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.

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.

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.

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.

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.

Acanthopanax senticosus Protects Structure and Function of Mesencephalic Mitochondria in A Mouse Model of Parkinson's Disease

Objective： To investigate the neuro-protective effects of Acanthopanax senticosus Harms（EAS） on mesencephalic mitochondria and the mechanism of action, using a mouse model of Parkinson＇s disease（PD）. Methods： The chemical fingerprint analysis of the extract of Acanthopanax senticosus Harms（EAS） was performed using the ultra performance liquid chromatograph and time of flight mass spectrometry. Thirty mice were randomly divided into the control group, the MPTP model group, and the EAS treated group with MPTP（MPTP＋EAS group, 10 in each group）. The MPTP model group and the MPTP＋EAS group received MPTP-HCl（30 mg/kg i.p） once a day for 5 days. The control group received an equal volume of saline（20 m L/kg i.p） once a day for 5 days. Induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride daily（MPTP-HCl, 30 mg/kg） for 5 days, the PD mice were treated with EAS at 45.5 mg/kg daily for 20 days. The behavioral testing of mice was carried out using the pole-climbing test. The integrity and functions of neurons were examined in mesencephalic mitochondria in a PD mouse model, including nicotinamide adenine dinucleotide dehydrogenase ubiquinone flavoprotein 2（NDUFV2）, mitochondrially encoded nicotinamide adenine dinucleotide dehydrogenase 1（MT-ND1）, succinate dehydrogenase complex subunit A（SDHA）, and succinate dehydrogenase cytochrome b560 subunit（SDHC）. Results： After treatment with EAS, the behavioral changes induced by MPTP were attenuated significantly（P〈0.05）. EAS protected the mesencephalic mitochondria from swelling and attenuated the decreases in their membrane potential（both P〈0.05）, which was supported by an ultra-structural level analysis. The changes in reactive oxygen species（ROS）, malonic dialdehyde（MDA）, oxidative phosphorylation（OXPHOS） system 4 subunits levels and PD-related proteins expressions（parkin, Pink1, DJ-1, α-synuclein, and Lrrk2） reverted to near normal levels（all P〈0.05）, based on the results of immune-histological and Western blotting observations. Conclusions： The neuro-protective effects of EAS are linked to protecting mice against MPTPinduced mitochondrial dysfunction and structuraldamage.Therefore,EAS is a promising candidate for the prevention or treatment of mitochondrial neurodegenerative disorders,such as PD.

Protection of Acanthopanax Senticosus Saponin on Free Radical Injury Induced Aging of Nerve Cell

Objective To study the effect of Acanthopanax senticosus saponin (ASS) on free radical injury induced neuron aging. Methods On day 7 of fetal mice, cortical neuron primary passage cultures were divided into the normal control group, model group and ASS groups. The model group using free radical (FeSO4 plus H2O2) injury mode prepared in vivo cultured ICR mice cortical neuron aging model; ASS groups: 24 hrs before and after treated with H2O2 and FeSO4, different concentration of ASS was added, according to biochemical parameters such as lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) etc. and histomorphologic change to observe the protection of ASS on aging neurons. Results The LDH, SOD,MDA of the model group were compared with the normal group, < 0. 01; ASS groups added 1.25 mg/100 ml, 2.5 mg/100 ml, 5 mg/100 ml concentration of ASS, their LDH, SOD, MDA compared with the model group π.05-0.01, the difference was significant. In medicated groups the SOD activity of oxidization injured nerve cells obviously elevated, LDH activity and MDA content apparently lowered. Microscope and scanning electron microscopic observation showed that supplemented with ASS to protect the nerve cell injury abated, part of the cellular structure tended to normalize. Conclusion ASS could act against free radical toxic effect, increase the anti-oxidase activity, strengthen the protection of neuron cells. It is assumed that the effect against nerve cell aging was possibly through scavenging oxygen free radical, strengthening the stability of cell membrane, thus delaying the development of aging.

Extrusion process of Acanthopanax senticosus leaves enhances the gastroprotective effect of compound 48/80 on acute gastric mucosal lesion in rats

OBJECTIVE:To investigate the gastroprotective effects of Acanthopanax senticosus leaves(ASLs)extrusion on acute gastric mucosal lesion in rats induced by compound 48/80(C48/80).METHODS:Rats were divided into six groups:normal;C48/80-induced gastric lesion control;gastric lesion positive control(famotidine 4 mg/kg);gastric lesion administered with two levels of extruded ASLs(ASLE,40 and 200 mg/kg);and gastric lesion treated with ASLs(ASL 200 mg/kg).Mucus secretion/damage was determined by immunohistological staining.Immunofluorescence and western blotting were performed to determine gastric mucosal Bax and Bcl-2 expression.Gastric mucosal oxidative-stress-related enzymes and malondialdehyde were determined.RESULTS:C48/80-induced mucus depletion and inflammation in the gastric mucosa were significantly attenuated by ASLs.The increased serum serotonin and histamine concentrations in C48/80-treated rats were also attenuated by ASLs.Gastric mucosal Bax protein expression was increased and Bcl-2 expression was decreased after C48/80 treatment,and ASLs ameliorated Bax and Bcl-2 expression.The extrusion process significantly augmented the effects of ASLs in a dosedependent manner.ASLEs at 200 mg/kg normalized mucus damage/secretion,C48/80-induced increases of mucosal myeloperoxidase activity(index of inflammation),xanthine oxidase,and malondialdehyde content(index of lipid peroxidation).The effects of ASLs on Bax and Bcl-2expression were also enhanced by extrusion.Furthermore,these effects of ASLEs at 200 mg/kg were similar to those of famotidine,a histamine H2-receptor antagonist commonly used to treat gastric ulcers.CONCLUSION:ASLEs prevented acute gastric mucosal lesion progression induced by C48/80,possibly by inducing mucus production,and reduced inflammation and oxidative stress in gastric mucosa through an anti-apoptotic mechanism.

Quantitative analysis of endangered Acanthopanax senticosus communities in Dongling Mountain of Beijing

Based on the survey of community plots,a quantitative analysis of endangered Acanthopanax senticosus communities in Dongling Mountain was performed with two way indicator species analysis(TWINSPAN),detrended correspondence analysis(DCA)and canonical correspondence analysis(CCA).The communities of A.senticosus were classified into 9 types by TWINSPAN and the results were validated by DCA.On the DCA graph,the first axis reflected the gradient of altitude and the second axis reflected the aspect and slope.Most of A.senticosus were distributed in the thick forests at a high altitude with little light.With the exception of being a dominant species of shrub layers in a few communities,A.senticosus has a relatively scarce distribution.In accordance with DCA,the results of CCA also show the trend that the distribution of A.senticosus communities varied along with the gradient change of environmental factors.Altitude and light are the main factors affecting A.senticosus growth.

Potential mechanism of Shugan Jieyu capsule in the treatment of hepatitis B based on network pharmacology

Objective:In this study,the effective components and related targets of Shugan Jieyu Capsule(SGJY)in the treatment of hepatitis B were determined to explore the mechanism of SGJY in the treatment of hepatitis B.Methods:In this study,the effective components and targets of SGJY,and the related targets of hepatitis B were searched,and obtained the targets of SGJY in the treatment of hepatitis B according to the principle of Venn diagram.To build a protein-protein interaction network,String database was used,Cytoscape(3.7.2)software was used for topology analysis,R(4.0.5)software was used for go analysis,KEGG pathway enrichment analysis,and study of putative signaling pathways to determine how they could work.Results:SZJY was used to predict a total of 11 Chinese herbal components and 85 associated targets for the treatment of hepatitis B.34 important targets were examined,including AKT1,EGFR,and 10 important pathways were examined,including proteoglycans in cancer and the PI3K/Akt signaling pathway.Conclusion:SGJY in the treatment of hepatitis B mainly inhibits the secretion of HBsAg and HBeAg by affecting PI3K-Akt and proteoglycans in cancer,and inhibits the progression of liver cancer.

刺五加扦插育苗技术

为探索刺五加(Acanthopanax senticosus)扦插育苗技术,对刺五加不同激素种类及其配比、不同基质对插穗生根的影响进行试验。结果表明,NAA浓度为50 mg/L、浸泡时间为1 h的生根率最高可达85.6%,平均生根数为11.2条,平均根长为4.7 cm。选用河沙+珍珠岩+腐殖质作为基质,可促进插穗生根。

Antidepressant effects of Radix et Caulis Acanthopanacis Santicosi extracts on rat models with depression in terms of immobile behavior

OBJECTIVE: To investigate the antidepressant effects of the Radix et Caulis Acanthopanacis Santicosi extracts in response to the tail suspension test(TST)and forced swim test (FST) in rats.METHODS:Male Wistar rats (9-week-old) wereorally administered the Radix et Caulis Acanthopanacis Santicosi extracts(30, 100 and 300 mg/kg) daily for 14 consecutive days. Serum corticosterone level was measured by using commercially available kit.The c-fos immunoreactivity was analyzed using immunohistochemistry methods.RESULTS:The effect of Radix et Caulis Acanthopanacis Santicosi extracts on the immobility time was significantly decreased at high-dose(300 mg/kg)treated group in the TST and FST(47.7%and69.6%,P<0.05), and decreased the hypothalamic-pituitary-adrenal(HPA) axis response to the stress,as in-dicated by an attenuated corticosterone response anddecreasedc-fosimmunoreactivity in the hippocampal and hypothalamic paraventricular nucleus region.CONCLUSION:Radix et Caulis Acanthopanacis Santicosi extracts have beneficial effects on depression behaviors and restore both altered c-fos expression and HPA activity.Thus, Radix et Caulis Acanthopanacis Santicosi extracts may be a novel agent for the treatmentof stress-related disorders.

刺五加嫩枝扦插插穗处理

为了提高刺五加(Acanthopanax senticosus)扦插生根率,就刺五加不同成熟度插穗、不同剪切方式、插穗上芽保留数量以及绿叶保留数量进行了试验。结果表明,当年生半木质化枝条的生根率为45.6%,生根所需时间比当年生木质化枝条生根时间提早9 d。插穗剪成15 cm一段,每段保证有2~3个芽,基部两面斜切成楔形,保留2片叶,每片叶片保留1/3~1/2,对刺五加插穗生根指标效果最佳,生根率最高可达85.4%。