Ligustrum Lucidum Ait Research Articles

Nuezhenide of the fruits of Nuzhenzi (Ligustrum lucidum Ait.) is a functional analog of ghrelin

Ethnopharmacological relevanceThe dried fruit of Ligustrum lucidum Ait. (FLL), known as Nuzhenzi, is traditionally recognized for its anti-aging properties in Chinese medicine. Nuezhenide, a water-soluble secoiridoid present in FLL, has demonstrated various pharmacological activities including neuroprotection, enhancement of learning and memory, antiosteoporotic, and antiviral activities. These therapeutic benefits align with the anti-aging effects attributed to ghrelin, particularly in the modulation of growth hormone secretagogue receptor type 1a (GHSR-1a) signaling. Aim of the workThis study aimed to investigate the potential of FLL extracts, particularly its major compound nuezhenide, as agonists of GHSR-1a, a receptor implicated in anti-aging mechanisms, utilizing a stable GHSR-1a-expressing cell line. Materials and methodsHEK293T cells expressing GHSR-1a-mCherry were used to assess the effects of FLL extract and its major compound, nuezhenide, on cell viability and ERK1/2 signaling. Molecular docking simulations predicted the interaction between nuezhenide and the GHSR-1a binding pocket. The impact of nuezhenide on ERK1/2 phosphorylation was evaluated, along with the involvement of phospholipase C and calcium signaling in this process. ResultsMolecular docking simulations indicated that nuezhenide could interact with the GHSR-1a receptor, similar to teaghrelin, another known ghrelin analog. Experimental data showed that FLL extracts and nuezhenide enhanced cell viability and ERK1/2 activation in GHSR1a-mCherry HEK293T cells. The effect was specifically mediated by GHSR-1a, as confirmed by SP-analog treatment. Further analysis revealed that nuezhenide-induced ERK1/2 activation is likely mediated through a phospholipase C-dependent pathway involving intracellular calcium release. ConclusionThis study demonstrated for the first time that nuezhenide acts as a putative GHSR-1a agonist, promoting cell proliferation and activating ERK1/2 signaling via phospholipase C and calcium pathways. These findings support the traditional use of FLL as an anti-aging herbal remedy and suggest that nuezhenide could be developed as a therapeutic agent targeting GHSR-1a-mediated pathways.

Synthesis, Anti-Oomycete and Anti-Fungal Activities of Anhydride Derivatives of Oleanolic Acid.

Oleanolic acid is a pentacyclic triterpenoid extracted and isolated from the fruit of plants in the Ligustrum lucidum Ait. in the family Oleaceae. To discover biorational natural product-based pesticides, a series of oleanolic acid derivatives containing anhydride active skeletons were prepared by ingeniously introducing an active acyloxy group at its C-28 carboxyl position, and their structures were well characterized by 1H NMR, 13C NMR, HRMS, and m.p.. The stereochemical configuration of compound 8e was confirmed using single-crystal X-ray diffraction. Furthermore, bioactivities of these compounds as anti-oomycete and anti-fungal agents against two serious agricultural pests, Phytophthora capsici and Fusarium graminearum we assessed. Amongst evaluated compounds, 1) Compounds 8h and 8j displayed significant anti-oomycete against P. capsici, with EC50 values of 54.73 and 65.15 mg/L, respectively. 2) The target compounds have obvious selectivity, and their anti-oomycete activity is significantly better than their anti-fungal activity. 3) Interestingly, there are significant differences in the structure-activity relationship of different substituents or the same substituent at different positions anti-oomycete and anti-fungal against P. capsici and F. graminearum, respectively. The study provides an idea for further exploring the bioactivities of 28-acyloxyoleanolic acid derivatives, and develops the application of 28-acyloxyoleanolic acid derivatives containing anhydride in agriculture.

Impact of different processing methods of Ligustrum lucidum Ait. on kidney-yin deficiency: a study based on pharmacodynamics and metabolomics research.

This study aimed to explore the pharmacodynamics and mechanisms of different processing methods of Ligustrum lucidum Ait. (LLA) in addressing kidney-yin deficiency (KYD). Forty-eight Sprague-Dawley rats were divided into eight groups based on their weight. The KYD model was established by intragastric administration of levothyroxine sodium. Each group was administered the corresponding treatment for 15 consecutive days. The general condition of the rats during the treatment period was observed. In addition, the levels of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), and the ratio of cAMP to cGMP in the serum of rats from different groups were measured. Serum samples were analyzed using the ultra-performance liquid chromatography (UPLC)-Orbitrap Fusion MS technique for metabolomics analysis. Compared with the model group, the general condition of the rats in the wine-steamed L. lucidum group (WL) and salt-steamed L. lucidum group (SSL) groups showed significant improvement. The serum levels of cAMP, cGMP, and the cAMP-to-cGMP ratio tended to return to normal. Metabolic analysis identified 38 relevant biomarkers and revealed 3 major metabolic pathways: phenylalanine, tyrosine, and tryptophan biosynthesis; phenylalanine metabolism; and sphingolipid metabolism. The different processing methods of LLA demonstrated therapeutic effects on KYD in rats, likely related to the restoration of disturbed metabolism by adjusting the levels of endogenous metabolites in the kidney. The SSL demonstrated significantly superior effects compared with the other four types of LLA processed products.

Specneuzhenide improves bleomycin-induced pulmonary fibrosis in mice via AMPK-dependent reduction of PD-L1

BackgroundPulmonary fibrosis (PF) is an escalating global health issue, characterized by rising rates of morbidity and mortality annually. Consequently, further investigation of potential damage mechanisms and potential preventive strategies for PF are warranted. Specnuezhenide (SPN), a prominent secoiridoid compound derived from Ligustrum lucidum Ait, exhibits anti-inflammatory and anti-oxidative capacities, indicating the potential therapeutic actions on PF. However, the underlying mechanisms of SPN on PF remain unclear. PurposeThis work was aimed at investigating the protective actions of SPN on PF and the potential mechanism. MethodsIn vivo, mice were administrated with bleomycin (BLM) to establish PF model. PF mice were treated with SPN (45/90 mg/kg) by gavage. In vitro, we employed TGF-β1 (10 ng/mL)-induced MLE-12 and PLFs cells, which then were treated with SPN (5, 10, 20 µM). DARTS assay, biofilm interference experiment and molecular docking were performed to investigate the molecular target of SPN. ResultsIn vivo, we found SPN treatment improved survival rate, alleviated pathological changes through reducing BLM-induced extracellular matrix (ECM) deposition, as well as BLM-induced epithelial-mesenchymal transition (EMT). In vitro, SPN inhibited EMT and lung fibroblast transdifferentiation. Mechanistically, SPN activated the AMPK protein to decrease the abnormally high level of PD-L1. Furthermore, the compound C, known as an AMPK inhibitor, exhibited a significant hindrance to the inhibition of SPN on TGF-β1-caused fibroblast transdifferentiation and proliferation. This outcome could be attributed to the fact that compound C could eliminate the inhibitory effects of SPN on PD-L1 expression. Interestingly, DARTS assay, biofilm interference experiment and molecular docking results all indicated that SPN could bind to AMPK, which suggested that SPN might be a potential agonist targeting AMPK protein. ConclusionAltogether, the results in our work illustrated that SPN promoted AMPK-dependent reduction of PD-L1 protein, contributing to the inhibition of fibrosis progression. Thus, SPN may represent a potential AMPK agonist for PF treatment.

Anti-Inflammatory Mechanism of Ligustrum Lucidum Ait Based on Network Pharmacology and Molecular Docking.

To investigate possible targets of Ligustrum lucidum and its anti-inflammatory mechanism. Core targets were screened by the established networks of component target and potential protein interaction, accompanied by GO and KEGG analyses. The findings were confirmed using molecular docking. eight active components including Quercetin, luteolin, kaempferol and corresponding 163 potential targets, including CCL2, IL6, CXCL8, TNF, IL1B, were identified with a threshold of OB ≥ 30% and DL ≥ 0.18%. 1018 anti-inflammatory targets were acquired, with 101 common targets identified by mutual mapping. 172 nodes and 287 edges are readable in the "component target" visualization network graph. The KEGG analysis identified 20 significantly differentiated signaling pathways, including IL-17, toll-like receptor, TNF, HIF-1, lipid and atherosclerosis. Molecular docking has verified the binding sites and energies of the chemical components in Ligustrum lucidum with key anti-inflammatory proteins, providing a theoretical basis for its clinical use and development.

Regulation of bone and fat balance by Fructus Ligustri Lucidi in ovariectomized mice

Context Fructus Ligustri Lucidi (FLL), a commonly used herb of traditional Chinese medicine (TCM), is the fruit of Ligustrum lucidum Ait. (Oleaceae). The ethanol extract of FLL is a potential candidate for preventing and treating postmenopausal osteoporosis (PMOP) by nourishing the liver and kidneys. Objective This study determines whether an ethanol extract of FLL has anti-osteoporotic effects in ovariectomized (OVX) mice and explores the underlying mechanism. Materials and methods The OVX model of eight-week-old C57BL/6J female mice was taken, and ovariectomy was used as PMOP. Mice were divided into five groups: sham-operated group (n = 10), OVX group (n = 10), OVX + E2 group (n = 10; 0.039 mg/kg), OVX + FLL group (n = 10; 2 g/kg) and OVX + FLL group (n = 10; 4 g/kg). Mice were treated by gavage with FLL or CMCNa once daily for 8 weeks. We harvested uteri, femur, and tibias from mice; bone mineral density (BMD) and bone microstructure were obtained by X-ray absorptiometry and micro-CT. Furthermore, the effect of FLL on the balance of osteoblast and adipocyte differentiation was investigated using bone marrow mesenchymal stem cells (BMMSCs). Results The results indicated that FLL did not affect OVX-induced estradiol reduction. Compared with OVX mice, FLL significantly increased BMD (63.54 vs. 61.96), Conn. D (86.46 vs. 57.00), and left tibial strength (13.91 vs. 11.27), decreased Tb. Sp (0.38 vs. 0.44) and body fat content (4.19% vs. 11.24%). FLL decreased osteoclast activity and enhanced RUNX2 expression; inhibited perilipin peroxisome proliferator-activated receptor gamma (PPARγ) expression and adipocyte differentiation from BMMSCs. Conclusions FLL prevented additional bone loss and improved bone microstructure in OVX mice by modulating bone and fat balance, suggesting that FLL might be a therapeutic agent for PMOP.

Preliminary results on the separation of the different parts of Ligustrum lucidum Ait fruit and the main bioactive compounds analysis

Oleanolic acid (OA) and ursolic acid (UA) possess anti-inflammatory, antioxidative, antiprotozoal, antimutagenic and anticancer properties and are the main bioactive compounds in the fruit of Ligustrum lucidum Ait. The aim of this paper was to study the method of separating the whole fruit into the main ingredients containing OA and UA for the later effective extraction with reduced organic solvents and easy separation and purification. In the present study, the sarcocarps and exocarps were separated from the whole fruits (designated exo-sarcocarps, the mass percentages (w/w, dry weight basis), 33.1%), testae (13.5%) and cores (48.7%) by using methods separated. The contents of OA and UA in whole fruits, exo-sarcocarps, testae and cores were analyzed. The OA and UA extraction yields were highest in exo-sarcocarps vs. yields from whole fruits, testae or cores. The spective yields of OA and UA from exo-sarcocarps were 24.34 ± 2.09 and 7.82 ± 0.09 mg/g; these yields were about 4 times higher than OA yields and about 4 times higher than UA yields from fruit.

Structural characterization and protective effect against renal fibrosis of polysaccharide from Ligustrum lucidum Ait.

Ethnopharmacological relevanceFructus Ligustri Lucidi (FLL), the fruit of Ligustrum lucidum Ait., is a traditional Chinese medicine that has been used for tonifying the kidney and liver for decades. Aim of the studyThis study aimed to explore and identify polysaccharides from FLL and elucidate its protective effect against renal fibrosis. Materials and methodsPolysaccharides were extracted and isolated from FLL. The purified fraction was identified by serial phytochemical work, such as gel-permeation chromatography, ion chromatography, gas chromatography-mass spectrometry, and nuclear magnetic resonance. Mice with unilateral ureteral obstruction (UUO) were applied as a renal fibrosis model. The male UUO mice were pretreated with heteropolysaccharide (Poly) 1 week prior to surgery and continuously treated for 7 days after the operation. Renal fibrosis was assessed by Periodic Acid-Schiff (PAS) staining and Masson's trichrome staining in paraffin-embedded slides. The murine mesangial cells SV40-MES13 upon angiotensin II (Ang II) treatment were developed as an in vitro fibrotic model. The cells were treated by Poly in the presence of Ang II. Molecular expression was detected by RT-PCR, immunoblotting, and immunofluorescence staining. ResultsWe identified a heteropolysaccharide composed of arabinose and galactose (molar ratio, 0.73:0.27) with a predicted chemical structure characterized by a backbone composed of 1,5-α-Araf, 1,3,5-α-Araf, 1,6-α-Galp, and 1,3,6-β-Galp and side chains comprised of T-α-Araf, T-α-Arap, and 1,3-α-Araf. Pretreatment of UUO mice with Poly effectively alleviated glomerulosclerosis and tubulointerstitial fibrosis. Moreover, Poly pretreatment down-regulated the expression of extracellular matrix (ECM) protein fibronectin (FN), profibrotic factor VEGF, proinflammatory cytokines MCP-1 and Rantes in the obstructed kidney. Similarly, the incubation of SV40-MES13 cells with Poly significantly inhibited Ang II-induced elevation in accumulation and expression level of FN and attenuated Ang II-evoked up-regulation in protein expression of MCP-1 and Rantes. ConclusionsOur study isolated and identified a naturally occurring heteropolysaccharide in FLL and revealed its potential in protecting the kidneys from fibrosis.

The Polysaccharide of Ligustrum Lucidum Ait. Protected from Renal Fibrosis by Interfering with Angiotensin II Signaling

BackgroundA number of preclinical advances have indicated the cellular and molecular mechanisms underlying renal fibrosis, such as the signaling of angiotensin II (Ang II), which is a bioactive peptide in renin‐angiotensin system. As one of kidney‐tonifying herbs, Fructus Ligustri Lucidi (FLL), the fruit of Ligustrum lucidum Ait. (Oleaceae) recorded as a tonic and dietary supplement, is a commonly prescribed component in traditional kidney‐tonifying herbal formula. This study aimed to clarify the polysaccharide component in FLL and investigate its potential role in preservation against renal fibrosis.MethodsThe crude polysaccharide of FLL was prepared and deproteinized by Sevag reagent. After lyophilization, the DEAE‐cellulose DE52 column and the Sephacryl S‐200 HR column were used for further isolation and purification of homogenous polysaccharide. The mice with unilateral ureteral obstruction (UUO) underwent daily intraperitoneal injection with FLL polysaccharide starting 7 days before UUO surgery until day 7 after surgery. The murine mesangial SV40‐MES13 cells were treated with vehicle, Ang II (10‐7 M) either with the absence and presence of angiotensin type I receptor blocker (ARB) (10‐6M), or with low dose (50 μg/mL, Poly‐L) and high dose (200 μg/mL, Poly‐H) of FLL polysaccharide.ResultsThe purified homogenous polysaccharide (Poly) was explored from FLL and the carbohydrate content was 97.45%. A hypothetical structure for Poly was proposed considering the comprehensive data of methylation, monosaccharides, and NMR analyses. Treatment of UUO mice with Poly decreased kidney index and ameliorated glomerular sclerosis and interstitial fibrosis, as well as down‐regulated the expression of profibrogenic factor VEGF and matrix fibrotic protein fibronectin (FN). Furthermore, as the same effect to ARB, the pre‐incubation with Poly alleviated the Ang II‐induced up‐regulation of FN in mesangial cells by immunofluoresence staining. The protein expressions of FN and proinflammatory factors like MCP‐1 and Rantes were significantly lower in Poly‐treated cell group than those in Ang II treatment group.ConclusionThe polysaccharide of Ligustrum lucidum Ait. was accurately explored with structure clarification. This polysaccharide component repressed angiotensin II‐induced fibrotic phenotype of renal mesangial cells and exerted beneficial effects on kidney of UUO mice.

Protective Effects of Water Extract of Fructus Ligustri Lucidi against Oxidative Stress-Related Osteoporosis In Vivo and In Vitro.

Fructus Ligustri Lucidi (FLL) is the fruit of Ligustrum lucidum Ait and is a component of many kidney-tonifying traditional Chinese medicine formulae for treating osteoporosis. Accumulating evidence has linked oxidative stress with the progression of bone diseases. The present study aimed to identify the effects of FLL on oxidative stress-related osteoporosis in vivo and in vitro. To construct animal models, we utilized d-galactose (D-gal) injection to induce oxidative stress combined with a low calcium (the exact percentage in the diet was 0.1%) diet. Thirteen-week-old Kunming female mice were gavaged with water extract of FLL for 20 days. Then, eight-month-old Kunming female mice were treated with FLL under standard administration and diet as the aged group. In vitro, MC3T3-E1 cells stimulated by H2O2 were treated with FLL for 24 h. The micro-CT results showed that the modeling approach combining oxidative stress with a low calcium diet caused low conversion type osteoporosis in mice. FLL exerted a prominent effect on preventing osteoporosis by inhibiting oxidative stress, increasing bone mineral density (BMD), improving bone microstructure, and promoting osteoblast proliferation and osteoprotegerin (OPG) protein expression; however, FLL had no therapeutic effect on bone loss in aged mice. In conclusion, FLL showed outstanding anti-bone loss ability both in vivo and in vitro and could probably be developed as a prophylactic agent for osteoporosis.

Effects of air pollution on physiological traits of Ligustrum lucidum Ait. leaves in Luoyang, China

Leaves of trees experience the maximum brunt of exposure and undergo certain changes in physiological traits responding to air pollution, and then, the specific leaf traits can be the indicators of air pollution in an area. However, due to the diversity of sources, the composition of air pollutants is very complex. This makes it difficult to predict air pollution using physiological differentiation of leaves. The purpose of this investigation was to examine potential of Ligustrum lucidum Ait. leaf measurement as a method to predict the air pollutants in Luoyang, China. Leaves of roadside L. lucidum were studied from the city center with serious air pollution to relatively unpolluted areas. Leaf size, stomatal traits, and non-structural carbohydrate were measured. The particulate and gaseous pollutants (including sulfur dioxide, nitrogen dioxide, carbon monoxide, and ozone) were investigated too. The results showed that the leaf area and soluble sugar content decreased, while the aspect ratio of leaves increased in heavily polluted areas. As pollution increased, the stomatal traits in different crown positions were changed differently. No significant correlation was found between ozone content and the measured traits of leaves. The responses found in the physiological differentiation of the leaves reflect acclimation to air pollution. The soluble sugar content of the leaves could be used to indicate the short-term stress of air pollution, the area, and aspect ratio of leaves are indicative of the long-term stress due to air pollution. Therefore, physiological traits of L. lucidum leaves appeared to be significant predictive factors for the air pollutants in urban areas.

Ligustroflavone ameliorates CCl4-induced liver fibrosis through down-regulating the TGF-β/Smad signaling pathway.

Liver fibrosis is a pathological process characterized by excess deposition of extracellular matrix (ECM) that are mainly derived from activated hepatic stellate cells. Previous studies suggested that ligustroflavone (LF) was an ingredient of Ligustrum lucidum Ait. with activities of anti-inflammation and anti-oxidation. In this study, we investigated whether LF had any effect on liver fibrosis. In our study, we established a mouse model of carbon tetrachloride (CCl4)-induced liver fibrosis and used TGF-β1-stimulated human hepatic stellate cell line (LX-2) to explore the effect of LF and associated underlying mechanism. LF was used in vivo with low dose (L-LF, 5 mg·kg-1, i.p., 3 times each week) and high dose (H-LF, 20 mg·kg-1, i.p., 3 times each week) and in vitro (25 μmol·L-1). Histopathological and biochemical assays investigations showed that LF delayed the formation of liver fibrosis; decreased AST, ALT activities and increased Alb activity in serum; decreased MDA level, Hyp content and increased GSH-Px concentration, SOD activity in liver tissues. Moreover, immunohistochemical, immunofluorescent and Western blot results showed that LF reduced the expressions of hepatic stellate cells specific marker proteins, including collagen I and α-SMA in vivo and in vitro. In addition, LF markedly suppressed TGF-β1-upregulated protein expressions of TβR I, TβR II, P-Smad2, P-Smad3 and Smad4 in LX-2 cells. Taken together, these findings demonstrated LF could decrease histopathological lesions, ameliorate oxidative injury, attenuate CCl4-induced liver fibrosis, which may be associated with down-regulating the TGF-β/Smad signaling pathway.

A Pentacyclic Triterpene from Ligustrum lucidum Targets γ-Secretase

Amyloid-beta peptides generated by β-secretase- and γ-secretase-mediated successive cleavage of amyloid precursor protein are believed to play a causative role in Alzheimer's disease. Thus, reducing amyloid-beta generation by modulating γ-secretase remains a promising approach for Alzheimer's disease therapeutic development. Here, we screened fruit extracts of Ligustrum lucidum Ait. (Oleaceae) and identified active fractions that increase the C-terminal fragment of amyloid precursor protein and reduce amyloid-beta production in a neuronal cell line. These fractions contain a mixture of two isomeric pentacyclic triterpene natural products, 3-O-cis- or 3-O-trans-p-coumaroyl maslinic acid (OCMA), in different ratios. We further demonstrated that trans-OCMA specifically inhibits γ-secretase and decreases amyloid-beta levels without influencing cleavage of Notch. By using photoactivatable probes targeting the subsites residing in the γ-secretase active site, we demonstrated that trans-OCMA selectively affects the S1 subsite of the active site in this protease. Treatment of Alzheimer's disease transgenic model mice with trans-OCMA or an analogous carbamate derivative of a related pentacyclic triterpene natural product, oleanolic acid, rescued the impairment of synaptic plasticity. This work indicates that the naturally occurring compound trans-OCMA and its analogues could become a promising class of small molecules for Alzheimer's disease treatment.

Biomagnetic monitoring combined with support vector machine: a new opportunity for predicting particle-bound-heavy metals

Biomagnetic monitoring includes fast and simple methods to estimate airborne heavy metals. Leaves of Osmanthus fragrans Lour and Ligustrum lucidum Ait were collected simultaneously with PM10 from a mega-city of China during one year. Magnetic properties of leaves and metal concentrations in PM10 were analyzed. Metal concentrations were estimated using leaf magnetic properties and meteorological factors as input variables in support vector machine (SVM) models. The mean concentrations of many metals were highest in winter and lowest in summer. Hazard index for potentially toxic metals was 5.77, a level considered unsafe. The combined carcinogenic risk was higher than precautionary value (10−4). Ferrimagnetic minerals were dominant magnetic minerals in leaves. Principal component analysis indicated iron & steel industry and soil dust were the common sources for many metals and magnetic minerals on leaves. However, the poor simulation results obtained with multiple linear regression confirmed strong nonlinear relationships between metal concentrations and leaf magnetic properties. SVM models including leaf magnetic variables as inputs yielded better simulation results for all elements. Simulations were promising for Ti, Cd and Zn, whereas relatively poor for Ni. Our study demonstrates the feasibility of prediction of airborne heavy metals based on biomagnetic monitoring of tree leaves.

Chemical composition and glucose uptake effect on 3T3-L1 adipocytes of Ligustrum lucidum Ait. flowers

Ligustri lucidi Fructus is a traditional Chinese medicine and possesses various bioactivities, including hypoglycemic effect. Ligustrum lucidum Ait flowers are poorly investigated. Thus, we hypothesized that L. lucidum flowers also could have hypoglycemic effect. Chemical composition and glucose uptake effect of L. lucidum flowers on 3T3-L1 adipocytes were investigated. In this study, the components of L. lucidum flowers were investigated by various chromatographic and spectroscopic methods and the effects of L. lucidum flowers on the induction of glucose uptake were investigated by 3T3-L1 adipocytes. Seven compounds were isolated and identified from L. lucidum flowers, including ursolic acid (1), kaempferol-7-O-α-l-rhamnoside (2, KR), β-sitosterol (3), β-daucosterol (4), kaempferitrin (5, KF), 10-hydroxy oleuropein (6, HO), and kaempferol-3-O-β-d-glucopyranoside-7-α-l-rhamnopyranoside (7, KGR). The results of glucose uptake showed that total extract (TE), KR, KF and HO from L. lucidum flowers significantly enhanced glucose uptake. This study demonstrated that compounds 2, 5-7 were isolated for the first time from this plant and compounds 2 and 7 were isolated for the first time in genus Ligustrum, L. lucidum flowers, KR, KF and HO may possess potential hypoglycemic effect.

Ethanol extract of Ligustrum lucidum Ait. leaves suppressed hepatocellular carcinoma in vitro and in vivo

BackgroundThe present study investigated the pharmacological activity and mechanism of ethanol extract of Ligustrum lucidum Ait. leaves (EEL) on HCC.MethodsCell viability was determined using cell counting kit-8 (CCK-8) assay. The effects of EEL on cellular biological activities were analyzed by flow cytometry (FCM), cell wound scratch assay and transwell assay. The expression levels of related mRNA and protein were determined by performing quantitative real-time PCR (qRT-PCR), Western blotting assay and immunocytochemistry. Methylation-specific PCR (MSP) was carried out to investigate the possible mechanism underlying the DNA methylation of PTEN.ResultsEEL showed cytotoxicity to both Bel-7402 and Huh-7 cell lines. We also found that EEL enhanced the apoptosis of Bel-7402 and Huh-7 cells by regulating the expressions of Bcl-2 associated X (Bax), B cell lymphoma 2 (Bcl-2) and Cytochrome-C and the activity of caspase-3 and therefore promoted cell cycle arrest. Moreover, EEL also suppressed cell migration and invasion. EEL increased the expression of tissue inhibitor of metalloproteinases 2 (TIMP2) but decreased the expressions of matrix metalloproteinase2 (MMP2) and MMP9. Furthermore, EEL inhibited the phosphorylation of PI3K/Akt pathway. MSP results showed that EEL promoted the demethylation of PTEN, suggesting that the inactivation of PI3K/Akt may be related to DNA de-methylation of PTEN. In addition, EEL inhibited the tumor growth of HCC in vivo.ConclusionsEEL exerted anti-tumor effect on HCC in vitro and in vivo. EEL mediated by the inhibition of PI3K/Akt may be closely related to DNA de-methylation of PTEN. Thus, EEL could be used as a potential anti-cancer therapeutic agent of HCC.

Four spices prevent mice from contracting Salmonella enterica serovar Typhimurium.

Ligustrum lucidum Ait (LL), Lysimachia christinae Hance (LC), Mentha piperita Linn (MP), and Cinnamomum cassia Presl (CC) are common spices used in Asia. The present study investigated the anti-Salmonella effects of the four spices using aqueous extracts. The amount of phenolic acids and flavonoids in each spice aqueous extract was determined as indicators of purity. Mice were pretreated with LL, LC, MP or CC aqueous extract for 7 days. Following infection with Salmonella enterica serovar Typhimurium strain ST21 (ST21), the aqueous extract of each spice was subsequently administered for 4 days. ST21 infected mice had lower body weight compared with the control group. The administration of spice aqueous extracts significantly increased body weight following infection. ST21 infection increased the fecal ST21 counts compared with the control group; however, following spice aqueous extract treatments, ST21 counts significantly decreased. The spice treatments also significantly reduced ST21 count in blood and the organs. Notably, ST21 infection increased interferon (IFN)-γ and interleukin (IL)-6 levels in serum whilst spice treatments reduced these cytokines. In the spleen, spice treatment significantly lowered IFN-γ, IL-6, IL-1β, and tumor necrosis factor (TNF)-α levels, but increased IL-12 levels. ST21 infection stimulated the production of immunoglobulin (Ig)A and IgM in serum whilst spice aqueous extract treatment significantly decreased these levels. In summary, LL and MP aqueous extract treatments had the most significant effect in protecting against ST21 infection. Results of the RAW 264.7 cell infection model suggested that the mechanisms involved in the anti-ST21 effect of each spice were individually different. All four aqueous extracts demonstrated different mechanisms in attenuating ST21 invasion with the protective effect of LC aqueous extract potentially involving TNF-α expression. The present findings suggested that the four spices may be considered as potent functional foods due to their anti-Salmonella effects.

Estimation of urban POP and emerging SVOC levels employing Ligustrum lucidum leaves

Many persistent organic pollutants (POPs) have been banned in many countries including Argentina after enforcing the Stockholm Convention in 2014, while other emerging semi-volatile organic contaminants (SVOCs) are considered to enter the list due to their known environmental persistence and toxicity. However, there is still very little information regarding the distribution of these chemicals in the environment in developing countries. To address this issue, we employed leaves of Ligustrum lucidum Ait. as a passive monitor to estimate urban levels of polychlorinated biphenyls, brominated flame retardants and hexachlorobenzene (PCBs, BFRs, and HCB, respectively) considering three different land use areas in Córdoba city (Argentina). We found higher PCB values in urban and industrial areas, which could be attributed to local emission sources as well as a long-range transport of lightweight compound. BFRs were more abundant in the urban areas indicating that their main emission source is the volatilization from polymeric materials. HCB, on the other hand, was equally distributed at the three sampling areas. Overall, POP and SVOC levels were similar or even lower than some other urban environments and even comparable with remote places elsewhere.

Compatibility effects of ginseng and Ligustrum lucidum Ait herb pair on hematopoietic recovery in mice with cyclophosphamide-induced myelosuppression and its material basis

BackgroundGinseng (G) and Ligustrum lucidum Ait (LLA) are core traditional Chinese medicines in treating myelosuppression formula. The present study was designed to profile effect of G and LLA herb pair (G-LLA) on myelosuppressed mice. MethodsThe mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (Cy). Hematopoietic function of bone marrow was measured by hemopoietic progenitor cell culture and peripheral blood count, and serum hemopoietic factors were tested by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. HPLC was used to measure 20 potential chemical components related to myelosuppression, including ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd, Rk3, Rh4, 20 (S)-Rg3, 20 (R)-Rg3, Rk1, Rg5, salidroside, and so on. ResultsG, LLA, and G-LLA improved the amount of peripheral blood cells and bone marrow cells of myelosuppressed mice (P < 0.01). They significantly increased the colony quantity of colony-forming unit–granulocyte macrophage, burst-forming unit−erythroid, colony-forming unit−erythroid, and colony-forming unit−megakaryocyte and amount of G2/M and S phase cells (P < 0.01). They also significantly decreased the amount of hematopoiesis-related cytokines (P < 0.01). The content of chemical components in G-LLA changed, and the change of rare saponin was the most obvious. ConclusionThese results show that G-LLA herb pair might produce synergistic or complementary compatibility effects on bone marrow suppression after chemotherapy. It suggests that the substance basis of G-LLA for treating bone marrow suppression may be effective chemical components.

EFFECTS OF SHORT-TERM NITROGEN AND PHOSPHORUS ADDITION ON ROOT BIOMASS AND MORPHOLOGICAL CHARACTERISTICS OF LIGUSTRUM LUCIDUM AIT IN NEW RECLAMATION COASTAL FIELD

EFFECTS OF SHORT-TERM NITROGEN AND PHOSPHORUS ADDITION ON ROOT BIOMASS AND MORPHOLOGICAL CHARACTERISTICS OF LIGUSTRUM LUCIDUM AIT IN NEW RECLAMATION COASTAL FIELD