Preparation techniques, structural features, and bioactivities of Eucommia ulmoides polysaccharides: A review.

Eucommia ulmoides Oliv. (tu-chung), as a famous nature medical and edible plant, has the effect of tonifying liver and kidney, strengthening the function of the muscles and bones, and miscarriage prevention. Accumulating evidence has demonstrated that the polysaccharides from Eucommia ulmoides Oliv. (EUPs) are a kind of vital and representative biologically active macromolecules and have various health-promoting biological activities in vivo and in vitro, such as antioxidant activity, immunomodulatory activity, hypolipidemic and hypoglycemic activities, anti-inflammatory activities, anti-tumor activity, and among others. The review aims to comprehensively and systematically collate the recent research progress on extraction and purification methods, structural characteristics, biological activities, mechanism of action, structural modification, and toxicity of EUPs to support their therapeutic potential and health-care functions. New valuable insights for future research with EUPs were also proposed in the areas of structural characterization and pharmacological activities to promoting the development of therapeutic agents and functional foods.

Research Progress of Eucommia ulmoides Oliv and Predictive Analysis of Quality Markers Based on Network Pharmacology.

Du Zhong is a valuable Chinese medicinal herb unique to China. It is a national second- class precious protected tree, known as "plant gold", which has been used to treat various diseases since ancient times. The main active ingredients are lignans, phenylprophetons, flavonoids, iridoids and steroids and terpenoids, which have pharmacological effects such as lowering blood pressure, enhancing immunity, regulating bone metabolism, protecting nerve cells, protecting liver and gallbladder and regulating blood lipids. In this paper, a comprehensive review of Eucommia ulmoides Oliv. was summarized from the processing and its compositional changes, applications, chemical components, pharmacological effects, and pharmacokinetics, and the Q-marker of Eucommia ulmoides Oliv. is preliminarily predicted from the aspects of traditional efficacy, medicinal properties and measurability of chemical composition, and the pharmacodynamic substance basis and potential Q-marker of Eucommia ulmoides Oliv. are further analyzed through network pharmacology. It is speculated that quercetin, kaempferol, ß-sitosterol, chlorogenic acid and pinoresinol diglucoside components are selected as quality markers of Eucommia ulmoides Oliv., which provide a basis for the quality control evaluation and follow-up research and development of Eucommia ulmoides Oliv.

A review of "plant gold" Eucommia ulmoides Oliv.: A medicinal and food homologous plant with economic value and prospect.

Eucommia ulmoides Oliv. is an ancient and precious plant that has been used as medicine in China for more than 2000 years. Because its bark, leaves, seeds, and male flowers can be used in medicine, it plays an important role in medicine, food, chemical industry, and other fields, so it is also called "plant gold". 246 compounds have been isolated from E. ulmoides, which endow E. ulmoides with many unique pharmacological effects and make it wide to study in the fields of osteoporosis, hypertension, liver protection, and so on. Besides, E. ulmoides also has significant medicinal effects on anti-inflammatory, antioxidant, immunomodulation, and neuroprotection, and is often used in clinical compound medicines of traditional Chinese medicine. In addition to updating its ethnobotany, phytochemistry, pharmacology, and toxicology information, the economic botany of leaves, seeds, and male flowers was also introduced. It hopes hoping to fully understand this economically important Chinese medicine and provide a scientific basis for further development and utilization of E. ulmoides.

Dynamic changes of endophytic bacteria in the bark and leaves of medicinal plant Eucommia ulmoides in different seasons.

The bark and leaves of the Eucommia ulmoides Oliv. (E. ulmoides) have good medicinal value. Studies show endophytes play important roles in host medicinal plant secondary metabolite synthesis, with season being a key influencing factor. Therefore, we used 16 S rRNA to detect endophytic bacteria (EB) in E. ulmoides bark and leaves collected in winter, spring, summer, and autumn, and analyzed the contents of major active components respectively. The results showed that the species diversity and richness of EB of the E. ulmoides bark were higher than those of leaves in all seasons except fall. Among them, the higher species diversity and richness were found in the E. ulmoides bark in winter and spring. EB community structure differed significantly between medicinal tissues and seasons. Concurrently, the bark and leaves of E. ulmoides showed abundant characteristic EB across seasons. For active components, geniposidic acid showed a significant positive correlation with EB diversity and richness, while the opposite was true for aucubin. Additionally, some dominant EB exhibited close correlations with the accumulation of active components. Delftia, enriched in autumn, correlated significantly positively with aucubin. Notably, the impact of the same EB genera on active components differed across medicinal tissues. For example, Sphingomonas, enriched in summer, correlated significantly positively with pinoresinol diglucoside (PDG) in the bark, but with aucubin in the leaves. In summary, EB of E. ulmoides was demonstrated high seasonal dynamics and tissue specificity, with seasonal characteristic EB like Delftia and Sphingomonas correlating with the accumulation of active components in medicinal tissues.

Pharmacokinetics, anti-rheumatoid arthritis activity, and active ingredient contents of Eucommia ulmoides Oliv.

Eucommia ulmoides Oliv. is a deciduous tree which contains various chemical ingredients. The main objective was to document the active chemical ingredients of Eucommia ulmoides Oliv. and their metabolic profiles in vivo, with a view to providing an experimental and theoretical basis for clarifying the mechanisms underlying the pharmacological activity of Eucommia ulmoides Oliv. against rheumatoid arthritis. Eight main active constituents of Eucommia ulmoides Oliv. bark (pinoresinol glucopyranoside, aucubin, geniposidic acid, geniposide, genipin, chlorogenic acid, quercetin and betulinic acid) were quantified using high-performance liquid chromatography (HPLC). This paper additionally identified and characterized prototype metabolites via ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) combined with the Human Metabolome Database (HMDB) and literature comparisons. Ultra pressure liquid chromatography-mass spectrometry/ mass spectrometry (UPLC-MS/MS) was subsequently employed to quantify these components in blood over time and evaluate their pharmacokinetic characteristics. The anti-rheumatoid arthritis effects of genipin, pinoresinol glucopyranoside and their combinations were assessed using in vitro cellular assays. We identified and characterized a total of 53 ingredients from Eucommia ulmoides Oliv. bark and plasma samples, among which 20 were confirmed as prototype metabolites. Meanwhile, this paper derived and analyzed the metabolic cleavage pathway of 8 index ingredients. Six of these compounds displayed rapid entry into blood, with high plasma exposure and fast elimination rates. Data from the in vitro cellular assay showed that aucubin, pinoresinol glucopyranoside, genipin, and combinations of these compounds effectively inhibit MH7A cell proliferation, reduce NO release, and decrease inflammatory factor levels.

Feeding Laying Ducks Eucommia ulmoides oliv. Leaves Increases the n-3 Fatty Acids Content and Decreases the n-6: n-3 PUFA Ratio in Egg Yolk without Affecting Laying Performance or Egg Quality.

The objective was to determine the effects of supplementing duck diets with Eucommia ulmoides oliv. leaf powder (EUL). Laying ducks (n = 480) were randomly allocated into 4 experimental treatments and fed diets containing 0, 1, 2, or 4% EUL. Dietary inclusion of EUL had no effect (p > 0.05) on laying performance or egg quality, but linearly increased (p < 0.05) total plasma protein, globulin, and HDL-C concentrations with concurrent reductions (p < 0.05) in plasma concentrations of cholesterol and LDL-C. Eggs laid by ducks receiving EUL had yolks with linearly higher phenolic concentrations (p < 0.05) but lower cholesterol concentrations (p < 0.05). EUL supplementation in duck diets significantly reduced n-6: n-3 PUFA ratio by enriching n-3 fatty acids in yolks (p < 0.05) with no changes in n-6 PUFA (p >0.05).

Research progress on the protective effects of aucubin in neurological diseases.

CONTEXT: Aucubin (AU), an iridoid glycoside that is one of the active constituents of Eucommia ulmoides Oliv. (EUO) (Eucommiaceae), a traditional Chinese medicine, has been extensively studied in the management of neurological diseases (NDs). However, a comprehensive review of its effects and mechanisms in this regard is currently not available. OBJECTIVE: To compile the protective effects and mechanisms of AU in NDs and provide a basis for further research. METHODS: We used 'aucubin' as the 'All Fields' or 'MeSH' in PubMed, Web of Science and China National Knowledge Infrastructure without any limitation to search all relevant articles as comprehensively as possible; we selected the articles on AU treatment of NDs for summary. RESULTS: Studies reviewed herein reported that AU improved the symptoms or prognosis of Parkinson's disease, Alzheimer's disease, intracerebral haemorrhage, diabetic encephalopathy, epilepsy, anxiety and depression, and traumatic brain injury. The pharmacological mechanisms involved in repairing neuronal loss were postulated to include increasing γ-aminobutyric acid (GABA) content in the synapse, promoting differentiation of neural precursor cells into GABAergic neurons, providing antioxidant and anti-neuroinflammation activities, as well as enhancing autophagy and anti-apoptotic actions. DISCUSSION AND CONCLUSIONS: The protective effects of AU on some NDs have been confirmed. According to the pharmacological effects, AU is also highly likely to have protective effects on other NDs, which can be realized by further in vivo and in vitro basic research, and clinical trials. In the future, AU may be used for clinical prevention or treatment of patients with neurological diseases.

UPLC-MS analysis and network pharmacology-based investigation into the active ingredients and molecular mechanisms of anti-fatigue of male flowers with Eucommia ulmoides Oliv.

The male flowers of Eucommia ulmoides Oliv. (MFEU) was a natural product that could alleviate fatigue and accelerate fatigue alleviation. Nonetheless, the active ingredients and underlying pharmacological mechanisms remain unknown. This study aimed to decode the active ingredients and potential action mechanisms of MFEU in the therapy of anti-fatigue using an integrated UPLC-MS analysis, network pharmacology approach, and cell experiments. Characterizations of chemical constituents of MFEU extract were identified by UPLC-Q-TOF-MS. The corresponding drug targets were retrieved from the drug target database and used to construct the "composite-target-pathway" network. The Cytoscape was used to identify potential protein targets of these MFEU components, indicating that 24 anti-fatigue compounds in MFEU regulate 18 anti-fatigue-related targets in 10 signaling pathways. The 16 components of MFEU were verified at the cellular level. The results of cell experiments showed that MFEU extract (0.361 µg/ml), Caffeic acid, Deacetylasperulosidic acid, Naringenin, Acanthoside B, Geniposidic acid, Rutin, and Quercetin could promote testosterone secretion on Leydig cells at 50 µM. The MFEU extract and seven compounds in MFEU might play a role in anti-fatigue by participating in the regulation of testosterone secretion. Finally, the results of PCR analysis showed that MFEU promotes the secretion of testosterone, which is related to CYPIIa1 and 17ß-HSD, STAR in the signal pathway of testosterone synthesis. This study provides a basis for further exploring the anti-fatigue mechanism of MFEU, adopting the method of multi-compound and multi-target.

Assembly, Core Microbiota, and Function of the Rhizosphere Soil and Bark Microbiota in Eucommia ulmoides.

Medicinal plants are inhabited by diverse microbes in every compartment, and which play an essential role in host growth and development, nutrient absorption, synthesis of secondary metabolites, and resistance to biological and abiotic stress. However, the ecological processes that manage microbiota assembly and the phenotypic and metabolic characteristics of the core microbiota of Eucommia ulmoides remain poorly explored. Here, we systematically evaluated the effects of genotypes, compartment niches, and environmental conditions (climate, soil nutrition, and secondary metabolites) on the assembly of rhizosphere soil and bark associated bacterial communities. In addition, phenotypic and metabolic characteristics of E. ulmoides core microbiota, and their relationship with dominant taxa, rare taxa, and pharmacologically active compounds were deciphered. Results suggested that microbiota assembly along the two compartments were predominantly shaped by the environment (especially pH, relative humidity, and geniposide acid) and not by host genotype or compartment niche. There were 690 shared genera in the rhizosphere soil and bark, and the bark microbiota was mainly derived from rhizosphere soil. Core microbiota of E. ulmoides was a highly interactive "hub" microbes connecting dominant and rare taxa, and its phenotypic characteristics had a selective effect on compartment niches. Metabolic functions of the core microbiota included ammonia oxidation, nitrogen fixation, and polyhydroxybutyrate storage, which are closely related to plant growth or metabolism. Moreover, some core taxa were also significantly correlated with three active compounds. These findings provide an important scientific basis for sustainable agricultural management based on the precise regulation of the rhizosphere soil and bark microbiota of E. ulmoides.

Nocardiopsis eucommiae sp. nov., a novel endophytic actinomycete isolated from leaves of Eucommia ulmoides Oliv.

A novel Gram-stain-positive, endophytic actinobacterium, designated strain HDS5T, was isolated from leaves of Eucommia ulmoides Oliv. collected from Changde City, Hunan Province, PR China. Strain HDS5T produced yellowish oil green substrate mycelia on Gause's synthetic medium, which also carried yellowish oil green aerial hyphae, fragmenting into rod-shaped elements with smooth surfaces. Strain HDS5T grew at pH 5.0-11.0 (optimum, pH 7), at 20-40 °C (optimum, 28 °C) and in the presence of 0-8.0% NaCl (w/v; optimum, 0-1.0 %). Whole-cell hydrolysates contained meso-diaminopimelic acid as the diagnostic amino acid and no diagnostic sugars. The predominant fatty acids were iso-C16:0 and C18 : 1 ω9c. The menaquinones were MK-10(H2), MK-10(H4) and MK-10(H6). Strain HDS5T showed high 16S rRNA gene sequence similarity to Nocardiopsis prasina DSM 43845T (99.72 %), Nocardiopsis ganjiahuensis DSM 45031T (99.31 %), Nocardiopsis exhalans JCM 11759T (99.17 %), Nocardiopsis alba DSM 43377T (99.11 %), Nocardiopsis metallicus KBS6T (99.11 %), Nocardiopsis valliformis DSM 45023T (99.04 %), Nocardiopsis listeri NBRC 13360T (98.97 %), Nocardiopsis lucentensis DSM 44048T (98.83 %), Nocardiopsis terrae YIM 90022T (98.83 %) and <98.7 % similarities to other type strains. Phylogenetic analysis of 16S rRNA gene sequences and whole-genome sequences showed that strain HDS5T was closely related to N. prasina DSM 43845T. However, the average nucleotide identity based on blast and digital DNA-DNA hybridization values between them were determined to be 90.1 and 40.9 %, respectively, below the threshold of 95-96 and 70 % for the delineation of prokaryotic genomic species, suggesting that strain HDS5T represents a novel Nocardiopsis species. Furthermore, the morphological and physio-biochemical characteristics were sufficient to distinguish strain HDS5T from N. prasina DSM 43845T. Consequently, based on phenotypic and genotypic characteristics, strain HDS5T represents a new Nocardiopsis species, for which the name Nocardiopsis eucommiae sp. nov. is proposed. The type strain is HDS5T (=MCCC 1K06172T=JCM 34707T).

An alcohol extract prepared from the male flower of Eucommia ulmoides Oliv. promotes synoviocyte apoptosis and ameliorates bone destruction in rheumatoid arthritis.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease dominated by synovial hyperplasia and bone destruction. The male flower of Eucommia ulmoides Oliv. (EF) has been shown to exert effects on the inflammation caused by RA. However, how EF affects synoviocyte apoptosis and bone destruction on RA have not been investigated thoroughly. The effects of EF on apoptosis of human fibroblast-like synoviocytes-rheumatoid arthritis (HFLS-RA) cells, osteoclast differentiation of RAW264.7 cells, and bone destruction in a collagen-induced arthritis (CIA) model in rats were explored. METHODS: First, the main components of EF were identified by high-performance liquid chromatography. In vitro, we investigated the anti-proliferative and pro-apoptotic effects of EF on HFLS-RA cells by immunofluorescence assays, flow cytometry, real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and western blotting; we also investigated how EF influenced the differentiation of RAW264.7 cells into osteoclasts. In vivo, we used a rat model of CIA to investigate the effects of EF on anti-arthritis activity, toe swelling, Arthritis Score, serum levels of metabolic bone factors, and pathologic conditions. Micro-computed tomography was used to scan ankle joints. mRNA and protein expression of factors related to the nuclear factor-kappa B (NF-κB) pathway were determined by RT-qPCR and western blotting, respectively. RESULTS: EF inhibited synoviocyte proliferation and promoted apoptosis in a dose-dependent manner. EF inhibited osteoclast differentiation by inhibiting activation of the NF-κB pathway. EF reduced articular inflammation in CIA rats, inhibited the expression of pro-angiogenic factors, and delayed the destruction of articular cartilage and bone. Our data indicated that EF acted via a mechanism related to bone metabolism induced by the NF-κB pathway. CONCLUSIONS: EF exerts a potential therapeutic effect upon RA. Our research will help to elucidate the potential pharmacologic mechanisms associated with the beneficial effects of EF, and provide an experimental basis for EF application in clinical treatments.

Effects of total flavonoids from Eucommia ulmoides Oliv. leaves on polycystic ovary syndrome with insulin resistance model rats induced by letrozole combined with a high-fat diet.

ETHNOPHARMACOLOGICAL RELEVANCE: Eucommia ulmoides Oliv. leaves are the dry leaves of Eucommia ulmoides Oliv. Modern studies have shown that Eucommia ulmoides Oliv. leaves and its extracts have many pharmacological effects, such as regulating hypothalamus pituitary ovary (HPO) axis function, estrogen like effects, correcting insulin resistance (IR), regulating lipids, and reducing weight, which are consistent with the clinical manifestations in polycystic ovary syndrome (PCOS) patients. PCOS patients often have HPO axis disorder, low estrogen, high androgen, high IR complication rate, and obesity. Previous preclinical studies have shown that total flavonoids from Eucommia ulmoides Oliv. leaves (TFEL) can improve the imbalance in sex hormone secretion in perimenopausal animal models by regulating the function of the HPO axis. Thus, it is important to understand if flavonoids are the active parts of Eucommia ulmoides Oliv. leaves that interfere with polycystic ovary syndrome with insulin resistance (PCOS-IR), and determine the regulatory role they play in sex hormones and IR? AIM OF THE STUDY: Investigate the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway in the ovary and kisspeptin/insulin like growth factor/leptin receptor1/androgen receptor (Kiss1/IGF-1/LEPR/AR) in the HPO axis to determine the mechanism of TFEL intervention in a rat model of PCOS-IR model rats. MATERIALS AND METHODS: A rat model of PCOS-IR was established using a high-fat diet (49 d) combined with letrozole (1 mg/kg·d, for 28 d). Then, metformin (300 mg/kg·d) and TFEL (220 mg/kg·d, 110 mg/kg·d, and 55 mg/kg·d) were administered continuously for 21 days. At the end of the experiment, samples were taken and the related indexes were measured. RESULTS: TFEL reduced the body weight, Lee's index, ovarian index, ovarian area and ovarian volume, increased serum E2, SHBG levels and ISI, decreased serum levels of T, LEP, INS, and FBG (whole blood), and reduced the HOMA-IR in rats with PCOS-IR. TFEL downregulate Kiss1, IGF-1, and AR in the arcuate nucleus of hypothalamus, and upregulate Kiss1, downregulate IGF-1 and AR in the pituitary gland, and upregulate Kiss1, downregulate IGF-1, LEPR, and AR in the ovary of rats with PCOS-IR. TFEL could downregulate p-IRS-1Ser307, upregulate IRS-1, p-IRS-1Tyr895, PI3Kp85α, p-PI3Kp85α, AKT, p-AKT, and GLUT4 in the ovary, and ameliorated histopathological changes in the ovary and pancreas of rats with PCOS-IR. CONCLUSION: TFEL can inhibit ovarian hyperplasia, regulate disorders of glucose and lipid metabolism and improve the secretion of sex hormones, by regulating the expression of PI3K/AKT signaling pathway-related proteins in the ovary and Kiss1/IGF-1/LEPR/AR in the HPO axis.

Adsorption and desorption characteristics of polyphenols from Eucommia ulmoides Oliv. leaves with macroporous resin and its inhibitory effect on α-amylase and α-glucosidase.

BACKGROUND: Eucommia ulmoides Oliv. (EUO) was a traditional Chinese herb, its leaves were abundant in China, and polyphenol compounds were considered to be an important active ingredient in Eucommia ulmoides Oliv. leaves (EUOL). However, previous research mainly focused on compound identification and extraction process, there were few reported on the efficient enrichment process and biological activity evaluation of polyphenols in EUOL. METHODS: The adsorption and desorption characteristics of twelve different resins (HPD-100, HPD-300, HPD-600, D-3250, X-5, D-140, NKA-9, NKA-II, D-101, AB-8, S-8 and Polyamide) were investigated to develop an efficient method for the enrichment of polyphenol from EUOL, and the static adsorption, kinetics, isotherm and thermodynamics of the polyphenol from EUOL were analyzed. The eluted component was obtained through dynamic elution, and its main polyphenol compounds were detected by high-phase liquid chromatography (HPLC) and the inhibitory effects on the enzyme activity of α-amylase and α-glucosidase was also evaluated for different elution components. Meanwhile, the binding of main polyphenol compounds to enzyme was also evaluated. RESULTS: The selected resins (HPD-300, HPD-600, D-3250, X-5, D-140, NKA-9, D-101 and AB-8) showed adsorption patterns that fitted well to the pseudo second-order kinetics. The intra-particle diffusion model demonstrated that the diffusion of polyphenol compounds on these resins were divided into three processes. For HPD-300, HPD-600 and NKA-9, the Freundlich model better described the adsorption isotherm behavior than the Langmuir model, and the adsorption of polyphenol was a physical, exothermic, and spontaneous process. Subsequently, dynamic elution was performed yielding a higher polyphenol content in a 60% ethanol-water elution component, and it also exhibited a higher inhibitory effect on α-amylase and α-glucosidase activity. Furthermore, as the main polyphenol compounds, chlorogenic acid, rutin, quercetin and kaempferol were used to simulate the binding to the enzyme protein through molecular docking technology. The results showed that quercetin had a higher docking score for α-amylase, while rutin displayed superior binding to α-glucosidase. CONCLUSIONS: Therefore, polyphenols of EUOL could be enriched through macroporous resins and have the potential to be effective enzyme inhibitor.

Chemotype classification and biomarker screening of male Eucommia ulmoides Oliv. flower core collections using UPLC-QTOF/MS-based non-targeted metabolomics.

BACKGROUND: In the Chinese health care industry, male Eucommia ulmoides Oliv. flowers are newly approved as a raw material of functional food. Core collections have been constructed from conserved germplasm resources based on phenotypic traits and molecular markers. However, little is known about these collections' phytochemical properties. This study explored the chemical composition of male E. ulmoides flowers, in order to provide guidance in the quality control, sustainable cultivation, and directional breeding of this tree species. METHODS: We assessed the male flowers from 22 core collections using ultra-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) non-targeted metabolomics, and analyzed them using multivariate statistical methods including principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares discriminant analysis (OPLS-DA). RESULTS: We annotated a total of 451 and 325 metabolites in ESI+ and ESI- modes, respectively, by aligning the mass fragments of the secondary mass spectra with those in the database. Four chemotypes were well established using the ESI+ metabolomics data. Of the 29 screened biomarkers, 21, 6, 19, and 5 markers corresponded to chemotypes I, II, III, and IV, respectively. More than half of the markers belonged to flavonoid and amino acid derivative classes. CONCLUSION: Non-targeted metabolomics is a suitable approach to the chemotype classification and biomarker screening of male E. ulmoides flower core collections. We first evaluated the metabolite profiles and compositional variations of male E. ulmoides flowers in representative core collections before establishing possible chemotypes and significant biomarkers denoting the variations. We used genetic variations to infer the metabolite compositional variations of male E. ulmoides flower core collections instead of using the geographical origins of the germplasm resources. The newly proposed biomarkers sufficiently classified the chemotypes to be applied for germplasm resource evaluation.

Anti-neuroinflammatory effects of Eucommia ulmoides Oliv. In a Parkinson's mouse model through the regulation of p38/JNK-Fosl2 gene expression.

ETHNOPHARMACOLOGICAL RELEVANCE: Eucommia ulmoides Oliv., a Chinese medicinal herb called "Duzhong" from the bark of Eucommia ulmoides Oliv., has been shown to possess significant protective effects in Parkinson's disease (PD). However, the molecular mechanism remains unclear. AIM OF THE STUDY: In this study, we explored the anti-neuroinflammatory mechanisms of Duzhong on the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse model to elucidate the traditional medical theories with modern pharmacological methods and to provide a reference for further clarifying its mechanisms of action. MATERIALS AND METHODS: The representative components in Duzhong extract were identified by UPLC-Q-TOF/MS. Male C57BL/6J mice were intraperitoneally injected with MPTP to establish an in vivo PD model. The pole, rotarod, and grip strength tests were performed to evaluate the motor coordination ability of the PD mice. HPLC-ECD was used to detect the striatal levels of dopamine (DA), 3,4- dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). The expression of tyrosine hydroxylase (TH) was studied by immunohistochemistry (IHC) and Western blot assays. ELISA and Q-PCR were used examined the levels of proinflammatory cytokines in the serum and midbrain, respectively. Whole-transcriptome analysis of the midbrain was performed to explore the therapeutic effect of Duzhong on PD mice, and Q-PCR was then used to validate the differential gene expression changes in the PD mice treated with Duzhong. RESULTS: Ten compounds were identified from Duzhong extract. Duzhong significantly alleviated the behavioral impairments and dopaminergic neuron degeneration of PD mice, and inhibited the expression of proinflammatory cytokines. Whole-transcriptome analysis revealed nine oppositely regulated genes, and the Fosl2 gene was consistent with the trend of observed by RNA-seq. Furthermore, Duzhong downregulated mRNA expression of p38 and JNK, which are key upstream genes of Fosl2. CONCLUSIONS: Duzhong has promising therapeutic potential in PD mice, and its molecular mechanism is mediated by downregulating p38/JNK-Fosl2 gene expression to alleviate neuroinflammation.

Alcohol extract of eucommia ulmoides oliv. Promotes bone healing in rats with periapical periodontitis / 中国组织工程研究

BACKGROUND: Current research on Eucommia ulmoides Oliv. mainly focuses on its use in the treatment of osteoarthritis that Eucommia ulmoides Oliv. can enhance the healing ability of bone tissue. However, research on its bone repair ability in periapical periodontitis has not been reported. OBJECTIVE: To study the effect of alcohol extract of Eucommia ulmoides Oliv. on cathepsin K expression in periapical periodontitis rats. METHODS: Twenty-four Sprague-Dawley rats were divided into normal control group (n=4) and apical periodontitis group (n=20). In the periapical periodontitis group, a periapical periodontitis model was established after exposure of the dental pulp in the first molar of the right mandible. The normal control group did not deal with any treatment. After 4 weeks of feeding, four rats from each group were taken for micro-CT detection. Bone destruction was quantified to confirm whether the rat model of periapical periodontitis was successfully constructed. After 5 weeks of feeding, the remaining 16 rats with periapical periodontitis were equally randomized into alcohol extract group (given alcohol extract of Eucommia ulmoides Oliv. via intragastric administration, 5 mL/kg per day) and normal saline group (given the same dose of normal saline via intragastric administration every day). After 4 weeks of gavage, four mice from each group were selected to perform micro-CT examination. The ability of alcohol extract of Eucommia ulmoides Oliv. to repair periapical bone tissue was analyzed. First molars of the right mandible from the other four rats in each group were extracted to detect the expression of cathepsin K in the alveolar bone using immunohistochemical staining. RESULTS AND CONCLUSION: Micro-CT results showed that the rat model of periapical periodontitis was successfully constructed as there was a significant difference in the bone resorption volume between the normal control and apical periodontitis groups [(0.223±0.009) mm3 vs. (0.945±0.037) mm3, P=0.00]. After 4 weeks of gavage, the micro-CT results showed that the alcohol extract of Eucommia ulmoides Oliv. significantly reduced the bone resorption volume in the rat model of periapical periodontitis (P < 0.05). Immunohistochemistry results showed that the alcohol extract of Eucommia ulmoides Oliv. significantly inhibited the expression of cathepsin K, a marker of bone destruction, in the rat model of periapical periodontitis. Therefore, these findings indicate that the alcohol extract of Eucommia ulmoides Oliv. can inhibit the expression of cathepsin K and promote the healing of bone tissue in the rats with periapical periodontitis.

Screening of anti-complement active ingredients from Eucommia ulmoides Oliv. branches and their metabolism in vivo based on UHPLC-Q-TOF/MS/MS.

Eucommia ulmoides Oliv. (E. ulmoides) is a kind of plant with high medicinal value, there are known as the "gold plants". Some components and contents of barks and branches from E. ulmoides are similar, the barks are mainly used as medicine, but the branches have not been systematically studied and were discarded. In this paper, five fractions extracted from E. ulmoides branches were detected by the classical anti-complement activity assay in vitro. The n-butanol fraction of E. ulmoides branches showed excellent anti-complement activities with a CH50 value of 0.016 ±â€¯0.0014 mg·mL-1. A total of 76 compounds were identified from the n-butanol fraction, including 9 alkaloids, 18 organic acids, 22 lignans, 15 phenylethanoid glycosides and 12 other compounds. To further prove the anti-complement activity of potential active compounds, those compounds detectable in rat plasma after oral administration were tested by classical anti-complement activity assays. Genipin and pinoresinol 4-O-glucopyranoside had a certain complement inhibitory activity in the 17 potential anti-complements, their CH50 values were 0.050 ±â€¯0.0038 and 0.022 ±â€¯0.0018 mg·mL-1. UHPLC-Q-TOF/MS/MS was developed to profile and characterize the metabolites of genipin and pinoresinol 4-O-glucopyranoside in rat plasma. Twenty-one and seventeen metabolites were found, respectively. In summary, this study reported important clues for the further pharmacological and clinical studies of E. ulmoides branches. Meanwhile, it provided a practical strategy for rapid screening and identifying of in vivo anti-complement in traditional Chinese medicine.

Du-Zhong (Eucommia ulmoides Oliv.) Cortex Extract Alleviates Lead Acetate-Induced Bone Loss in Rats.

The purpose of this study was to evaluate the protective effect of Du-Zhong cortex extract (DZCE) on lead acetate-induced bone loss in rats. Forty female Sprague-Dawley rats were randomly divided into four groups: group I (control) was provided with distilled water. Group II (PbAc) received 500 ppm lead acetate in drinking water for 60 days. Group III (PbAc+DZCE) received 500 ppm lead acetate in drinking water, and given intragastric DZCE (100 mg/kg body weight) for 60 days. Group IV (DZCE) was given intragastric DZCE (100 mg/kg body weight) for 60 days. The bone mineral density, serum biochemical markers, bone histomorphology, and bone marrow adipocyte parameters were analyzed using dual-energy X-ray absorptiometry, biochemistry, histomorphometry, and histopathology, respectively. The results showed that the lumbar spine and femur bone mineral density was significantly decreased in PbAc group compared with the control (P < 0.05); however, this decrease was inhibited by the intake of Du-Zhong cortex extract (P < 0.05, vs. PbAc group; P > 0.05, vs. control and DZCE group). Serum calcium and serum phosphorus in the PbAc+DZCE group were greater than that in the PbAc group (P < 0.05). The PbAc group had higher ALP, osteocalcin, and RANKL than the control group (P < 0.01), and they were significantly lower in the PbAc+DZCE group compared with the PbAc group. There were no significant differences of ALP, osteocalcin, and RANKL among the PbAc+DZCE, control, and DZCE groups (P > 0.05). Serum OPG and OPG/RANKL ration were significantly higher in the PbAc+DZCE group than that in the PbAc group (P < 0.05). The bone histomorphometric analyses showed that bone volume and trabecular thickness in the femoral trabecular bone were significantly lower in the PbAc group than that in the control group, but those were restored in the PbAc+DZCE groups. The bone marrow adipocyte number, percent adipocyte volume per tissue volume (AV/TV), and mean adipocyte diameter were significantly increased in the PbAc group compared to the control (P < 0.01), and those were restored in the PbAc+DZCE group. The differences of those parameters between PbAc+DZCE, DZCE, and the control group were not significant. The results above indicate that the Du-Zhong cortex extract has protective effects on both stimulation of bone formation and suppression of bone resorption in lead-exposed rats, therefore, Du-Zhong cortex extract has the potential to prevent or treat osteoporosis resulting from lead expose.

Therapeutic effects and spectra-effect relationship of different extracts of Eucommia ulmoides on Parkinson’s disease mice / 中草药

Objective: To explore the therapeutic effects of different extracts of Eucommia ulmoides on Parkinson’s disease mice, as well as the relationship between ultra-high performance liquid chromatography (UPLC) fingerprint and treatment of Parkinson’s disease. Methods Through the mouse climbing test and the content of dopamine (DA) in the striatum of the brain, the therapeutic effect of different gradient ethanol extracts of E. ulmoides on Parkinson’s disease mice was observed. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to analyze the fingerprints of different extracts of E. ulmoides. Combined with the results of climbing rod test and dopamine content, partial least squares regression (PLSR) analysis was used to establish the pharmacodynamic relationship between E. ulmoides and Parkinson’s disease. Results The 50% and 75% ethanol extracts of E. ulmoides could significantly shorten the climbing time. The 75% ethanol extract of E. ulmoides significantly increased the striatum dopamine content in the brain. The results of PLSR analysis showed that ulmoside, liriodendrin, 5-hydroxymethylfurfural, caffeic acid in E. ulmoides were closely related to climbing rod and dopamine content of mice. Conclusion The ethanol extract of E. ulmoides has anti-Parkinson’s disease effect, and the effect is most significant with 75% alcohol extract. The compounds of ulmoside, liriodendrin, 5-hydroxymethylfurfural, caffeic acid may be the main active ingredients of E. ulmoides in the treatment of Parkinson’s disease.

Metabolite Profiles, Bioactivity, and HPLC Fingerprint of Different Varieties of Eucommia ulmoides Oliv.: Towards the Utilization of Medicinal and Commercial Chinese Endemic Tree.

Eucommia ulmoides Oliv. is widely regarded in China as a precious medicinal and commercial endemic tree. Due to cross-breeding or natural variation of E. ulmoides, the metabolite composition may vary significantly, making control of the medical quality difficult. In order to improve the rational development and utilization, the quality of seven varieties of E. ulmoides were evaluated based on metabolite profiles (total phenolic, total flavonoid, gutta-percha, aucubin, geniposidic acid, chlorogenic acid, geniposide, pinoresinol diglucoside, rutin, hyperoside, and astragalin), bioactivities (in vitro, in vivo antioxidant activities, and antibacterial activities) and HPLC fingerprint combined with chemometrics analysis. On this basis, the differences of medicinal parts (leaf and bark) were further carried out. For the traditional use of bark, Purple-leaf E. ulmoides was the most suitable. For the use of leaf, Qinzhong 1 and Purple-leaf E. ulmoides were appropriate. HPLC fingerprint analysis showed that significant differences in metabolite profiles exist among seven varieties of E. ulmoides. Combined with chemometrics analysis, seven varieties of E. ulmoides were divided into three groups from the use of leaf and bark. The analysis not only evaluated quality of seven varieties of E. ulmoides, but also could distinguish different varieties and different regions of origin. The results can provide theoretical basis for E. ulmoides resources utilization and cultivation of fine varieties.