To study the ameliorative effect of Eucommiae Cortex extract on spatial learning disabilities in APP/PS1 double-transgenic mice and explore its relationship with estrogen receptor β(ERβ)/c-Jun N-terminal kinase(JNK) signaling pathway, sixty 3-month-old male APP/PS1 mice were randomly divided into a model group, an anti-brain failure capsule group(0.585 g·kg~(-1)), a donepezil hydrochloride group(0.65 mg·kg~(-1)), and a Eucommiae Cortex extract group(1.3 g·kg~(-1)), and 15 C57BL/6 mice of the same genetic background were set as WT control group. The learning and memory ability of mice was assessed by the Morris water maze test(MWM), the passive avoidance test(PAT), and the novel object recognition test(NOR). The histomorphological and cellular ultrastructural features of the hippocampal region of the mice were observed by hematoxylin-eosin(HE) staining and transmission electron microscopy(TEM); the molecular docking validation of the key active ingredients and the key targets was performed by using AutoDock Vina software, and the immunohistochemical method(IHC) was used to detect the ERβ expression in the dentate gyrus(DG) area of mouse hippocampus. Western blot(WB) was utilized to detect the expression of ERβ, p-JNK, and JNK in mouse hippocampal area. Compared with those in the WT control group, the results of behavioral experiments showed that the latency of the mice in the model group was significantly increased, the number of platform traversals, and the target quadrant residence time were significantly decreased in the MWM. The evasion latency was significantly reduced, and the number of errors was significantly increased in the PAT. The index of recognition of novel objects was significantly reduced in the NOR. The results of HE staining indicated that the hippocampal area of mice in the model group showed a decrease in the number of neurons, disorganization of pyramidal cell arrangement, nucleus consolidation, and other changes. TEM results showed that some neuronal nuclei in the hippocampal area had a consolidated state, slightly thickened and aberrant nuclear membranes, and fewer intracytoplasmic nidus bodies; the IHC results showed that the expression of ERβ in the hippocampal DG area of the mice was reduced. The WB results showed that the ERβ expression in the hippocampal tissue was decreased, and the p-JNK/JNK level was elevated. Compared with the model group, the Eucommiae Cortex extract group showed a significant decrease in latency, and increase in number of platform traversals and target quadrant residence time in the MWM, a significant increase in evasion latency and decrease in number of errors in the PAT, and a significant increase in the index of recognition of novel objects in the NOR. In addition, there was an increase in the number of neurons in the hippocampal area of mice. The pyramidal cells tended to be arranged in an orderly manner; the nuclei of neurons in the hippocampal area were in a better state; the expression of ERβ in the hippocampal DG area of the mice was elevated; the expression of ERβ in the hippocampal tissue was elevated, and the level of p-JNK/JNK was reduced. The effects of donepezil hydrochloride group and anti-brain failure capsule on APP/PS1 mice in terms of behavioral, HE, and TEM indexes were similar to those of Eucommiae Cortex extract, and there was no significant difference between donepezil hydrochloride group and the model group in IHC and WB experiments, and the results of molecular docking indicated that the estrogen-like components in Eucommiae Cortex extract were tightly bound to ERβ. In conclusion, the binding of Eucommiae Cortex extract to estrogen receptors, regulation of ERβ expression, and activation of ERβ/JNK signaling pathway may be one of the key mechanisms by which it improves the learning and memory ability of APP/PS1 mice.
Animals ; Male ; Mice ; Mice, Transgenic ; Memory/drug effects* ; Mice, Inbred C57BL ; Estrogen Receptor beta/genetics* ; Eucommiaceae/chemistry* ; Alzheimer Disease/psychology* ; Amyloid beta-Protein Precursor/metabolism* ; Presenilin-1/metabolism* ; Humans ; MAP Kinase Signaling System/drug effects* ; Drugs, Chinese Herbal/administration & dosage* ; Hippocampus/metabolism* ; Maze Learning/drug effects* ; Learning/drug effects*

Trihelix transcription factors play important roles in plant light responses, growth and development, and stress responses. However, Trihelix has not yet been reported in Eucommia ulmoides. In this study, bioinformatics methods were used to comprehensively identify and analyze the expression patterns of the Trihelix gene family in E. ulmoides, aiming to provide a basis for further functional studies of EuGTs genes. A total of 9 Trihelix gene family members were identified in E. ulmoides, encoding proteins with 339 to 883 amino acids, with isoelectric points ranging from 5.13 to 9.39 and relative molecular weights between 36 992.06 and 97 871.61. Subcellular localization results showed that only EuGT-2 was localized in chloroplasts, while the others were located in the nucleus. The Trihelix gene family was categorized into six subfamilies: GT-1, GT-2, SH4, SIP1, GTγ, and GTδ. EuGTs were distributed among three subfamilies: SH4, GT-1, and GT-2, containing 1, 6, and 2 Trihelix proteins, respectively, with 2 to 17 exons. The promoters of EuGTs contained various cis-acting elements related to hormones, stress, photoperiod, and growth and development. Collinearity analysis revealed 5 collinear gene pairs between E. ulmoides and Arabidopsis thaliana, and 14 collinear gene pairs between E. ulmoides and Populus. Expression pattern analysis showed that EuGTs exhibited tissue-specific expression: EuGT-1, EuGT-2 had the highest expression levels in leaves, EuGT-4, EuGT-6, EuGT-9 had the highest transcriptional levels in marginal peel, and EuGT-5、EuGT-8 were predominantly expressed in the xylem. As leaves developed, EuGTs showed a trend of asynchronous changes. No significant differences in EuGTs expression were observed between male and female flowers, with high expression levels mainly during the induction stage of flowering. The qRT-PCR analysis indicated that most EuGTs genes were most highly expressed in the leaves of E. ulmoides, while EuGT-5 was highly expressed in the stems. Under 200 mmol·L~(-1) NaCl treatment, most EuGTs genes exhibited an initial increase followed by a decrease in expression, significantly responding to salt stress. This study provides important genetic resources for further exploration of EuGTs gene functions and germplasm innovation in E. ulmoides.
Plant Proteins/metabolism* ; Gene Expression Regulation, Plant ; Eucommiaceae/chemistry* ; Phylogeny ; Multigene Family/genetics* ; Gene Expression Profiling ; Transcription Factors/metabolism* ; Genome, Plant/genetics*

This study aims to develop the pre-column derivatization high performance liquid chromatography(HPLC) method for the determination of 16 kinds of amino acids in Eucommia ulmoides leaves, and compare the content of amino acids in the leaves harvested at different time and under leaf-oriented cultivation mode(LCM) and arbor forest mode(AFM). The HPLC conditions are as below: phenyl isothiocyanate(PITC) as pre-column derivatization agent, Agilent ZORBAX C_(18 )column(4.6 mm×250 mm, 5 μm), mobile phase A of acetonitrile-water(80∶20), mobile phase B of 0.1 mol·L~(-1) sodium acetate solution-acetonitrile(94∶6), gradient elution, flow rate of 1.0 mL·min~(-1), injection volume of 5 μL, column temperature of 40 ℃, and detection wavelength of 254 nm. The HPLC profile indicated well separation of 16 kinds of amino acids and the amino acid content in E. ulmoides leaves was up to 16.26%. In addition, the amino acid content in leaves of E. ulmoides under LCM was higher than under AFM. The amino acid content varied with the harvesting time. Through orthogonal partial least squares discriminant analysis, the amino acids of E. ulmoides under LCM and AFM were compared, which can distinguish the leaves under LCM from those under AFM. Principal component analysis was applied to comprehensively score the amino acids of E. ulmoides leaves. The results showed that the score of leaves under LCM was higher than that under AFM. Nutritional evaluation results indicated that the proteins in E. ulmoides leaves belonged to high-quality vegetable proteins. The established method for the determination of amino acid content is reliable. With the amino acid content as index, the leaf quality of E. ulmoides under LCM is better than that under AFM. This study lays a theoretical basis for the promotion of LCM for E. ulmoides and the development of medicinal and edible products from E. ulmoides leaves.
Amino Acids/metabolism* ; Eucommiaceae/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Plant Leaves/chemistry*

Lignans derived from Eucommia ulmoides Oliver (Eucommia lignans) inhibit the progression of inflammatory diseases, while their effect on the progression of diabetic nephropathy (DN) remained unclear. This work was designed to assess the function of Eucommia lignans in DN. The major constituents of Eucommia lignans were analyzed by UPLC-Q-TOF-MS/MS. The binding between Eucommia lignans and aldose reductase (AR) was predicted by molecular docking. Eucommia lignans (200, 100, and 50 mg·kg^-1) were used in model animals to evaluate their renal function changes. Rat glomerular mesangial cells (HBZY-1) were transfected with sh-AR, sh-AMPK, and oe-AR in the presence of high glucose (HG) or HG combined with Eucommia lignans to evaluate whether Eucommia lignans affected HG-induced cell injury and mitochondrial dysfunction through the AR/Nrf2/HO-1/AMPK axis. Eucommia lignans significantly attenuated the progression of DN in vivo. Eucommia lignans notably reversed HG-induced upregulation of inflammatory cytokines and mitochondrial injury, while downregulating the levels of Cyto c, caspase 9, AR, and NOX4 in HBZY-1 cells. In contrast, HG-induced downregulation of Nrf2, HO-1 and p-AMPKα levels were abolished by Eucommia lignans. Meanwhile, knockdown of AR exerted similar therapeutic effect of Eucommia lignans on DN progression, and AR overexpression reversed the effect of Eucommia lignans. Eucommia lignans alleviated renal injury through the AR/Nrf2/HO-1/AMPK axis. Thus, these findings might provide evidence for the use of Eucommia lignans in treating DN.
Animals ; Rats ; AMP-Activated Protein Kinases/genetics* ; Diabetes Mellitus ; Diabetic Nephropathies/prevention & control* ; Eucommiaceae/metabolism* ; Lignans/therapeutic use* ; Molecular Docking Simulation ; NF-E2-Related Factor 2/metabolism* ; Tandem Mass Spectrometry

Elucidate the mechanism of Eucommiae Cortex in treatment of hypertension, to provide the basis for further research and development of Eucommiae Cortex. Our study using the entity grammar systems inference rules to analyse the interactions of chemical constituents of Eucommiae Cortex and disease target proteins at the molecular level, and got a biological network of Eucommiae Cortex anti-hypertension which inciude 602 nodes and 2 354 edges. We got 3 treatment of hypertension pathways of Eucommiae Cortex by analyzing biological network, that is, by inhibition of vascular remodeling to improve the deterioration of hypertension, reduce activity of polymorphism genetic genes related to essential hypertension, inhibition of carbonic anhydrase 1 to maintain the osmotic pressure, Eucommiae Cortex play the role of anti-hypertension.
Animals ; Antihypertensive Agents ; administration & dosage ; chemistry ; Databases, Factual ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Essential Hypertension ; Eucommiaceae ; chemistry ; Gene Regulatory Networks ; drug effects ; Humans ; Hypertension ; drug therapy ; genetics ; metabolism ; Signal Transduction ; drug effects

OBJECTIVETo introduce the hairy root of Eucommia ulmoides, and screen put the excellent roots.

METHODHairy roots were obtained by infecting the different explants of E. ulmoides. Aucubin content was detected by HPLC.

RESULTThe best transformation condition was embryonic axis infected by twenty minutes bacillus water dip-dye and three-days co-culture. Liquid culture was better than solid culture, and advantageous to accumulating of biomass and medical contents of hairy root. The E1 was the excellent roots.

CONCLUSIONHairy root of E. ulmoides will be useful for the production of active components in aucubin.

Culture Media ; metabolism ; Eucommiaceae ; chemistry ; growth & development ; metabolism ; Glucosides ; analysis ; metabolism ; Iridoid Glucosides ; Iridoids ; analysis ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; Tissue Culture Techniques

OBJECTIVETo analyze and compare the cell growth and accumulation of flavonoids and chlorogenic acid in the callus and suspension cell of Eucommia ulmoides.

METHODThe callus induced from the leaf of E. ulmoides seedlings were suspended in liquid medium. The time courses of cell growth and yields of flavonoids and chlorogenic acid were studied.

RESULTThe highest contents of flavonoids and chlorogenic acid in the callus were 13.46, 1.712 mg x g(-1), respectively, while the contents of these two secondary metabolites were 16.63, 3.93 mg x g(-1) in suspension cell culture correspondingly.

CONCLUSIONComparing with callus, the suspension cell showed a short growth period and high growth rate with a remarkable high content of flavonoids and chlorogenic acid.

Cell Culture Techniques ; Cells, Cultured ; Chlorogenic Acid ; analysis ; metabolism ; Eucommiaceae ; chemistry ; metabolism ; Flavonoids ; analysis ; metabolism

OBJECTIVETo explore the pharmacodynamic and pathological mechanism of eucommia ulmoides oliv in improving erectile function.

METHODSThirty male diabetic rats were randomly divided into three groups: group A (n = 10, escipient group), group B (n = 10, sildenafil group), group C (n = 10, eucommia ulmoides oliv group) and group D (n = 10, the normal control group). After gavage for four weeks, the catching behaviors of all rats were observed, and ultrastructure of myelinated nerve fibers in penile tissue was examined by transmission electron microscope. The expression of neuronal nitric oxide synthase (nNOS) in penile tissues was examined by two steps immunohistochemistry method.

RESULTSCompared with group A, catching frequency of the rats in group C was notably increased (P < 0.05) and the expression of nNOS in penile tissue was significantly (P < 0.001). The examination by transmission electron microscope showed that in the rats' penile tissue of group A, myelinated nerve fibers were irregularly arranged and partially degenerated, and myelin sheaths lamella were splited and exhibited vacuoles or network forms. In group C, there were regular arrangements of myelinated nerve fibers, in which the formation of lamella was clear.

CONCLUSIONBy remitting the impairement of myelinated nerve fibers and enhancing the expression of nNOS in penile tissue, eucommia ulmoides oliv can improve erectile function of diabetic rats.

Animals ; Behavior, Animal ; drug effects ; Diabetes Mellitus, Experimental ; pathology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Eucommiaceae ; Female ; Immunohistochemistry ; Male ; Microscopy, Electron, Transmission ; Nerve Fibers, Myelinated ; ultrastructure ; Nitric Oxide Synthase ; metabolism ; Penis ; drug effects ; enzymology ; innervation ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of aqueous abstract from eucommia ulmoides oliv on the activity of superoxide dismutase (SOD) and alpha-actin expression in the penile tissues of rats with diabetes mellitus (DM) in vitro.

METHODSA diabetes model was established by administration of alloxun twice to Sprague Dawley (SD) rats. Ten diabetic and 10 normal rats were randomly selected and the penile strips of each rat were divided into four equal shares and cultured in two groups, a eucommia ulmoides oliv coculture group (Group A, further dicided into 1 microg/ml, 10 microg/ml and 100 microg/ml subgroups) and a control group (Group B). Seven days later, the activity of SOD in the culture medium was detected by spectrophotometry, and the levels of micro-actin expression in the penile tissues were examined with the immunohistochemical method.

RESULTSCompared with Group B, the activity of SOD in the culture medium in athe 10 and 100 microg/ml subgroups was notably elevated (P < 0.01), and the numbers of immunoreactive positive cells of alpha-actin in the penile tissues remarkably increased (P < 0.01).

CONCLUSIONThe activity of SOD and alpha-actin expression in the penile tissues of diabetic rats in vitro can be increased by eucommia ulmioides oliv.

Actins ; biosynthesis ; Alloxan ; Animals ; Diabetes Mellitus, Experimental ; metabolism ; Dose-Response Relationship, Drug ; Eucommiaceae ; Male ; Penis ; drug effects ; metabolism ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; drug effects ; metabolism

This study was designed to test the effect of Korean traditional tea materials on oxygen-free radical metabolism in lead (Pb) -administered rats. Male rats were divided into normal, Pb-control (Pb-Con) and Pb-water extract of green tea (Camellia sinensis; GT) , persimmon leaf (Diospyros kaki; PL) , safflower seed (Carhamus tinctorius: SS) , Du-Zhong (Eucommia ulmoides; EU) groups, respectively. Pb intoxication was induced by administration of lead acetate (25 mg/kg. B.W., oral) weekly. The extract was administered based on 1.26 g of raw material/kg B.W./day for 4 weeks. When the GT, PL, SS and EU were supplemented to the Pb-administered rats, hepatic lipid peroxide levels were significantly lower compared to the Pb-Con group. Hepatic cytochrom P-450 content and aminopyrine N-demethylase activity was lower in the Pb-Con group than in the normal group, whereas xanthine oxidase activity was significantly elevated in Pb-administered rats. The water extract of GT, PL, SS and EU supplementation attenuated changes in enzyme activities generating reactive oxygen species in the liver. Hepatic superoxide dismutase, catalase and glucose 6-phosphate dehydrogenase activities were significantly higher in the Pb-Con group than in the normal group, while monoamine oxidase activity also tended to increase in the Pb-administered rats. However, glutathione peroxidase and glutathione S-transferase activities, and glutathione content significantly decreased through Pb intoxication. The supplementation of GT, PL, SS and EU induced alleviation changes of hepatic antioxidant enzyme activity.
Aminopyrine N-Demethylase ; Animals ; Carthamus tinctorius ; Catalase ; Diospyros ; Eucommiaceae ; Glucose ; Glutathione ; Glutathione Peroxidase ; Glutathione Transferase ; Humans ; Liver ; Male ; Metabolism ; Monoamine Oxidase ; Oxidoreductases ; Oxygen* ; Rats* ; Reactive Oxygen Species ; Superoxide Dismutase ; Tea* ; Water* ; Xanthine Oxidase