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In this study, the synergistic effect and weak gel mechanism of XG and Gleditsia sinensis polysaccharide (GSP) in different ratios were studied through the rheological properties, microstructure and molecular simulation based on density functional theory (DFT). The results of rheological properties showed that the mixtures formed a weak gel at the concentration of 0.5 % (w/v), with the synergistic impact peaking at a XG/GSP ratio of 3:7. Weak gels produced by XG and GSP had the intersection of G' and G" within the temperature sweep range, and the largest change in the G' slope at a XG/GSP ratio of 3:7. By calculating the interaction energy, it was found that the backbone of XG was more likely to interact with the backbone of GSP. Furthermore, the XG mainchain intersected with the backbone of GSP in a cross shape ("X" shape). As a result, this paper proposed a possible mechanism for the formation of the XG/GSP weak gel, with XG as the main chain and GSP as the grid point, and the main interaction type being hydrogen bonding, with the van der Waals force also involved. The results provide new insight for designing and producing physical gels with specific interactions in food industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Gleditsia fruits have been known as a valuable traditional Chinese herb for tens of centuries. Previous studies showed that the galactomannans are considered as one of the major bioactive components in Gleditsia fruits seeds (GSGs). Here, we systematically review the major studies of GSGs in recent years to promote their better understanding. The extraction methods of GSGs mainly include hot water extraction, microwave-assisted extraction, ultrasonic extraction, acid extraction, and alkali extraction. The analysis revealed that GGSs exhibited in the form of semi-flexible coils, and its molecular weight ranged from 0.018 × 10 3 to 2.778 × 10 3 KDa. GSGs are composed of various monosaccharide constituents such as mannose, galactose, glucose, and arabinose. In terms of pharmacological effects, GSGs exhibit excellent activity in antioxidation, hypoglycemic, hypolipidemic, anti-inflammation. Moreover, GSGs have excellent bioavailability, biocompatibility, and biodegradability, which make them used in food additives, food packaging, pharmaceutical field, industry and agriculture. Of cause, the shortcomings of the current research and the potential development and future research are also highlighted. We believe our work provides comprehensive knowledge and underpinnings for further research and development of GSGs., Competing Interests: Declaration of competing interest There is no conflict to declare., (Copyright © 2024. Published by Elsevier Ltd.)

The Gleditsia sinensis Lam. pods (GSP) are consistently discarded as waste after saponin extraction due to a lack of industrial or high-value utilization. Herein, the hemicelluloses were extracted from two varieties of GSP and subjected to comprehensive characterization. The molar mass of DMSO-soluble hemicelluloses (53.3-66.0 kDa) was higher compared to that of alkali-soluble ones (24.9-32.6 kDa). The presence of minimal acetyl substitution (3.85-4.49 %) on xylan was unequivocally confirmed. NMR spectroscopic analysis indicated that the hemicelluloses in GSP predominantly consist of a 1,4-β-ᴅ-Xyl backbone with arabinose substituents at O-3 and 4-O-methyl-α-ᴅ-GlcA substituents at O-2 of the xylose residues. p-Coumaric acid substitution also occurred on the 1,4-β-ᴅ-Xyl backbone. Hydrothermal treatment significantly reduced the hemicelluloses' relative molar mass and produced 7-10 % xylo-oligosaccharides. Furthermore, the moderately degraded hemicelluloses exhibited significantly enhanced biological activity. Finally, the incorporation of the moderately degraded hemicelluloses imparted the galactomannan film with exceptional antioxidant properties (81.1 % DPPH scavenging activity), while negligibly affecting its transparency. Our study's findings will contribute to a comprehensive understanding of the structural and biochemical properties of hemicellulose in waste G. sinensis pods, thereby facilitating their enhanced utilization in industrial applications., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Gleditsia sinensis , commonly known as Chinese Zaojiao, has important economic value and medicinal compounds in its fruits and thorns, making it widely cultivated artificially in China. However, the available literature on the impact of waterlogging on the growth of G. sinensis seedlings and the accumulation of metabolite compounds in its thorns is limited. To address this knowledge gap, G. sinensis seedlings were planted in soil supplemented with pindstrup substrate, which enhances the water-holding capacity of the soil. The analyses of morphological traits and nutrient elements in one-year-old G. sinensis seedlings grown naturally under ambient conditions and metabolite accumulation in its thorns were conducted. The results showed that the waterlogged soil significantly diminished the height, fresh weight, and dry weight of seedling roots and stems ( P  < 0.05). Furthermore, waterlogging hindered the uptake of iron (Fe) and manganese (Mn), as well as the transport of potassium (K). The identified metabolites within the thorns were categorized into 16 distinct groups. Relative to the control soil, fatty acids and derivatives were the most down-regulated metabolites in the waterlogged soil, accounting for 40.58% of the total metabolites, followed by lignans (38.71%), phenolic acids (34.48%), saccharides and alcohols (34.15%), steroids (16.67%), alkaloids (12.24%), flavonoids (9.28%), and glycerophospholipids (7.41%). Conversely, nucleotides and derivatives experienced the greatest up-regulation in the waterlogged soil, accounting for 50.00% of the total metabolites. In conclusion, waterlogging negatively impacted the growth of G. sinensis seedlings and inhibited the accumulation of metabolites. Hence, when considering the accumulation of secondary metabolites such as lignans and phenolic acids, appropriate management of soil moisture levels should be taken into account., Competing Interests: The authors declare there are no competing interests., (©2024 Luo et al.)

Medical and economic developments have allowed the human lifespan to extend and, as a result, the elderly population has increased worldwide. Osteoporosis is a common geriatric disease that has no symptoms and even a small impact can cause fractures in patients, leading to a serious deterioration in the quality of life. Osteoporosis treatment typically involves bisphosphonates and selective estrogen receptor modulators. However, these treatments are known to cause severe side effects, such as mandibular osteonecrosis and breast cancer, if used for an extended period of time. Therefore, it is essential to develop therapeutic agents from natural products that have fewer side effects. Gleditsiae fructus (GF) is a dried or immature fruit of Gleditsia sinensis Lam. and is composed of various triterpenoid saponins. The anti‑inflammatory effect of GF has been confirmed in various diseases, and since the anti‑inflammatory effect plays a major role in inhibiting osteoclast differentiation, GF was expected to be effective in osteoclast differentiation and menopausal osteoporosis; however, to the best of our knowledge, it has not yet been studied. Therefore, the present study was designed to examine the effect of GF on osteoclastogenesis and to investigate the mechanism underlying inhibition of osteoclast differentiation. The effects of GF on osteoclastogenesis were determined in vitro by tartrate‑resistant acid phosphatase (TRAP) staining, pit formation assays, filamentous actin (F‑actin) ring formation assays, western blotting and reverse transcription‑quantitative PCR analyses. Furthermore, the administration of GF to an animal model exhibiting menopausal osteoporosis allowed for the analysis of alterations in the bone microstructure of the femur using micro‑CT. Additionally, assessments of femoral tissue and serum were conducted. The present study revealed that the administration of GF resulted in a reduction in osteoclast levels, F‑actin rings, TRAP activity and pit area. Furthermore, GF showed a dose‑dependent suppression of nuclear factor of activated T‑cells cytoplasmic, c‑Fos and other osteoclastogenesis‑related markers.

Circular Gleditsia sinensis gum, Gleditsia microphylla gum, and tara gum are galactomannans (GMs) with similar mannose/galactose (M/G) molar ratios, which complicates the characterization of physicochemical properties using conventional methods. Herein, the hydrophobic interactions and critical aggregation concentrations (CACs) of the GMs were compared using a fluorescence probe technique, in which the I 1 /I 3 ratio of pyrene indicated polarity changes. With increasing GM concentration, the I 1 /I 3 ratio decreased slightly in dilute solutions below the CAC but decreased sharply in semidilute solutions above the CAC, indicating that the GMs formed hydrophobic domains. However, increases in temperature destroyed the hydrophobic microdomains and increased the CACs. Higher concentrations of salts (SO 4 2- , Cl - , SCN - , and Al 3+ ) promoted hydrophobic microdomain formation, and the CACs in Na 2 SO 4 and NaSCN solutions were lower than those in pure water. Hydrophobic microdomain formation also occurred upon Cu 2+ complexation. Although urea addition promoted hydrophobic microdomain formation in dilute solutions, the microdomains were destroyed in semidilute solutions and the CACs increased. The formation or destruction of hydrophobic microdomains depended on the molecular weight, M/G ratio and galactose distribution of GMs. Therefore, the fluorescent probe technique enables the characterization of hydrophobic interactions in GM solutions, which can provide valuable insight into molecular chain conformations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The self-healing property based on metal-ligand physical coordination is particularly interesting in bio-hydrogel science due to its allowance for multiple local healing events to process. As the most abundant renewable green resource in nature, Gleditsia sinensis galactomannan has great potential as a starting material for functional materials. In this study, the biocompatible Gleditsia sinensis galactomannan and cellulose were firstly chemically modified and then taken as the main constituent for constructing the metal-ligand coordination through an enzyme-regulated strategy. The hydrogel could quickly gelatinize in the surrounding environment, corresponding to the violent exothermic phenomenon, and exhibit extraordinary self-healing behavior. The molecular dynamics simulation of the hydrogel confirmed the more stable coordinated configuration from Fe(III)-chelates than Fe(II)-chelates. The morphology, mechanical property, antibacterial, and cytotoxicity of the prepared hydrogel were also studied. Our results indicated that galactomannan hydrogel based on the metal-ligand networks could balance the kinetic stability and intrinsic healability through the enzyme-induced route, which provide a new perspective in the field of biomaterial applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

A new flavonoid, (2''' E ,6''' S )-4''-(6-hydroxy-2,6-dimethylocta-2,7-dienoyl)-vitexin ( 1 ), and five known compounds ( 2 - 6 ) were isolated from the thorn of Gleditsia sinensis Lam. Their structures were determined by comprehensive and comparative spectroscopic analysis of NMR and MS data. The absolute configuration of the new compound was deduced by analysis of the experimental and calculated 13 C NMR data. The protective effects against lipopolysaccharide (LPS)-induced apoptosis in normal rat kidney tubule epithelioid (NRK 52e) cells of the isolated compounds were investigated in vitro , whose results showed that compound 1 showed significant protective effect with the EC 50 value of 3.0 μM.

Objective: Inflammation and oxidative stress are the major mechanisms implicated in lipopolysaccharide (LPS)-induced AKI. Spina Gleditsiae is a traditional Chinese anti-inflammatory medicine, from which a large number of flavonoids, such as 5-O-methyldihydroquercetin (GS1) and cilicicone B (GS2), were isolated in the present study. Here, we examined the reno-protective effects and potential underlying mechanisms of GS1 and GS2 in mice with LPS-induced AKI., Methods: We analyzed renal function; the serum metabolic profile, inflammatory cytokine levels, peripheral white blood cell count, renal cell apoptosis, renal oxidant and antioxidant levels, and renal expression levels of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), TIR-domain-containing adapter-inducing interferon-β (TRIF), nuclear factor-ĸB (NF-ĸB), interferon regulatory factor 3 (IRF3), NOD-, LRR- and pyrin domain-containing 3 (NLRP3, inflammasome), cleaved caspase-1, and interleukin 1 receptor type I (IL-1R1) in mice with LPS-induced AKI., Results: GS1 and GS2 improved renal function and significantly reduced the levels of inflammatory cytokines and oxidative stress markers. In addition, PCA score scatter plots suggest that the GS1 and GS2 groups were clustered with the control group, indicating that these compounds contributed to the recovery of mice with AKI toward the normal condition. Moreover, GS1 and GS2 inhibited the expression of TLR4, MyD88, TRIF, p-NF-ĸB, p-IRF3, NLRP3, cleaved caspase-1, and IL-1R1., Conclusion: The reno-protective effects of GS1 and GS2 are mediated via the MyD88/TRIF and NLRP3 pathways to reduce inflammation and oxidative stress through TLR4 signaling., Competing Interests: Conflict of interest There are no conflicts to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Gleditsia triacanthos polysaccharide, known as galactomannan, has not been exploited as a new functional material even though it possesses industrial potential in food and biomedicine. Galactomannans were recovered from the endosperm of seeds (15 weeks to 25 weeks after flowering) for deposition and maturation analysis. These galactomannans were characterized by using Nuclear magnetic resonance (NMR), X-ray diffraction (XRD), and monosaccharide composition analysis (particularly the mannose to galactose ratio) and molecular weight, solubility, and rheological measurements. The ratio of the three parts in mature seeds was as follows: endosperm (36.67%), hull (34.41%), and embryo (28.92%). The M/G ratio increased from 2.53 to 3.24 between 15 and 23 weeks and then decreased to 3.16 in 25 weeks, consistent with the trends of rheology and solubility. The molecular weight (1.28 × 10 6 g/mol) and intrinsic viscosity (882.53 mL/g) reached the maximum at 23 weeks and then decreased. Additionally, NMR and XRD showed that the M/G ratio did not change the basic chemical structure but caused slight changes in crystallinity. The purpose of the study was to reveal the changes in galactomannan structure, rheology, and solubility during G. triacanthos galactomannan deposition and maturation to facilitate exploration of its potential industrial applications., Competing Interests: Declaration of Competing Interest None., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Pods from a Gleditsia sinensis Lam. tree were collected and the galactomannan content and other properties were determined at their different developmental stages. In green and immature seed, galactomannan was substituted to a great extent with a mannose to galactose (M/G) ratio of 2.4 from crude polysaccharides. During late galactomannan deposition, it was substituted to a lower extent and this ratio increased rapidly, reaching a M/G ratio of 3.1. Average molecular weight (M w ) of the extracted polysaccharides first increased, reached the maximum (1.19 × 10 6 ) at 17 weeks after flowering (WAF), and then decreased. These changes might result from primary galactomannan biosynthesis and from galactose removal by α-galactosidase in the endosperm. The solubility of crude polysaccharides decreased with increased M/G ratio and maximum solubility was more than 89% that collected at 13 WAF. Rheological properties showed that apparent viscosity was largely influenced by the molecular weight and M/G ratio of galactomannans., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Numerous studies have focused on solvent extracts from locust trees (Gleditsia spp.), which contain diverse bioactive components including saponins, flavonoids, and alkaloids. However, because of the undefined nature of such phytochemicals, their clinical application as chemotherapeutic agents has often been limited., Purpose: This study aimed to evaluate the anti-oncogenic activity of triacanthine, an alkaloid obtained from Gleditsia triacanthos L., Study Design: The anti-oncogenicity of triacanthine in vitro was evaluated via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, cell-counting kit-8 assay (CCK-8 assay), flow cytometry, imunoblot, migration and invasion assays, zymography, and electrophoretic mobility shift assay in the human bladder carcinoma cell line EJ. The in vivo efficacy of triacanthine was evaluated via oral administration to EJ-xenografted BALB/c nude mice. To identify the side effects of triacanthine, cisplatin was also administered and an acute toxicity test was performed., Results: Triacanthine significantly inhibited EJ cell proliferation (IC 50 600 µM). Flow cytometry analysis revealed that cells were arrested in the G1 phase, and apoptotic cells accumulated in sub-G1 phase in a dose-dependent manner. Triacanthine inhibited the G1-S transition by deterring complex formation between cyclin-dependent kinases and cyclins, thereby up-regulating cell cycle inhibitors p21WAF1 and p27KIP1. In addition, triacanthine induced a caspase-dependent extrinsic pathway of apoptosis and autophagy. Early responsive kinases, extracellular signal-regulated kinase (ERK) and Janus kinase (JNK) were up-regulated by triacanthine. Triacanthine-mediated inhibition of the migratory and invasive potential of EJ cells was attributed to reduction of matrix metalloproteinase (MMP)-9 due to suppression of binding activities of the transcription factors activator protein (AP)-1, specificity protein (Sp)-1, and nuclear factor (NF)-κB. In an in vivo study, triacanthine significantly limited growth of xenografted tumors. Interestingly, while cisplatin resulted in significant weight loss after a 5-mg/kg dose, triacanthine did not cause weight loss, behavioral abnormalities, altered biochemical parameters, or tissue staining. A single oral dose acute-toxicity test (triacanthine 2,000 mg/kg) produced no adverse cytotoxic effects via blood biochemical tests and tissue-organ staining., Conclusion: To our knowledge, this is the first systematic evaluation of the anti-oncogenic activity of triacanthine. Therefore, we believe that our findings may guide the development of novel chemotherapeutic agents for bladder cancers., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

The thorns of Gleditsia sinensis have been historically used in Chinese medicine and are considered one of the fundamental therapeutic herbs. Its anticancer effects are currently being explored. Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and still requires the development of new drugs with higher efficiency. By using a rat HCC model implanted with cancerous Walker-256 cells, the therapeutic effects of G. sinensis extract (GSE) were assessed, as well as its regulatory effects on miRNAs. GSE significantly restored liver morphology and dramatically induced cell apoptosis in HCC rats. In addition, miR-21/181b/183 was upregulated in the HCC liver, and the elevation of these miRNAs could be alleviated by both GSE and sorafenib. PTEN/TIMP3/PDCD4 downregulation was consistent with the targets of miR-21/181b/183 in the HCC liver, and the alteration of these target genes was restored by both GSE and sorafenib. TIMP3 effects on MMP-2/9 expression were also determined. Our present findings indicate the potential of GSE in HCC treatment, and expand the understanding of miRNA-related mechanisms in the anticancer effects of GSE.

One new triterpenoid saponin ( 1 ), as well as six known ones ( 2-7 ), were isolated from the ethanol extract of the thorns of Gleditsia sinensis . Their structures were elucidated by extensive spectroscopic analysis in conjunction with chemical evidence. Cytotoxic activity of compounds 1-6 was evaluated against human breast cancer MCF 7 cells in vitro by the MTT method. Our results revealed moderate activities for compounds 1-6 with IC 50 values of 18.43, 30.47, 18.46, 10.02, 30.76, and 17.32 μM, respectively. Furthermore, compounds 1 , 3 , 4 , and 6 induced apoptosis in MCF 7 cell, with 1 and 6 causing late apoptosis of MCF 7 cells, while 3 and 4 acting oppositely.

Biofouling poses severe challenges to pearl oyster Pinctada imbricata culture in China, and controlling it is both labor- and capital-intensive. The antifouling properties of wax, and wax mixed with Chinese herbs, sprayed onto pearl oyster shell surfaces during peak biofouling seasons were evaluated. Pearl oysters coated with three wax treatments (plain wax, Chinaberry seed extract, Chinese honeylocust fruit extract) and a control (no treatment), were cultured in nets for up to 60 days. Mortality rate, fouling organism and pearl-oyster weights, and shell height are reported for individual oysters on each of six sampling dates. With the exception of oysters submerged for 12 days, all oysters were significantly affected by treatment type and submersion duration. Fouling weight increased more rapidly over time in the control-treatment oysters. Wax-based coatings deterred fouling-organism settlement on oysters for at least 2 months during the intensive fouling season, reducing mortality and not adversely effecting growth.

Gleditsia sinensis is widely used as a medicinal plant in Asia, especially in China. Triterpenes, alkaloids, and sterols were isolated from Gleditsia species. Among them, triterpenoid saponins are very important metabolites owing to their various pharmacological activities. However, the triterpenoid saponin biosynthesis pathway has not been well characterized. In the present study, we performed de novo transcriptome assembly for 14.3 Gbps of clean reads sequenced from nine tissues of G. sinensis. The results showed that 81,511 unique transcripts (unitranscripts) (47,855 unigenes) were constructed, of which 31,717 unigenes were annotated with Gene Ontology and EC numbers by Blast2GO against the NCBI-nr protein database. We also analyzed the metabolite contents in the same nine tissues by LS-MS/MS, and saponins including gleditsioside I were found in fruit at higher levels. Many of the genes with tissue-specific expression in fruit are involved in the flavonoid biosynthesis pathway, and many of those have UDP-glucosyltransferase (UGT) activity. We constructed a saponin biosynthesis pathway and identified two key enzyme families in the triterpenoid saponin biosynthesis pathway, cytochrome P450 and UDP-glucosyltransferase, that are encoded by 37 unigenes and 77 unigenes, respectively. CYP72A, CYP716A, and CYP88D, which are known as key enzymes for saponin biosynthesis, were also identified among the P450s. Our results provide insight into the secondary metabolite biosynthesis and serve as important resources for future research and cultivation of G. sinensis.

A new neolignan, 5-(3″-acetoxypropyl)-2-(4'-hydroxy-3'-methoxyphenyl)-7-methoxy-3-methylbenzofuran (1) along with nine analogues were isolated from the thorns of Gleditsia japonica var. delavayi by solvent extraction and repeated column chromatography. Their structures were elucidated by means of extensive spectroscopic analysis including 1D, 2D-NMR techniques and HR-ESIMS. All the isolates were reported for the first time from this species.

Waste paper has considerable potential as a raw material for lactic acid (LA) production due to high cellulose content, abundance and low cost. In this study, four kinds of waste papers were used for LA production through simultaneous saccharification and fermentation (SSF) by Streptococcus thermophilus. The SSF of office paper achieved the highest LA concentration (39.71 g/L), while the highest LA yield was observed for magazine (99.56%), followed by office paper (82.85%). High LA concentration is unfavorable to total LA conversion because of product inhibition. However, the addition of Gleditsia saponin (GS) could obtain both high yield and high concentration of LA at a low enzyme loading, indicating that product inhibition could be moderated. A lactic acid yield of 86.30% was obtained from office paper at an enzyme loading of 9 FPU/g-cellulose with GS, which was higher than that of without GS at a higher loading of 18 FPU/g-cellulose., (Copyright © 2018. Published by Elsevier Ltd.)

The Gleditsia sinensis Lam thorn (GST) is a classical traditional Chinese medical herb, which is of high medical and economic value. GST could be easily adulterated with branch of Rosa multiflora thunb (BRM) and Rosa rugosa thumb (BRR), because of their similar appearances and much lower cost for these adulterants. In this study Fourier transform near-infrared spectroscopy (FT-NIR) combined with chemical pattern recognition techniques was explored for the first time to discriminate and quantify of cheaper materials (BRM and BRR) in GST. The Savitzkye-Golay (SG) smoothing, vector normalization (VN), min max normalization (MMN), first derivative (1 st D) and second derivative (2nd D) methods were used to pre-process the raw FT-NIR spectra. Successive projections algorithm was adopted to select the characteristic variables and linear discriminate analysis (LDA), support vector machine (SVM), as while as back propagation neural network (BPNN) algorithms were applied to construct the identification models. Results showed that BPNN models performance best compared with LDA and SVM models for it could reach 100% accuracy for identifying authentic GST, and GST adulterated with BRM and BRR based on the spectral region of 6500-5500 cm -1 combined with 1 st D pre-processing. In addition, the BRM and BRR content in adulterated GST were determined by partial least squares (PLS) regression. The correlation coefficient of prediction (r p ), root mean square error of prediction (RMSEP) and bias for the prediction by PLS regression model were 0.9972, 1.969% and 0.3198 for BRM, 0.9972, 1.879% and 0.05408 for BRR, respectively. These results suggest that the combination of NIR spectroscopy and chemometric methods offers a simple, fast and reliable method for classification and quantification in the quality control of the tradition Chinese medicine herb of GST., (Copyright © 2018 Elsevier B.V. All rights reserved.)

To study morphological characteristics change of the development process and quercetin and polyphenols of accumulation dynamic of the spines of Gleditsia sinensis, measure and compare morphological indexes using Vernier caliper, ruler and balance, calibrate and analysis quercetin and polyphenols content using HPLC and colorimetric method. The spines of G. sinensis development process is divided into formation period (the beginning of August to the beginning of November), dormant period (the beginning of November to the end of March in the following year), germination period (the end of March to the middle of April), fast growth period (the middle of April to the middle of August), browning period (the middle of August to the end of August) and mature period (the beginning of September to the end of December). Formation period the spines primordium divides and forms the scale bubs; dormant period the scale bubs are in a dormant state; germination period the bubs scales fall off, spines primordium began to development; fast growth period rapid growth to maximum; browning period browning from the tip and browns until the whole becomes brown; mature period The early stage of maturity is full of luster, gradually the color deepened and the luster faded. The accumulation of quercetin was gradually decreasing after increasing. The total polyphenol accumulation was significantly decreased and then gradually increased, decreased finally. The content of quercetin was increased from 0.000 4%-0.002 6%, and the polyphenol content decreased from 0.761 9%-0.049 1% and then slowly increased to 0.286 9% in the fast growth period.The quercetin continuous increase to 0.004 3% and total polyphenol increased to 0.421 6% in the browning period. In the mature period, the quercetin content significantly decreased after reaching 0.009 6% in September, and the polyphenols content decreased after reaching 0.723 5% in October. Using principal component analysis results: September first, October 2nd, November 3rd. The morphological characteristics change of the development process and quercetin and polyphenols accumulation were determined. The development process is divided into six periods, the best harvest time is the early stage of mature period. Provide theoretical support for the utilization of the spines of G. sinensis and cultivation techniques of high yield., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Three new sulfated isoguanine alkaloid glycosides, designated as saikachinoside A monosulfate (1), saikachinoside A disulfate (2), and locustoside B disulfate (3), have been isolated from the pupal case of the wild bruchid seed beetle Bruchidius dorsalis (Chrysomelidae, Bruchinae) infesting the seed of Gleditsia japonica Miq. (Fabaceae). Their structures were determined by spectroscopic methods and the inhibitory activity of 2 and 3 against acid phosphatase was evaluated., (© 2018 Wiley-VHCA AG, Zurich, Switzerland.)

Gleditsiae Spina, the thorn of Gleditsia sinensis Lam., has been used as an anti-inflammatory, anti-tumor, and anti-bacterial traditional medicine for hundreds of years in China. This study used high-performance liquid chromatography and tandem mass spectrometry combined with chemometric methods to allow the fast and accurate identification and quantification of the flavonoids compounds in Gleditsiae Spina, and created reliable criteria for accurate identification of Gleditsiae Spina and its adulterants. This research provides good evidence for the classification and quality evaluation of Gleditsiae Spina. Firstly, eight flavonoids compounds were detected and identified on the basis of their mass spectra, fragment characteristics, and comparison with published data. Then the mass spectroscopic fragmentation pathways of these compounds were determined and, in addition rutin, isoquercitrin, and quercitrin were detected in Gleditsiae Spina for the first time. The quantification was performed on a triple quadrupole tandem mass spectrometer in multi-reaction monitoring mode, and the baseline separation of the eight bioactive flavonoids components was achieved within 13 min. Furthermore, the proposed method was successfully applied for simultaneous quantitative determination of the eight Gleditsiae Spina compounds and adulterants obtained from different sources in China. Then, we built a classification model which showed a high level of accuracy predicting 100% of the samples, correctly., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The water-soluble polysaccharides extracted from endosperm of Gleditsia japonica var. delavayi seeds (EGSP) were identified as galactomannan having the M/G ratio of 2.54-2.66 and a weight average molecular weight (Mw) of 1913 kDa. The molecular structure of EGSP was determined by periodate oxidation, Smith degradation, methylation, FTIR and NMR spectroscopy. The main chain is composed of β-1,4-d-mannopyranose and the branches composed of single α-1-d-galactopyranose. We had also established a model to speculate the fine structure of galactomannan molecules and given preliminary results. The I 2 -KI test indicated that there were many branches on the EGSP backbone and no starch in EGSP. The CD spectra and Congo red test showed EGSP was random coil conformations in solution and could form a small quantity of helical conformation under alkaline conditions. The microstructure of morphology was observed by OM, SEM and AFM. The results showed that the fibers composed of multiple microfibers formed a network construction by entangling with each other., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Three previously undescribed oleanane-type triterpenoidal saponins named caspicaosides L-N were isolated from the fruits of Gleditsia caspica Desf. The aglycons of these saponins were echinocystic acid, erythrodiol and 12-oleanene-3,28,30-triol. Caspicaoside L is a bisdesmosidic saponin acylated with two monoterpenic acids. It has a disaccharide moiety made up of glucose and arabinose attached to C-3 and pentasaccharide moiety linked to C-28 made up of one glucose, 2 xyloses, one inner rhamnose and one terminal rhamnose which was acylated with two identical monoterpenic acids. Caspicaoside M is a monodesmosidic saponin with a trisaccharide moiety at C-3 made up of glucose, xylose and arabinose, while caspicaoside N has a disaccharide moiety at C-3 made up of glucose and arabinose. Their structures were determined by extensive 1D and 2D (DQF-COSY, HSQC, TOCSY, 1 H- 13 C-HSQC-TOCSY, HMBC, ROESY, NOESY) NMR, HRESIMS analyses and chemical degradation. The cytotoxicity MTT-based assay showed that caspicaosides M, N and L, respectively, exhibited high cytotoxic activity with IC 50  ≤ 10 μM (72 h) at least against one of the three used cancer cell lines, MCF 7, A2780 and HT 29; and were 2-34 folds selective against the normal fibroblasts (MRC 5). All compounds also induced apoptosis and caused G 2 /M arrest in MCF 7 cells (24 h); thus showing pro-apoptotic properties., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Three previously undescribed isoguanine glycosides with an N 3 -prenyl group, designated locustoside B, saikachinoside B, and saikachinoside C, have been isolated from the seed of Gleditsia japonica Miquel (Fabaceae) along with two known compounds, locustoside A and saikachinoside A. Their structures were determined from spectroscopic data and X-ray crystallographic analysis. The inhibitory activity against acid phosphatase was evaluated., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Model systems formed by sodium caseinate (NaCAS) and espina corona gum (ECG) were studied. There was no evidence of attractive interactions between NaCAS and ECG macromolecules. Aqueous mixtures of NaCAS and ECG phase-separate segregatively over a wide range of concentrations. According to the images obtained by confocal laser scanning microscopy, NaCAS particles form larger protein aggregates when ECG is present in the system. An increase in the hydrodynamic diameter of NaCAS particles, as a result of ECG addition, was also observed by light scattering in diluted systems. A depletion-flocculation phenomenon, in which ECG is excluded from NaCAS surface, is proposed to occur in the concentrated mixed systems, resulting in NaCAS aggregation. ECG raises the viscosity of NaCAS dispersions without affecting the Newtonian flow behaviour of NaCAS. These results contribute to improve the knowledge of a barely-studied hydrocolloid which may be useful in the development of innovative food systems., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

One new flavanocoumarin (1), as well as six known flavonoids (2-7), was isolated from the ethyl acetate extract of the thorns of Gleditsia sinensis. The structures of these compounds were elucidated by extensive spectroscopic measurements and comparison with data reported in literatures. Cytotoxic activities of compounds 1-6 were evaluated against human liver cancer SK-hep-1 cells in vitro by the MTT method, with compound 1 displaying moderate activity (IC 50 of 62.53 μM). Furthermore, compound 1 could increase the number of apoptosis cells in a concentration-dependent manner.

The seeds of Gleditsia sinensis and Gleditsia microphylla , widespread in China, are an important source of galactomannans. G. sinensis gum (GSG) and G. microphylla gum (GMG) were purified and precipitated using different concentrations of ethanol and isopropanol. The GSG and GMG, precipitated in different stages, presented different characteristics, including polymer recovery, mannose/galactose ratio, chemical composition, molecular weight, and morphological appearance. The galactomannan recovery of GSG and GMG in 33.3% ethanol was 81.7% and 82.5%, respectively, while that in 28.8% isopropanol was 81.3% and 82.9%, respectively. To achieve similar precipitation efficiency, the amount of isopropanol should be lower than that of ethanol because of the lower dielectric constant of isopropanol (20 vs. 25 for ethanol). The precipitation behavior of galactomannans in polar organic solvents was dependent on the molecular structures and properties of the solvent. A higher mannose/galactose ratio and a higher molecular weight was obtained in a lower concentration of alcohols.

A comprehensive two-dimensional liquid chromatography coupled with mass spectrometry method was established to separate and characterize triterpenoid saponins in Gleditsia sinensis , the Chinese herbal medicine Zhu-Ya-Zao used for the treatment of apoplexy. The saponins were separated on an Agilent Zorbax Eclipse plus C 18 column (2.1 mm × 150 mm, 1.8 µm) for the first dimension, and a Poroshell 120 phenyl-hexyl column (3.0 mm × 50 mm, 2.7 µm) for the second dimension. Methanol and acetonitrile were used as the organic mobile phase for 1 D and 2 D, respectively. The theoretical peak capacity was 640, and the orthogonality was 57 %. Particularly, saponins with different numbers of monoterpene groups could be well separated on the second dimension. The structures were characterized by electrospray ionization tandem mass spectrometry in both positive and negative ion modes. In the MS/MS spectra, the relative abundances for [B α + Na] + and [Y 0α + Na] + were closely correlated with the number of sugar residues of the α -chain and β -chain, respectively, which facilitated the differentiation of isomers. Finally, a total of 72 saponins with molecular weights greater than 1500 Da were characterized. Among them, 49 compounds, including 2 acetylated saponins, were detected from G. sinensis for the first time., (Georg Thieme Verlag KG Stuttgart · New York.)

Ethnopharmacological Relevance: Zao-Jiao-Ci (ZJC), as the spine of Chinese Honey locust (Gleditsia sinensis Lam.), is traditionally used as Chinese medicine to reduce inflammation., Aim of the Study: The present study aimed to investigate an anti-inflammatory effect of ZJC aqueous extract both in vitro and in vivo, as well as its underlying mechanisms., Materials and Methods: Anti-inflammatory effect of ZJC aqueous extract was evaluated by using carrageenan-induced paw edema in rats. In addition, the inhibitory effects of ZJC on nitric oxide production, intracellular reactive oxygen species production, pro-inflammatory mediator expression and prostaglandin E 2 (PGE 2 ) production were determined by using LPS-activated RAW 264.7 cells. The anti-oxidant activity of ZJC was assessed using 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid assay., Results: ZJC aqueous extract showed significant suppressive effect on paw edema in rats at 100mg/kg. Moreover, ZJC aqueous extract decreased the expression of cyclooxygenase (COX)-2 and significantly decreased the PGE2, tumor necrosis factor-α, interleukin (IL)-1β and IL-6 production in LPS-activated macrophages in dose-dependent manners. ZJC aqueous extract inhibited the mRNA expression of these inflammatory cytokines as well. Furthermore, ZJC aqueous extract was found as an anti-oxidant and could inhibit ROS production in the LPS-induced cells., Conclusions: These findings show the potential of ZJC aqueous extract as a naturally occurring COX-2 inhibitor to reduce inflammation., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Chemical investigation of the anomalous fruits of Gleditsia sinensis led to the isolation and identification of a new triterpenoid saponin, 3-O-β-D-xylopyranosyl-(1 → 2)-α-L-arabinopyranosyl-(1 → 6)-β-D-glucopyranosyl oleanolic acid 28-O-β-D-xylopyranosyl-(1 → 4)-α-L-rhamnopyrano--syl-(1 → 4)-β-D-xylopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 3)-β-D-glucopyranosyl ester (1), along with other nine known compounds (2-10). All the isolates from this species were reported for the first time. The structure of Compound 1 was determined by a detailed analysis using various analytical techniques, including 1D and 2D NMR. In vitro antiproliferative activities of Compound 1 on MCF-7 and Hep-G2 tumor cell lines were evaluated. IC50 values against the two cell lines were 9.5 and 11.6 μM, respectively.

Gleditsia sinensis thorns (GST) have been used as a traditional medicine for carbuncles and skin diseases. The purpose of this study was to decide whether non-toxicological levels of water extract of GST (WEGST) are effective in inhibiting the progress of prostate cancer formation and to identify the target molecule involved in the WEGST-mediated inhibitory process of prostate cancer cell migration and in vivo tumor formation. Through the Boyden chamber migration assay, we found that non-toxic levels of WEGST could not attenuate the PC3 migration to the bottom area coated with serum but significantly inhibited PC3 cell migration to the collagen-coated bottom area. We also found that non-toxic levels of WEGST significantly attenuated collagen against adhesion. Interestingly, ectopic administration of WEGST could not affect the expression of α2β1 integrin, which is known as a receptor of collagen. However, when the PC3 cells adhered to a collagen-coated plate, the expression of α2 integrin but not that of β1 integrin was significantly inhibited by the administration of non-toxic levels of WEGST, leading to the inhibition of focal adhesion kinase (FAK) phosphorylation. Furthermore, oral administration of WEGST (25 mg/kg/day) significantly inhibited the size of a PC3 cell-xenografted tumor. Taken together, these results suggest a novel molecular mechanism for WEGST to inhibit prostate cancer progression at particular stages, such as collagen-mediated adhesion and migration, and it might provide further development for the therapeutic use of WEGST in the treatment of prostate cancer progression.

The apparent viscosity, molecular weight, and molecular weight distribution are important physical properties that determine the functional properties of galactomannan gum. Gleditsia sinensis gum (GSG) in semi-solid state was pressure cell treated over a range of temperature (30-110 °C) under nitrogen maintained at a pressure of 1.0-4.0 MPa. Physicochemical properties of GSG samples both before and after the pressure cell treatment were characterized. These include measurements of rheological properties by LVDV-III Ultra Rheometer, molecular weight and radius of gyration by light scattering, and changes in surface morphology by scanning electron microscopy. GSG had the highest apparent viscosity at a treatment temperature of 30 °C; further increase in temperature led to decrease in apparent viscosity. The apparent viscosity of GSG can be efficiently improved at room temperature and low pressure. The process of pressure cell treatment of GSG in semi-solid state could be industrialized for enhancement of rheological properties of galactomannan gum., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: The plants in the genus Gleditsia, mainly distributed in central and Southeast Asia and North and South America, have been used as local and traditional medicines in many regions, especially in China, for the treatment of measles, indigestion, whooping, smallpox, arthrolithiasis, constipation, diarrhea, hematochezia, dysentery, carbuncle, etc. This present paper systemically reviews the miscellaneous information surrounding its traditional use, phytochemistry and pharmacology to provide opportunities and recommendations for the future research., Materials and Methods: The scientific literatures were systematically searched from scientific databases (PubMed, Scopus, Elsevier, SpringerLink, SciFinder, Google Scholar and others). In addition, the ethnopharmacological information on this genus was mainly acquired from Chinese and Korean herbal classics, and library catalogs., Results: More than 60 compounds including triterpenes, sterols, flavonoids, alkaloids, phenolics and their derivatives were isolated from Gleditsia japonica Miq., Gleditsia sinensis Lam., Gleditsia caspica Desf. and Gleditsia triacanthos L. Among these compounds, triterpenoid saponins were the main constituents of Gleditsia species. Moreover, the crude extracts and purified molecules were tested, revealing diverse biological activities such as anti-tumor, anti-inflammatory, anti-allergic, anti-hyperlipidemic, analgesic, antimutagenic, antioxidant, anti-HIV, antibacterial, antifungal activities, etc. Among these biological studies, the possible mechanisms of antitumor action are stressed in this review, and these include causing cytotoxicity to cancer cells, inhibition of proliferation of cancer cells by affecting their growth, regeneration and apoptosis, inhibition of basic fibroblast growth factor (bFGF) and nitric oxide (NO), modulation of the oncogenic expression and telomerase activity results, inhibition of the expression of pro-angiogenic proteins, as well as down-regulation of intra/extracellular proangiogenic modulators, etc., Conclusions: On the basis of preliminary research on Gleditsia genus it could be stated that saponins investigations may be more promising in future. Although 32 compounds of 67 identified compounds were saponins, modern pharmacological research on saponins were not a priority in Gleditsia species. Therefore, more bioactive experiments and in-depth mechanisms of action are required for elucidating their roles in physiological systems. Moreover, the present review also highlights that analgesic, anti-tumor and anti-HIV activities should have priority in saponins research. Additionally, it is imperative to explore more structure-activity relationships and possible synergistic actions of triterpenoid saponins for revaluating their pharmacological activities., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

The fruits of Gleditsia species (Fabaceae) have been known in traditional medicine as a saponin-rich herbal medicine. The present study aimed to investigate the effects of the total methanolic extract of Gleditsia caspica (MEGC) and its saponin-containing fractions (SFGC) on hyperglycemia in streptozotocin (STZ)-induced diabetic rats. A single intraperitoneal injection of STZ (55 mg/kg body weight) was used to induce hyperglycemia in male albino rats. MEGC (15, 30 and 60 mg/kg, p.o.) and SFGC (15, 30 and 60 mg/kg, p.o.) were administered to the diabetic rats daily for 14 days. The anti-diabetic drug gliclazide (10 mg/kg, p.o.) was used as a positive control. Blood samples were collected from overnight fasted rats for the evaluation of the antihyperglycemic, antihyperlipidemic and antioxidant activities. The levels of glucose, triglycerides (TG), cholesterol (TC) and malondialdehyde (MDA) were increased significantly, whereas the levels of α-amylase, insulin and reduced glutathione (GSH) were decreased in the experimental diabetic rats. Pancreas and liver of the diabetic rats exhibited significant changes in the histopathology, morphology and DNA content. Administration of MEGC or SFGC led to a decrease in the levels of glucose, TG, TC and MDA. In addition, the levels of α-amylase, insulin and GSH were increased in MEGC and SFGC treated diabetic rats. Also, the histopathological and morphological changes, as well the changes in DNA were significantly reversed by the extracts. Thus, MEGC and SFGC exhibited potent hypoglycemic and hypolipidemic activities in STZ- induced diabetic rats.

Gleditsia sinensis thorns have traditionally been used to treat edema and carbuncles and drain abscesses. In the present study, a simultaneous analysis of four flavonoids [(+)‑catechin, (‑)‑epicatechin, eriodictyol and quercetin] and two phenolic compounds (caffeic acid and ethyl gallate), obtained from a 70% ethanol extract of G. sinensis, was performed using high‑performance liquid chromatography‑photodiode array techniques. In addition, the inhibitory activities of the solvent fractions from a G. sinensis extract and its major constituents on the lipopolysaccharide‑stimulated production of inflammatory mediators by macrophage RAW 264.7 cells and the tumor necrosis factor (TNF)‑α and interferon (IFN)‑γ (TI)‑stimulated production of chemokines by HaCaT keratinocyte cells were investigated. The established analytical method showed high linearity, with a correlation coefficient of ≥0.9998. The limits of detection and quantification of the six compounds were 0.037‑0.425 and 0.124‑1.418 µg/ml, respectively. The ethyl acetate fraction inhibited nitric oxide and prostaglandin E2 production in RAW 264.7 cells and the production of thymus‑ and activation‑regulated chemokine (TARC) in HaCaT cells more than did the other fractions. Furthermore, the six compounds reduced the production of TARC, macrophage‑derived chemokine and regulated on activation normal T‑cell expressed and secreted in TI‑stimulated HaCaT cells; in particular, ethyl gallate and quercetin exhibited a significant dose‑dependent inhibition. Further elucidation of the signaling pathways involved in the T‑helper cell 2 chemokine inhibition by G. sinensis is necessary to facilitate the design of therapeutic agents for the inflammatory response.

Angiogenesis is the process of new vessel formation from pre-existing blood vasculature and is critical for continuous tumor growth. We previously reported that an ethanolic extract of Gleditsia sinensis thorns (EEGS) and its active constituent, cytochalasin H, have anti-angiogenic activity in vitro and in vivo via suppression of endothelial cell functions. In the present study, EEGS and cytochalasin H were observed to efficiently inhibit tumor growth in an in ovo xenograft model without significant toxicity. We repeatedly observed the anti-tumor and anti-metastatic effects of EEGS in representative animal models. These results suggest that EEGS and its active constituent, cytochalasin H, are potential candidates for the development of anti-angiogenic cancer drugs.

Three triterpenoidal saponins were isolated from the saponin fraction derived from a Gleditsia caspica Desf. methanolic fruit extract. The isolated saponins were identified as gleditsiosides B, C, and Q based on spectral data. The saponin-containing fraction was evaluated in vivo for genotoxic and antigenotoxic activities. The fraction caused no DNA damage in Swiss albino male mice treated with a dose of 45 mg/kg body weight for 24 h, although it significantly inhibited the number of chromosomal aberrations induced by cyclophosphamide (CP) in bone marrow and germ cells when applied before or after CP administration. The inhibitory indices in chromosomal aberrations were 59% and 41% for bone marrow and 48% and 43% for germ cells, respectively. In addition, the saponin fraction was found to reduce the viability of the human tumor cell line MCF-7 in a dose-dependent manner with an extrapolated IC50 value in the range of 220 μg/mL.

Background: The fruit hull of Gleditsia sinensis (FGS) used in traditional Asian medicine was reported to have a preventive effect on lung inflammation in an acute lung injury (ALI) mouse model. Here, we explored FGS as a possible therapeutics against inflammatory lung diseases including ALI, and examined an underlying mechanism for the effect of FGS., Methods: The decoction of FGS in water was prepared and fingerprinted. Mice received an intra-tracheal (i.t.) FGS 2 h after an intra-peritoneal (i.p.) injection of lipopolysaccharide (LPS). The effect of FGS on lung inflammation was determined by chest imaging of NF-κB reporter mice, counting inflammatory cells in bronchoalveolar lavage fluid, analyzing lung histology, and performing semi-quantitative RT-PCR analysis of lung tissue. Impact of Nrf2 on FGS effect was assessed by comparing Nrf2 knockout (KO) and wild type (WT) mice that were treated similarly., Results: Bioluminescence from the chest of the reporter mice was progressively increased to a peak at 16 h after an i.p. LPS treatment. FGS treatment 2 h after LPS reduced the bioluminescence and the expression of pro-inflammatory cytokine genes in the lung. While suppressing the infiltration of inflammatory cells to the lungs of WT mice, FGS post-treatment failed to reduce lung inflammation in Nrf2 KO mice. FGS activated Nrf2 and induced Nrf2-dependent gene expression in mouse lung., Conclusions: FGS post-treatment suppressed lung inflammation in an LPS-induced ALI mouse model, which was mediated at least in part by Nrf2. Our results suggest a therapeutic potential of FGS on inflammatory lung diseases.

Aims: Our previous studies revealed that echinocystic acid (EA) showed obvious attenuation of atherosclerosis in rabbits fed a high-fat diet. However, the underlying mechanisms remain to be elucidated. Considering the importance of endothelial progenitor cells (EPCs) in atherosclerosis, we hypothesise that EPCs may be one of the targets for the anti-atherosclerotic potential of EA., Main Methods: After in vitro cultivation, EPCs were exposed to 100 μg/mL of oxidised low-density lipoprotein (oxLDL) and incubated with or without EA (5 and 20 μM) for 48 h. An additional two groups of EPCs (oxLDL+20 μM EA) were pre-treated with either wortmannin, an inhibitor of the phosphoinositide 3-kinase (PI3K) pathway, or nitro-l-arginine methyl ester (l-NAME), an endothelial nitric oxide synthase (eNOS)-specific inhibitor. Assessment of EPC apoptosis, adhesion, migration, and nitric oxide (NO) release was performed using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) staining, cell counting, caspase-3 activity assay, transwell chamber assay, and Griess reagent, respectively. The protein expression of protein kinase B (Akt) and eNOS was detected using Western blot., Key Findings: Treatment of EPCs with oxLDL induced significant apoptosis and impaired adhesion, migration, and NO production. The deleterious effects of oxLDL on EPCs were attenuated by EA. However, when EPCs were pre-treated with wortmannin or l-NAME, the effects of EA were abrogated. Additionally, oxLDL significantly down-regulated eNOS protein expression as well as repression of eNOS and Akt phosphorylation., Significance: The inhibitory effect of oxLDL on Akt/eNOS phosphorylation was attenuated by EA. Taken together, the results indicate that EA protects EPCs from damage caused by oxLDL via the Akt/eNOS pathway., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The thorns of Gleditsia sinensis have been used to prevent or treat numerous diseases. The present study aimed to investigate the molecular mechanism of the ethanol extract of Gleditsia sinensis thorns (EEGS) on platelet-derived growth factor (PDGF)‑treated vascular smooth muscle cells (VSMCs). EEGS treatment was found to inhibit DNA synthesis in PDG\F-treated VSMCs in a dose-dependent manner, without cell toxicity. These inhibitory effects were associated with G1-phase cell-cycle arrest, which was caused by the decreased expression of cyclins and cyclin-dependent kinases (CDKs) and the upregulation of p27KIP1 expression in PDGF-stimulated VSMCs. Among the pathways examined, EEGS treatment was observed to only inhibit the PDGF‑induced phosphorylation of Akt. In addition, EEGS treatment suppressed the migration and invasion of VSMCs in the presence of PDGF as determined by wound-healing and Matrigel™ invasion assays. Furthermore, zymographic and western blot analyses revealed that EEGS treatment suppressed matrix metalloproteinase (MMP)-9 expression in PDGF‑treated VSMCs, which was attributed to a reduction in the binding activities of nuclear factor κ-light-chain-enhancer of activated B cells (NF‑κB), activator protein (AP)‑1 and specificity protein (Sp)‑1. These results demonstrate that EEGS induces p27KIP1‑mediated G1-phase cell-cycle arrest, reduces Akt phosphorylation and prevents MMP‑9 expression by decreasing the binding activities of NF‑κB, AP‑1 and Sp‑1 in PDGF-treated VSMCs, thus resulting in growth inhibition and the suppression of migration and invasion. These results may suggest a novel perspective for the use of EEGS in the treatment and prevention of vascular proliferative diseases.

Background: The anomalous fruits of Gleditsia sinensis Lam. (Leguminosae), a crude drug in China, have long been used in traditional Chinese medicine for the treatment of various diseases. The saponin fraction isolated from the fruits (SFGS) is considered as the active component for the antitumor activity of this crude drug., Objectives: The present study was performed to investigate the anti-angiogenesis activities and active constituents of SFGS., Methods: Human umbilical vein endothelial cells (HUVECs) were treated with SFGS in the presence or absence of basic fibroblast growth factor (bFGF) in vitro. The proliferation, migration, and tube formation were studied by MTT, Transwell, and 2D Matrigel assays, respectively. The cell cycle and apoptosis were analyzed by flow cytometry. Enzyme-linked immunosorbent assay for protein expression of vascular endothelial growth factor (VEGF) and western blot analysis for caspase-3, caspase-8, and caspase-9 as well as Fas were performed. In addition, the effects of 13 saponin compounds isolated from SFGS on the tube formation of HUVECs were screened, and the structure-activity relationships were discussed., Results: SFGS, at concentrations (1, 3, and 10 µg/mL) without significant cytotoxicity on endothelial cells, significantly inhibited the proliferation, migration, and tube formation of HUVECs induced by bFGF (10 ng/mL). It moderately arrested the cell cycle to G1 phase but greatly induced cell apoptosis and increased the expressions of caspases-3, caspase-8, and Fas but not caspase-9 in HUVECs. Moreover, SFGS did not affect the bFGF-induced autosecretion of VEGF from endothelial cells. Among the 13 saponin compounds tested, gleditsiosides B, I, J, O, and Q showed inhibition of the tube formation at a concentration of 3 µM, and only gleditsioside B exerted significant inhibition at 1 µM., Conclusion: SFGS is substantially able to prevent angiogenesis by interfering with multiple steps. The findings provide a new explanation for the antitumor effects of G sinensis fruits. Gleditsiosides B, I, J, O, and Q are probably the main active constituents of SFGS., (© The Author(s) 2012.)

Seven bisdesmosidic triterpenoid saponins named caspicaosides E-K, were isolated from the methanolic fruit extract of Gleditsia caspica Desf. Their structures were determined by 1D and 2D NMR spectroscopy as well as high resolution mass spectrometry and acid hydrolysis. The saponins comprised echinocystic acid or oleanolic acid as the aglycone and saccharide moieties at C-3 and C-28. Like most Gleditsia saponins, the oligosaccharide moiety at C-3 was identified as β-D-xylopyranosyl-(1→2)-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranosyl. The common oligosaccharide moiety linked to C-28 was determined as β-D-xylopyranosyl-(1→3)-β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl with the presence of additional β-D-galactopyranose unit and/or another α-L-rhamnopyranose moiety. The C-28 saccharide moiety was acylated with a monoterpenic acid unit or a monoterpenic acid linked to a monoterpene-arabinoside unit at the ester-β-D-glucopyranose C-6. Esterification of C-2 and C-3 hydroxyl groups of the terminal α-L-rhamnopyranose unit with a monoterpenic acid and epoxy-monoterpenic acid units, respectively, or with two identical monoterpenic acid units, was also shown. The acylated saponins caspicaosides G-K were assayed for their in vitro cytotoxicities against the three cell lines HCT116, HepG2 and MCF7. The tested saponins showed moderate to strong activities., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Angiogenesis, the process of new vessel formation from the pre-existing blood vasculature, is critical for continuous tumor growth and is considered to be a validated antitumor target. The results of our previous study demonstrate the anti-angiogenic potential of an extract of Gleditsia sinensis thorns, which has been traditionally used in Korean medicine to remedy diverse diseases, including tumors. In the present study, we attempted to identify the active anti-angiogenic constituents of the ethanol extract of G. sinensis thorns (EEGS). By virtue of in vitro activity-guided fractionation using human umbilical vein endothelial cells (HUVEC) primary endothelial cells, chromatographic separation, and NMR spectral analyses, we isolated and identified the potent active constituent, cytochalasin H, a biologically active secondary metabolite of fungi. This unexpected active constituent may have originated from the endophytic fungi, Chaetomium globosum, which naturally populate G. sinensis, the identity of which was determined by analysis of fungal community. Cytochalasin H isolated from the EEGS showed in vitro anti-angiogenic activities such as suppressed cell growth and mobility in HUVEC, and inhibited the pro-angiogenic protein-induced formation of new blood vessels in vivo. The anti-angiogenic effect of cytochalasin H was in part due to reduced expression of pro-angiogenic factor, such as endothelin-1. This is the first report regarding the isolation and identification of cytochalasin H, as an active anti-angiogenic constituent of G. sinensis thorns.

Gleditsiae Fructus Abnormalis and Gleditsiae Sinensis Fructus are obtained from different developmental stages of fruits from Gleditsia sinensis Lam. (Leguminosae). The possible interchangeable usage of the two fruits, however, has long been very controversial. Here, high performance liquid chromatography coupled with diode array detection was developed to explore their chemical fingerprinting profiles. Besides, the amounts of aglycones of saponin compounds, echinocystic acid and oleanolic acid in both fruits were quantified. The results indicated that there was no significant difference in the content of aglycones from the two types of fruits. However, their chromatographic fingerprints showed distinct characteristics. Therefore, the interchangeable application of these fruits has to be taken with a specific precaution.

The physico-chemical properties and hydration kinetics of Gleditsia sinensis gum were investigated to evaluate its temperature dependence. The increase of temperature resulted in improved solubility of G. sinensis gum, and the dissolved galactomannan showed decreased degree of galactose substitution (DSGal) and increased molecular weight (p<0.05, ANOVA). The solubility of G. sinensis galactomannan at 25, 45, 60 and 80 °C was 73.4, 83.8, 98.1, and 99.4%, respectively. Weibull and Logarithmic models were well fitted (R(2)>0.96), and the hydration index t0.8 at different temperatures varied in the range of 51-302 min. It was found that galactomannan with low DSGal and high molecular weight exhibited slow hydration rate and poor solubility., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The thorns of Gleditsia sinensis are a traditional Oriental medicine used for the treatment of swelling, suppuration, carbuncle and skin diseases. In the present study, we identified a novel molecular mechanism by which an ethanol extract of Gleditsia sinensis thorns (EEGS) inhibits the growth of the SNU-5 human gastric cancer cell line. EEGS treatment inhibited cell growth and was associated with G1 phase cell cycle arrest at a concentration of 400 µg/ml (IC50) in SNU-5 cells. Treatment with EEGS also stimulated p21WAF1 expression, which significantly decreased the expression of cyclins and cyclin-dependent kinases (CDKs). Further study suggested that p38 MAP kinase pathways may be involved in the inhibition of cell proliferation through p21WAF1‑dependent G1 phase cell cycle arrest in EEGS-treated cells. In addition, NF-κB and AP-1 transcription factor binding sites were identified as the cis-elements for tumor necrosis factor-α (TNF-α)-induced matrix metalloproteinase-9 (MMP-9) expression in SNU-5 cells, as determined by gel-shift assay. Treatment of cells with EEGS suppressed MMP-9 expression induced by TNF-α via a decrease in the binding activity of both NF-κB and AP-1 motifs. These data demonstrate that EEGS-mediated inhibition of cell growth appears to involve the activation of p38 MAP kinase, subsequently leading to the induction of p21WAF1 and the downregulation of cyclin D1/CDK4 and cyclin E/CDK2 complexes. Moreover, EEGS strongly inhibited TNF-α-induced MMP-9 expression by impeding the DNA binding activity of NF-κB and AP-1. Overall, these results provide a potential mechanism for EEGS in the treatment of gastric cancer.

Enzymatic hydrolysis of Gleditsia sinensis gum was performed to produce manno-oligosaccharides having functional applications as dietary fiber and prebiotics. The optimum hydrolysis conditions, including enzyme loading, temperature and time, from response surface methodology were 8.1 U/g, 57.4 °C and 34.1 h, respectively. The yield of DP 1-5 oligosaccharides was 75.9% (29.1 g/L). The Michaelis-Menten kinetics and molecular weight distribution were determined. The obtained oligosaccharides were further separated by HPLC and SEC, and the galactose distribution of G. sinensis gum was elucidated. Results indicated that G. sinensis gum has potential to produce value-added oligosaccharides in food industries., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A high performance liquid chromatography method was developed for simultaneous quantification of 29 ingredients occurring as a very complex mixture in the gleditsia fruits. The analysis was performed on an ODS-2 Hypersil column (250 mm × 4.6 mm, 5 μm) with a binary gradient mobile phase of water and acetonitrile both containing 0.1% acetic acid. The method was validated in terms of linearity, sensitivity, stability, precision, and accuracy. It was found that this method had linearity with R² at 0.99889-0.99997 in the test range of 1.0-24.0 μg. The limit of detection (LOD) and limit of quantification (LOQ) for 16 tested reference saponins were 0.24-0.39 μg and 1.0-1.2 μg, respectively. The relative standard deviations (RSDs %) for intra-day and inter-day repeatability were not more than 3.11% and 4.02%, respectively. The analyzed samples were stable for at least 48 h. The spike recoveries for eight analyzed saponins were 99.68-102.17%. The established HPLC analytic method was successfully used to determine the concentrations of 29 compounds including 19 gleditsia saponins and ten unidentified ingredients in eight commercial gleditsia fruits from different sources. The results from this study suggested that this newly developed HPLC method could be used for qualitative and quantitative analysis of the saponins in the gleditsia fruits and the gleditsia extracts that used for animal study and other purpose., (Copyright © 2012 Elsevier B.V. All rights reserved.)