NPs对大肠杆菌、沙门氏菌、金黄色葡萄球菌的抑菌活性均高于氧化锌(ZnO)。本试验以刺五加多酚为原料成功合成ZnO NPs,该ZnO NPs具有良好的抗氧化和抗菌性能,为其进一步开发为饲料添加剂奠定了基础。