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.

Eleutheroside B or E enhances learning and memory in experimentally aged rats

Eleutheroside B or E, the main component of Acanthopanax, can relieve fatigue, enhance memory, and improve human cognition. Numerous studies have confirmed that high doses of acetylcholine significantly attenuate clinical symptoms and delay the progression of Alzheimer＇s disease. The present study replicated a rat model of aging induced by injecting quinolinic acid into the hippocampal CA1 region. These rats were intraperitoneally injected with low, medium and high doses of eleutheroside B or E （50, 100, 200 mg/kg）, and rats injected with Huperzine A or PBS were used as controls. At 4 weeks after administration, behavioral tests showed that the escape latencies and errors in searching for the platform in a Morris water maze were dose-dependently reduced in rats treated with medium and high-dose eleutheroside B or E. Hematoxylin-eosin staining showed that the number of surviving hippocampal neurons was greater and pathological injury was milder in three eleutheroside B or E groups compared with model group. Hippocampal homogenates showed enhanced cholinesterase activity, and dose-dependent increases in acetylcholine content and decreases in choline content following eleutheroside B or E treatment, similar to those seen in the Huperzine A group. These findings indicate that eleutheroside B or E improves learning and memory in aged rats. These effects of eleutheroside B or E may be mediated by activation of cholinesterase or enhanced reuse of choline to accelerate the synthesis of acetylcholine in hippocampal neurons.

Neuroprotective effect of Eleutheroside B on 1-methyl-4-phenylpyridinium ion-induced apoptosis in PC12 cells

Apoptosis and viability of PC12 cells following 1-methyl-4-phenylpyridinium ion （MPP＋）-induced injury were monitored by flow cytometry, following Annexin V-propidium iodide double labeling, and 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay, respectively. The release of lactate dehydrogenase, superoxide dismutase activity and levels of malondialdehyde were determined by UV spectrophotometry. The changes in mitochondrial membrane potential and the intracellular concentration of calcium were determined by flow cytometry, and the activity of caspase-3 was monitored by western blot. According to cell viability and apoptosis studies, MPP＋-induced apoptosis in PC12 cells was inhibited in the presence of 10 tJg/mL of Eleutheroside B Our results indicate that the neuroprotective effect of Eleutheroside B, following MPP＋-induced apoptosis in PC12 cells, involves increasing the anti-oxidative stress capacity of cells, maintaining the high-energy state of mitochondrial membrane potential, reducing intracellular calcium concentration and inhibiting caspase-3 activity.

天然木脂素苷类成分eleutheroside E对膝骨性关节炎的治疗作用及其与MMPs相互作用方式的分析

本文旨在探讨天然木脂素苷类成分eleutheroside E(EE)对膝骨性关节炎模型的治疗作用并对EE的作用机制进行分析,明确EE与MMPs相互作用的位点及结合方式。实验采用ACLT方法建立兔膝骨关节炎模型,通过关节腔注射方式给予EE进行治疗干预,从动物整体水平对EE在实验性兔膝骨性关节炎的治疗作用进行综合评价。进一步运用分子对接技术和分子动力学方法,对EE与MMPs相互作用位点和结合方式进行分析。结果显示:EE能明显改善骨关节炎部位炎细胞浸润、纤维组织增生及软骨表层破坏等情况,并降低关节液中炎症介质(IL-1β和PGE 2)以及MMP-3和MMP-9的水平。分子对接和分子动力学实验发现,EE配体可结合于MMP-3和MMP-9催化位点的凹槽中,EE糖环上的多个羟基可与MMP-3和MMP-9受体中的多个氨基酸形成氢键,这些氢键对于配体的结合起到了重要作用。本研究为开发木脂素苷类母核结构新型MMPs抑制剂以及天然来源的骨性关节炎候选治疗药物奠定了一定基础。

<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.

Combination of Geniposide and Eleutheroside B Exerts Antidepressant-like Effect on Lipopolysaccharide-Induced Depression Mice Model

Objective To study the antidepressant-like effect and action mechanism of geniposide and eleutheroside B combination treatment on the lipopolysaccharide(LPS)-induced depression mice model.Methods Depression mice model was established by lipopolysaccharide(LPS)injection.Totally 48 mice were randomly divided into 6 groups(8 rats per group)according to a random number table,including normal,model,fluoxetine(20 mg/kg),geniposide(100 mg/kg)+eleutheroside B(100 mg/kg),geniposide+eleutheroside B+WAY 100635(0.03 mg/kg),geniposide+eleutheroside B+N-methyl-D-aspartic acid receptor(NMDA,75 mg/kg)groups,respectively.After continuous administration for 10 days,autonomic activity tests after 30 min of administration were performed on the 10th day.On the 11th day,except for the normal group,the mice in the other groups were intraperitoneally injected with LPS(1 mg/kg),and the behavioral tests were performed 4 h later.Enzyme linked immunosorbent assay was used to detect tumor necrosis factor alpha(TNF-α)and interleukin-1β(IL-1β)levels in mice serum.The mRNA expression of indoleamine 2,3-dioxygenase(IDO)and nuclear transcription factor(NF-κB)were detected by real-time quantitative polymerase chain reaction.Western-blot analysis was used to detect IDO and NF-κB protein expressions in hippocampus tissue.Results Compared with the normal group,a single administration of LPS increased the immobility time in the forced swimming test(FST)and tail suspension test(TST,P<0.01),without affecting autonomous activity.Compared with the model group,fluoxetine and geniposide+eleutheroside B administration significantly improved the immobility time of depressed mice in the FST and TST,decreased serum IL-1βcontent,inhibited the expression levels of NF-κB gene and protein in hippocampus tissues(P<0.05 or P<0.01).Compared with the model group,geniposide+eleutheroside B treatment significantly reduced serum TNF-αcontent and inhibited IDO mRNA and protein expressions in hippocampus(P<0.05 or P<0.01).In addition,NMDA partly prevented the inhibition of IDO mRNA expression by geniposide+eleutheroside B;NMDA and WAY-100635 also partly prevented the reduction of IL-1ßcontent induced by geniposide+eleutheroside B treatment(P<0.05 or P<0.01).Conclusions The combination of geniposide and eleutheroside B showed a certain antidepression-like effect.Its main mechanism of action may be contributed to inhibiting the activation of NF-κB,decreasing the proinflammatory cytokines such as TNF-α,IL-1β,and inhibiting in the neuroinflammatory reaction.Additionally,it also affects tryptophan metabolism,reduces the expression of a key enzyme of tryptophan metabolism,IDO.And this antidepressant-like effect may be mediated by 5-hydroxytryptamine and glutamate systems.

Study on External Application of Eleutherosides Ointment on Shenque(CV 8)for Resisting Sleep Deprivation

Objective： To investigate the efficacy of external application of Eleutherosides ointment on Shenque （CV 8） for treating sleep deprivation. Methods： Twenty four healthy young male volunteers at the age of 19-22 years old were randomly allocated to 4 groups （normal, model, treatment and control groups）. Sleep was deprived for 48 hours except the normal group. Self-made Eleutherosides compound preparation was applied externally on Shenque （CV 8）, to observe its influence on psychology and blood biochemical indices of cortisol and testosterone of the human body in sleep deprivation. Results： After sleep deprivation for 48 hours, an increment in cortisol concentration was lower in the Eleutherosides group （44.482±96.065 nmol/L） than in the control group （146.809 ±71.075 nmol/L）; an increment in the self-evaluated depression scale was also lower in the Eleutherosides group （2.833 ± 16.746） than in the control group （20.417 ±10.358）. There were significant differences （P〈0.05）. But, there was no significant difference between the treatment group and control group （P〉0.05） in the decrement of testosterone concentration and the increment of the anxiety scale. Conclusion： External application of Eleutherosides preparation on Shenque （CV 8） can regulate the stress reaction level and psychological endurance of the human body and produce an effect to resist sleep deprivation.

Identifi cation of targets and mechanisms for Eleutheroside E in the treatment of cancer

Eleutheroside E(EE)is a monomer compound isolated from Acanthopanax senticosus,which has functions of antifatigue,antioxidation,and immune regulation.However,the function of EE in inhibiting cancer has rarely been reported.In this study,we used bioinformatics to determine the role and mechanisms of EE in inhibiting breast cancer(BRCA),prostate cancer(PRAD),lung cancer(LUAD),and bladder cancer(BLCA).Finally,the MTT method was used to evaluate the inhibitory effect of EE on breast cancer MDA-MB-231 cells,and the target genes were determined by qRT-PCR.The results showed that the survival rate of MDA-MB-231 cells was reduced by 27.33%after 400μg/mL EE treatment.The results of qRT-PCR showed that EE down-regulated target genes in breast cancer cells,which was consistent with our predicted results.Moreover,our results indicated that EE may have impact on the transcription and translation of BLCA cells by targeting ADCY2,ADCY5 and JUN.In BRCA,we identified three targets for EE.In addition,EE affected DNA repair and the development of BRCA by affecting the expression of RAD51.In LUAD,we identified 9 targets for EE,8 of which were related to DNA repair.EE inhibited PRAD by targeting ADCY8.Our research was the first report on the targets and mechanisms of EE for suppressing cancer combined with clinical data.Our research also provides theoretical support for using EE to suppress cancer.