Eucommia Ulmoides Oliv Research Articles

Kaempferol promotes osteogenic differentiation in bone marrow mesenchymal stem cells by inhibiting CAV-1.

Our study focused on the effects and molecular mechanisms of kaempferol, a major active component of Eucommia ulmoides Oliver (EUO), on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Target molecules for EUO, osteoarthritis, and osteogenic differentiation were identified through network pharmacology analysis. BMSCs were isolated and treated with various concentrations of kaempferol. Optimal concentration was determined through MTT assays. Osteogenic differentiation was assessed using alkaline phosphatase (ALP) and Alizarin Red S staining, while osteogenic markers (Collagen I, RUNX2, and OPN) and CAV-1 expression were analyzed using RT-qPCR and Western blot. The effects of combined treatment with kaempferol and an overexpression vector for CAV-1 (oe-CAV-1) on osteogenic differentiation were also observed. Network pharmacology analysis identified kaempferol as the primary active component influencing CAV-1 targeted in subsequent experiments. It was found that 10 µM kaempferol was optimal for treating BMSCs. Post-treatment, significant increases in ALP activity and calcium deposition were observed, along with elevated expression of osteogenic markers, and decreased CAV-1. Overexpression of CAV-1 significantly reversed the promotive effects of kaempferol on BMSC osteogenic differentiation, effectively inhibiting the process. Collectively, kaempferol promotes osteogenic differentiation in BMSCs by inhibiting CAV-1 expression.

Eucommia ulmoides Oliv. leaf extract: Effects on growth, antioxidant capacity, and lipid metabolism in Red swamp crayfish (Procambarus clarkii)

The addition of Eucommia ulmoides Oliv. leaf extract (ELE) enhances growth, immunity and antioxidant capacity in crustaceans, which has not been studied on the red swamp crayfish (Procambarus clarkii). Hence, this study aimed to investigate the effect of ELE on the growth, antioxidant capacity, digestive enzymes activities, and lipid metabolism of red swamp crayfish. A total of 240 individuals of red swamp crayfish were randomly assigned to four groups. Each group was fed with feed a diet supplemented with varying doses of ELE: 0 g/kg (CK), 0.5 g/kg (T2), 1.0 g/kg (T3), and 1.5 g/kg (T4). After 30 days, the results indicated that T2 group exhibited a significantly higher final average weight, weight gain rate, and specific growth rate compared to the CK (P<0.05). Significantly lower feed conversion ratios than the CK (P<0.05) were observed in the T2 and T3 groups. For antioxidant indicators, hepatopancreas superoxide dismutase (SOD) activity was observed to significantly increased in the T3 and T4 groups, while levels of malondialdehyde (MDA) in which were significantly reduced in hepatopancreas and hemolymph (P<0.05). Regarding biochemical indicators and intestinal function, significant enhancement of lipase (LPS) activity was observed in the T2 group, which decreased in the T4, compared to the CK (P<0.05). Moreover, significant decrease of the glucose (GLU), total triglyceride (TG), and total cholesterol (TC) content of the hemolymph (P<0.05) were observed in the T4 group. Transcriptome analysis revealed that differentially expressed genes in the T4 group were primarily enriched in peroxisome proliferator-activated receptor (PPAR), and lipid metabolism pathways. Furthermore, it seems that ELE supplements significantly upregulated the expression of apoa-II, acaox1, acox3, me1, and plin1 in the PPAR signaling pathway to regulate lipid metabolism. In conclusion, 0.5 g/kg and 1.5 g/kg ELL addition amount can enhance growth performance, antioxidant capacity, and lipid metabolism of red swamp crayfish.

Significant performance enhancement of Zn-ion hybrid supercapacitors based on microwave-assisted pyrolyzed active carbon via synergistic effect of NaHCO3 activation and CNT networks

Zinc-ion hybrid supercapacitors (ZIHSCs) represents a promising technological approach for large-scale energy storage with the combined advantages of supercapacitors and zinc-ion batteries. Unfortunately, it is still challengeable to quickly fabricate low-cost, high-performance carbonaceous cathode materials at relatively low temperature. To address such issues, herein, taking waste Eucommia ulmoides Oliver (EUO) wood as an example, we present a novel microwave-assisted carbonization (MWC) approach at relatively low temperature to quickly prepare active carbon, and we present a synergistic strategy to significantly enhance the electrochemical performance by introducing sodium bicarbonate activation (SA) and constructing conductive carbon nanotubes (CNT) networks. The MWC-SA@CNT hybrid exhibits outstanding specific capacitance of 344.2 F/g at 0.2 A/g within three-electrode system, much better than conventional high-temperature pyrolyzed AC, MWC carbon, and MWC-SA carbon. The superior performance of MWC-SA@CNT can be attributed to the synergistic effect of its large specific surface area of 1102.7 m2/g, high mesoporous percentage of 53.5%, and rich –OH and CO groups due to microwave-assisted carbonization and sodium bicarbonate activation, and rich electron transport paths due to the presence of CNT networks. Furthermore, ZIHSCs assembled by MWC-SA@CNT cathode could delivers impressive performance with excellent capacity (194.37 mA h/g at current density of 1 A/g), energy density (142.30 Wh/kg), and durability (capacitance retention rate of 97.65% after 5000 cycles). This work offers a rapid and low-temperature method for preparing wood-based active carbon with rich nanopores and strong conductivity to improve performance of Zinc ion storage.

Polysaccharides from Eucommia ulmoides Oliv. leaves alleviates alcohol-induced mouse brain injury and BV-2 microglial dysfunction

Acute alcohol intoxication is a harmful clinical condition characterized by behavioral and neurological symptoms, for which few effective therapies are available at present. Dysfunction of microglial BV-2 cells has been reported to be associated with acute alcohol-induced brain injuries. In the present study, the protective effects of Eucommia ulmoides Oliv. leaves polysaccharides (EULP) on acute alcoholic brain injury and microglial dysfunction were investigated. 14-day pretreatment of EULP significantly attenuated neurobehavioral deficit and neurotransmitter damage in the brain tissue of mice caused by acute alcohol exposure. Additionally, EULP regulated the metabolic disorder of brain tissue. Consistently, it was shown that EULP pretreatment significantly improved alcohol-induced phagocytosis decrease, oxidative stress and inflammation in BV-2 cells. Therefore, EULP may be proposed and employed as a potential therapeutic agent for alcohol-induced brain damage.

Cloning and functional characterization of the peptide deformylase encoding gene EuPDF1B from Eucommia ulmoides Oliv

Peptide deformylase can catalyse the removal of formyl groups from the N-terminal formyl methionine of the primary polypeptide chain. The peptide deformylase genes of a few herbaceous plants have been studied to some extent, but the peptide deformylase genes of woody plants have not been studied. In this study, we isolated EuPDF1B from Eucommia ulmoides Oliv. The full-length sequence of EuPDF1B is 1176 bp long with a poly-A tail and contains an open reading frame of 831 bp that encodes a protein of 276 amino acids. EuPDF1B was localized to the chloroplast. qRT‒PCR analysis revealed that this gene was expressed in almost all tissues tested but mainly in mature leaves. Moreover, the expression of EuPDF1B was enhanced by ABA, MeJA and GA and inhibited by shading treatment. The expression pattern of EuPDF1B was further confirmed in EuPDF1Bp: GUS transgenic tobacco plants. Among all the transgenic tobacco plants, EuPDF1Bp-3 showed the highest GUS histochemical staining and activity in different tissues. This difference may be related to the presence of enhancer elements in the region from − 891 bp to − 236 bp of the EuPDF1B promoter. In addition, the expression of the chloroplast gene psbA and the net photosynthetic rate, fresh weight and height of tobacco plants overexpressing EuPDF1B were greater than those of the wild-type tobacco plants, suggesting that EuPDF1B may promote the growth of transgenic tobacco plants. This is the first time that PDF and its promoter have been cloned from woody plants, laying a foundation for further analysis of the function of PDF and the regulation of its expression.

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, β-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.

Polysaccharides from Eucommia ulmoides Oliv. Leaves Alleviate Acute Alcoholic Liver Injury by Modulating the Microbiota-Gut-Liver Axis in Mice.

The interplay among gut microbiota, intestines, and liver is crucial in preventing acute alcoholic liver injury. In this study, the hepatoprotective potential of polysaccharides from Eucommia ulmoides Oliv. leaves (EULP) on acute alcoholic liver injury in Kunming male mice was investigated. The structural features suggested that the EULP appeared as a heterogeneous mixture of polysaccharides with a molecular weight of 186132 Da. A 14-day pretreatment of EULP ameliorated acute alcoholic-induced hepatic inflam mation (TNF-α, IL-6, and IL-10), oxidative stress (GSH, SOD, and T-AOC), and liver damage (ALT and AST) via enhancing intestinal barrier (Occludin, Claudin 1, and ZO-1) and modulating microbiome, which subsequently inhibiting endotoxemia and balancing the homeostasis of the gut-liver axis. EULP restored the composition of intestinal flora with an increase in the relative abundance of Lactobacillaceae and a decrease in Lachnospiraceae and Verrucomicrobiaceae. Notably, prolonged EULP pretreatment (14 days) but no single gavage of EULP achieved excellent hepatoprotection. These findings endorsed the potential of EULP as a functional food for mitigating acute alcoholic-induce d liver damage, attributed to its anti-inflammatory, antioxidant, and prebiotic properties facilitated by the microbiota-gut-liver axis.

Assessing the quality ecology of endemic tree species in China based on machine learning models and UPLC methods: The example of Eucommiaulmoides Oliv.

Climate change significantly affects the suitability distributions of natural populations, especially relict species such as Eucommia ulmoides Oliv. (E. ulmoides). However, the driving eco-factors of the distribution of E. ulmoides and the relationship between geo-distribution patterns and quality are still unclear. Here, a new approach was established by combining machine learning models (MaxEnt, Random Forest, and Biomod2 model), ultra-performance liquid chromatography (UPLC) and spatial analysis to address this question. The results reveal that temperature is the most critical variable influencing the distribution of E. ulmoides including Bio1, Bio10, and Bio11, followed by precipitation. Multi-model predictions of suitable habitats for E. ulmoides cover an area ranging from 113.33 × 104 km2 to 285.98 × 104 km2, mainly distributed in the mountainous areas of southwestern China, including the eastern regions of the Dalou, Wuling, and Qinling-Daba Mountains. Under different future scenarios, suitable habitat shifted northward by 184.90–424.88 km, and the contraction area is greater than the expansion area, with the difference ranging from 49.28 × 104 km2 to 98.39 × 104 km2. Eight common components (e.g., chlorogenic acid and pinoresinol doglucoside) and two endemic components (caffeic acid and geniposidic acid) were found in the leaves of the northern and southern medicinal districts (SDM and NMD). The contents of eight key secondary metabolites of E. ulmoides leaves were better distributed in SDM than in NDM. Among these, Xinyang in Henan, Lu'an and Fuyang in Anhui, and the southeastern region of Zhejiang are suitable growing areas for the high-quality cultivation of E. ulmoides, which deserve special attention. The seasonal precipitation (Bio15) was negatively correlated with chlorogenic acid and pinoresinol diglucoside while longitude significantly positively correlated with pinoresinol diglucoside. Overall, this study provides a novel assessment pattern for quality assessment and effective protection of E. ulmoides.

Quality and staling characteristics of wheat bread improved by adding Eucommia ulmoides oliver leaf micropowder

The wheat flour partially replaced by Eucommia ulmoides Oliver leaf micropowder (EUOLMP) was used to make bread, and the textural properties, moisture distribution, staling characteristics and internal structure of the resulting bread were analyzed to explore the effect of EUOLMP on the quality of wheat bread. The substitution level of EUOLMP had significant effects on the quality and staling characteristics of fresh bread. When 0.5%–1% of wheat flour was replaced by EUOLMP, the fresh bread gained higher specific volume, softer texture, improved porosity, and increased proportion of tightly bound water. The Differential Scanning Calorimetry (DSC) and X-ray diffraction (XRD) analysis demonstrated that the retrogradation enthalpy and relative crystallinity of bread decreased with the EUOLMP addition, which indicated that EUOLMP inhibited starch coagulation and recrystallization in bread and delayed the staling of bread. The EUOLMP substitution for wheat flour (at a 1% substitution level) could also obtain favorable sensory evaluation.

Chlorogenic acid ameliorates heart failure by attenuating cardiomyocyte ferroptosis

ObjectiveTo elucidate the effects of chlorogenic acid (CGA), a bioactive polyphenol compound prevalent in traditional Chinese medicine and various foods, including Lonicera japonica Thunb. (Jin Yin Hua), Eucommia ulmoides Oliv. (Du Zhong Ye), tea, and coffee, on cardiomyocyte ferroptosis and heart failure. MethodsWe assessed the effect of CGA on cardiac function using a mouse model of heart failure induced by transverse aortic constriction (TAC). These indicators included the left ventricular ejection fraction (LVEF), fractional shortening (LVFS), end-systolic volume (LVESV), end-diastolic volume (LVEDV), end-systolic diameter (LVESD), and end-diastolic diameter (LVEDD). An isoprenaline hydrochloride (ISO)-induced H9c2 cardiomyocyte cell model was also established, and the cells were treated with various concentrations of CGA. To assess the effect of CGA on ferroptosis in cardiomyocytes, we measured cell viability and evaluated the levels of intracellular reactive oxygen species (ROS), ferrous ions (Fe2+), and lipid peroxidation using fluorescent staining. To clarify the ferroptosis signaling pathway regulated by CGA, western blotting was used to examine the expression of ferroptosis biomarkers, specifically solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4), in H9c2 cardiomyocytes and mouse myocardial tissues. ResultsCGA significantly enhanced cardiac performance indices such as LVEF, LVFS, LVESV, LVEDV, LVESD, and LVEDD. H9c2 cardiomyocytes exposed to ISO showed decreased cell viability and increased ROS levels, Fe2+ content, and lipid peroxidation levels. However, CGA treatment significantly ameliorated these changes. Additionally, in both H9c2 cardiomyocytes and myocardial tissue obtained from mice with TAC, CGA increased the expression of ferroptosis-related proteins, including SLC7A11 and GPX4. ConclusionCGA has the potential to enhance cardiac function and diminish lipid peroxidation and ROS levels in cardiomyocytes via the SLC7A11/GPX4 signaling pathway. This process alleviates ferroptosis in cardiomyocytes. These results provide new insights into the clinical use of CGA and the management of heart failure.

Preparation of Eucommia ulmoides Oliver wood derived activated carbons by combined microwave hydrothermal pretreatment and microwave pyrolysis as electrode materials for super capacitive performance

In this work, the activated carbon was prepared from the Eucommia ulmoides Oliver (EUO) wood by a two-step method. Firstly, the EUO wood was applied for microwave hydrothermal pretreatment (MHTP), then the obtained wood precursor was activated with phosphoric acid in a traditional tube furnace (TTACs) or microwave tube furnace (MWACs), respectively. Then the structural properties, surface characteristics and electrochemical performance were evaluated. In detail, the hemicellulose contents of the EUO wood decreased significantly from 18.23 % to 1.23 % with MHTP temperature ranging from 150 °C to 190 °C. Moreover, the activated carbon generated via MHTP at 170 °C (MWAC170) possessed an excellent specific surface area (2138.3 m2/g) with a higher proportion of mesoporous structure (76.15 %) and the degree of graphitization (0.71 of ID/IG ratio) than those of samples by traditional pyrolysis method. MHTP significantly affected the component composition of the matrix, and the effect was amplificated in the microwave pyrolysis process. In this case, MWAC170 exhibited a high specific capacitance (346.86 F/g) and remarkable long-term cycling performance. The symmetric supercapacitor (SSC) offered a maximum energy density of 30.7 Wh/kg and power density of 4481.8 W/kg. Therefore, the activated carbon prepared by a two-step process combining MHTP and microwave pyrolysis could be an ideal electrode material for application in energy storage with excellent economic feasibility.

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.

Effects of Eucommia ulmoides Oliver leaves on gut microbiota and intestinal function during in vitro simulated digestion and fermentation

In this study, the effects of Eucommia ulmoides Oliver (EUO) leaves, both young and mature, on intestinal health were evaluated using in vitro simulated digestion and fermentation model. During the digestion phase, we observed that digestive enzymes gradually released polyphenols from the EUO leaves. Notably, the digestion products of EUO young leaves (EUO-YL) had higher polyphenol content and antioxidant activity compared to those of EUO mature leaves (EUO-ML). In the fermentation stage, the digestion products of EUO leaves effectively modulated the gut microbiota by increasing the relative abundances of short-chain fatty acids (SCFAs) producing bacteria, leading to higher SCFAs concentrations in the EUO leaf supplement groups. Additionally, the fermentation products of EUO leaves improved intestinal barrier function and decreased pro-inflammatory cytokine levels in Caco-2/THP1 co-cultures exposed to lipopolysaccharide (LPS). Our research supports the potential of EUO leaves as a functional food source for the prevention or treatment of related diseases.

Geniposide stimulates autophagy by activating the GLP-1R/AMPK/mTOR signaling in osteoarthritis chondrocytes

Osteoarthritis (OA) is a chronic joint disease characterized by cartilage degeneration. Autophagy is associated with chondrocyte homeostasis and exhibits a role in protecting against OA pathogenesis. Geniposide (GEN), an iridoid glycoside extracted from Eucommia ulmoides Oliv, acts as an activator of GLP-1R, which can stimulate autophagy. The AMPK/mTOR signaling pathway participates in the mediation of autophagy, and GLP-1R may act as an upstream factor of AMPK. However, whether GEN mediates the autophagic responses by activating the GLP-1R/AMPK/mTOR signaling pathway in OA chondrocytes is still unclear. In the current study, attenuated autophagy in MIA-induced rat OA models was observed, as shown by up-regulated expression of p62 and down-regulated expression of Beclin-1 and LC3-II/I. GEN stimulated autophagy and protected OA cartilage by up-regulating GLP-1R expression. In addition, GEN could enhance AMPK phosphorylation and down-regulate mTOR expression in IL-1β-treated C28/I2 cells. Inhibition of AMPK or activation of mTOR could reverse the stimulatory effects of GEN on autophagy. Furthermore, a GLP-1R inhibitor Exendin 9–39 could eliminate the chondroprotective effects of GEN by suppressing the AMPK/mTOR signaling pathway. Conclusively, Geniposide exhibits protective effects against osteoarthritis development by stimulating autophagy via activating the GLP-1R/AMPK/mTOR signaling pathway.

Eucommia ulmoides Oliver repairs the disorder of intestinal microflora caused by high starch in Micropterus salmoides and improves resistance to pathogens.

Eucommia ulmoides Oliver (EuO) is a natural medicine that can improve the composition of intestinal flora in fish, but more experiments and data are needed to support whether it can effectively improve the changes of intestinal flora and intestinal damage caused by high starch. This study examined the changes in intestinal structure as well as intestinal flora before and after the addition of EuO to high-starch diets and analyzed the effects of such changes on immune and digestive functions. The results showed that EuO reduces mortality during Nocardia seriolae attack and can reduce starch-induced intestinal inflammation. Eucommia ulmoides Oliver supplementation was able to alter the changes of intestinal flora in fatty acid degradation, bacterial chemotaxis, porphyrin metabolism and flagella assembly caused by high starch. By analyzing the abundance and correlation of bacterial communities, three bacterial communities that were significantly related to the intervention effect of EuO were screened. Further analysis revealed that EuO supplementation reduced the increase in abundance of Limnochordaceae, Nitrolancea, Lysinibacillus, and Hydrogenispora induced by high starch, which were negatively correlated with levels of the immunoreactive substance LZM in fish. This study reveals the regulatory effects of EuO on the intestinal flora of Micropterus salmoides fed on high starch diets, and provides a theoretical basis for reducing starch damage to fish in production.

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.

Cloning, Characterization, and Functional Analysis of EuTIL1, a Gene-Encoding Temperature-Induced Lipocalin in Eucommia ulmoides Oliv

Eucommia ulmoides Oliver is a unique tertiary relict tree species in China belonging to the Eucommia family and genus. It is a traditional and precious Chinese medicinal herb with anti-tumor, antibacterial, antioxidant, anti-inflammatory, and bidirectional blood pressure regulation effects. Eucommia ulmoides mainly grows in temperate regions of China, but due to its sensitivity to low-temperatures, it is difficult to introduce into new regions. To study the role of Eucommia ulmoides lipocalin in plants. This investigation was conducted utilizing gene cloning, bioinformatics analysis, quantitative real-time polymerase chain reaction (qRT-PCR), subcellular localization, and stable genetic transformation to transfer EuTIL1 into Nicotiana tabacum Xanthi. The wild-type transgenic vector and EuTIL1 tobacco were cold-treated, and the corresponding protective enzyme activity and cold-induced gene expression levels were measured to analyze the functions of the genes. In this study, the full-length of the temperature-induced lipocalin gene (EuTIL1) cDNA was cloned from the leaves of Eucommia ulmoides using the rapid amplification of cDNA ends (RACE) method. The sequence analysis showed that the full-length cDNA of EuTIL1 was 917 bp and encodes a protein of 188 aa residues, which is a member of the Lipocalin-2 family. Subcellular localization analysis revealed that EuTIL1 was found in the plasma membrane. The transgenic tobacco lines expressing EuTIL1 under the control of the CaMV 35S promoter had increased tolerance to cold compared to wild-type (WT) plants. The average water loss rate of EuTIL1 transgenic plants was 12.4%, the average conductivity at 24 h was 55.11%, and the malondialdehyde content at 24 h was significantly lower than that of wild-type plants. The maximum soluble sugar (SS) content, superoxide dismutase (SOD) activity, peroxidase (POD) activity, and catalase activity of EuTIL1 plants after low-temperature treatment were 22.03 mg/g, 726.87 U/g, 1283.94 U/g, and 356.84 U/g, respectively, which are significantly higher than those of the wild-type. Meanwhile, in the EuTIL1 transgenic tobacco plants, the expression of the NtDREB1, NtDREB2, NtDREB4, and NtCOR15a elevated under the low-temperature treatment condition. In conclusion, our study demonstrates that EuTIL1 is a gene involved in the cold-stress response and has the potential to enhance cold tolerance in plants, providing a potential molecular basis for the study of Eucommia ulmoides introduction and serving as a candidate gene for evaluating cold-tolerant plants.

Non-targeted lipidomics analysis of lipid distribution and variation of Eucommia ulmoides oliver by ultra-high performance liquid chromatography

Eucommia ulmoides Oliver (EU) is an important natural polymer source to relieve the serious shortage of natural rubber (NR), and the lipids of EU as potential non-rubber component have inherently a strong intramolecular connection to rubber biosynthesis. In this paper, Non-target lipidomics analysis platform based on ultra-high performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS) system was used to determine the lipids in EU. The orthogonal partial least square discriminant analysis (OPLS-DA) and a statistical analysis approach were used to compare the differences of lipids in EU. Total 6 categories, 26 classes and 785 species of lipids were identified in leaves, seed kernels, seed cases and barks of EU. The contents of EU lipids with time in female and male leaves were compared. The lipids contents in female EU leaves were more abundant than that in male EU leaves, and the content of most phospholipids in the female and male leaves both increased obviously after September. The content of lipids between seed cases and leaves were compared, and further screened for different glycerophospholipids between them. There were 53 phospholipids in the seed cases and fresh leaves samples of EU that had significantly different contents. Therefore, this study provides a theoretical basis for the identification of lipids in different EU organs and also helps further explore the relationship between lipids with rubber in EU.

Aucubin improves chronic unpredictable mild stress-induced depressive behavior in mice via the GR/NF-κB/NLRP3 axis

Eucommia ulmoides Oliv (EUO) is a traditional therapeutic drug that tonifies the liver and kidney and may improve depression. However, the mechanism of action of the main component, aucubin (AU), is unknown. To study the therapeutic effect of AU, we constructed a chronic unpredictable mild stress (CUMS) depression model in mice. Depression-like behaviors, pathological damage, hormonal changes, inflammation, intranuclear expression of glucocorticoidreceptor (GR), and hippocampal protein expression were assessed. Immunofluorescence staining of the hippocampus showed that CUMS decreased neuronal regeneration, and axons were observed to be reduced and broken. Intracellular GR expression decreased in the hippocampus and hypothalamus, and serum levels of stress hormones increased. Furthermore, molecular changes indicative of pyroptosis were observed. AU administration reversed these changes and significantly improved the depression-like behavior induced by CUMS. Our results suggested that AU improves depression by promoting the intranuclear expression of GR and inhibiting nuclear factor-kappa B-mediated inflammatory activation-driven cell pyroptosis.

Study on the substance basis of the efficacy of eucommiae cortex before and after salt processing for the treatment of kidney-yang deficiency syndrome based on the spectrum-effect relationship

Ethnopharmacological relevanceKidney-Yang deficiency syndrome (KYDS) is one of the common diseases of the elderly and closely related to the ageing of the body, it has a major impact on the quality of life of the patient. Eucommiae Cortex (EC) is the dried bark of Eucommia ulmoides Oliv. Which has the effect of tonifying the liver and kidneys, strengthening the muscles and bones. In Traditional Chinese Medicine clinics, EC is commonly used in the treatment of KYDS, but the material basis for the improvement of its efficacy in treating KYDS after salt processing remains unclear. Aim of the studyThis study aimed to find the main active ingredients that could improve the treatment of KYDS efficacy of EC after salt processing. Materials and methodsFirstly, the fingerprints of raw and salt-processed EC were established to determine the common components by using HPLC, and then an experimental study on the treatment of KYDS efficacy was carried out to compare the difference in the efficacy between raw and salt-processed EC. Thirdly, the spectrum-effect relationship of chemical components and pharmacodynamic indexes was established by using Grey Relational Analysis and Entropy Method. Finally, the network pharmacology and molecular docking technique was used to verify the kidney tonifying effect of the active ingredients of EC. ResultsAccording to the results of the analysis of hormonal index levels on the hypothalamic-pituitary-target gland axis and the extent of renal lesions, the therapeutic effect of EC on KYDS was mainly reflected in the regulation of the Adrenocorticotropic hormone, Corticosterone in the hypothalamic-pituitary-adrenal axis and Tri-iodothyronine, Tetra-iodothyronine in the hypothalamic-pituitary-thyroid axis, moreover the therapeutic effect of salt-processed EC was stronger than that of raw EC. The pharmacologically active ingredients that improved its treatment of KYDS efficacy after salt processing were peak 1 (geniposidic acid), peak 2 (chlorogenic acid), peak 5 (geniposide), peak 6 (genipin), peak 7 (pinoresinol diglucoside) and peak 11 (hyperoside). Meanwhile, the results of network pharmacology and molecular docking showed that the 6 active ingredients could exert kidney tonic effects through multiple signaling pathways by acting on core targets such as AKT1 and PTGS2. ConclusionAs far as we known, this was the first time to establish and compare the spectrum-effect relationship between raw and salt-processed EC, which laid the foundation for the pharmacokinetics studies of EC and provided a reference for future EC studies.