A New Sesquiterpene Polyol Ester from Celastrus Angulatus

A new sesquiterpene polyol ester, angulatin D, has been isolated from the root bark of Celastrus angulatus. The chemical structure is elucidated on the basis of spectral analysis.

A New Sesquiterpene from Celastrus Angulatus

One new sesquiterpene polyol eater named angulatin C was isolated from the root bark of Celastrus angulatus along with a known compound, angulatin B. Their structures were elucidated on the basis of spectral analysis.

Celastrus paniculatus oil ameliorates synaptic plasticity in a rat model of attention deficit hyperactivity disorder

Objective:To explore the effect and mechanism of action of Celastrus paniculatus oil on the treatment of perinatal rats with attention deficit hyperactivity disorder.Methods:In the perinatal stage,the rats were either isolated or administered with lead acetate to establish an animal model of attention deficit hyperactivity disorder.Atomoxetine served as the reference standard.Animals’behaviours were assessed through Y-maze,novel object preference,fear conditioning and residentintruder aggression tests.Oxidative stress parameters,bioamine concentration(dopamine,noradrenaline and 5-hydroxytryptamine),nerve growth factor,interleukin-6,nuclear factor-κB,and tumour necrosis factor(TNF)-αwere estimated.Synaptophysin immunohistochemical assay was performed.Results:Celastrus paniculatus oil significantly improved behavioural parameters in Y maze,novel object preference,discrimination index,fear conditioning and resident intruder aggressive tests.The treatment groups showed a decrease in malondialdehyde level.Changes in the levels of dopamine,noradrenaline,and serotonin were restored by Celastrus paniculatus oil.Celastrus paniculatus oil increased nerve growth factor and decreased interleukin-6,nuclear factor-κB,and TNF-α.Synaptophysin immunoreactivity was also improved by Celastrus paniculatus oil with alleviated reactive gliosis,degeneration,and vascular proliferation.Conclusions:This research shows the therapeutic potential of Celastrus paniculatus oil for the treatment of attention deficit hyperactivity disorder.

A Novel Sesquiterpene Polyol Ester from Celastrus Angulatus and its Antitumor Activities

A new sesquiterpene polyol ester was isolated from the leaves of Celastrus angulatus. Its structure was determined as 1 (beta), 2 (beta), 9 (a) -triacetoxy-8 (a) -((a) -hydroxyl-isobutyryloxy)-15-benzoyloxy-4 (a) -hydroxy-dihydroagarofuran by means of NMR, MS and IR spectral analysis. Preliminary biological study on antitumor activity revealed that this compound showed strong nonselective cytotoxicity against four of the NCI panel cell lines.

The petroleum ether fraction of Celastrus paniculatus Willd. seeds demonstrates antiarthritic effect in adjuvant-induced arthritis in rats

Objective:Celastrus paniculatus(CP)Willd.is a plant indigenous to India with medicinal properties.It is used in the ayurvedic treatment of inflammatory ailments such as rheumatoid arthritis.The present study was designed to investigate the therapeutic efficacy of the petroleum ether fraction(PCP)obtained from CP seeds on adjuvant-induced arthritis in rats.Methods:Arthritis was induced in SpragueeDawley rats by immunization with Freund’s complete adjuvant(FCA).Arthritis severity was evaluated by arthritis score,paw volume,tibiotarsal joint thickness,body weight,dorsal flexion pain,motility test,stair climbing ability and index of thymus and spleen.Moreover,histopathology of knee joints supported by haematological analysis was used to assess the anti-arthritic action of PCP.The levels of superoxide dismutase(SOD),glutathione(GSH),catalase(CAT),nitric oxide(NO)and malondialdehyde(MDA)in liver were also assessed.Serum samples were collected for estimation of aspartate transaminase(AST),alanine transaminase(ALT)and alkaline phosphatase(ALP).In addition,cytokines such as tumour necrosis factor-alpha(TNF-a)and interleukin-6(IL-6)were estimated in plasma.Results:PCP significantly alleviated arthritic progression with regards to paw swelling,arthritic score,immune organ indices,hyperalgesic effect and body weight.This phenomenon was correlated with significant suppression of overproduction of inflammatory cytokines(TNFa and IL-6),oxidant stress markers(MDA and NO)and cellular enzyme(AST,ALT and ALP)levels versus arthritic rats without treatment.Moreover,PCP restored the decreased levels of SOD,CAT and GSH.Conclusion:Our results suggest that the anti-arthritic properties of PCP may be due to immunosuppressive effects,cytokine regulation,antioxidant effects and bone-protective activities.

Two New Sesquiterpenes from Celastrus Paniculatus.Subsp.Paniculatus

Two new beta-dihydroagarofuran sesquiterpene polyol esters were isolated from Celastrus paniculatus.subsp.paniculatus. Their Structures were deduced on the basis of spectral analyses including 2D NMR. CO2 supercritical fluid was used in separation.

A NEW SESQUITERPENE FROM CELASTRUS ROSTHONIANUS

A new sesquitcrpene was isolated from Celastrus rothornianus, its structure was clucidated mainly on the basis of spectral analysis.

A NEW ALKALOID FROM CELASTRUS ANGULATUS

A new β-dihydroagarofuran alkaloid Ⅰ was isolated from Celastrus angulatus.The structure was established on the basis of 1D and 2D NMR analysis,and x-ray diffraction.

Study on effect and mechanism of Celastrus Orbiculatus total terpenoids in regulating lipid metabolism

Objective To investigate the effect and mechanism of Celastrus Orbiculatus total terpenoids on lipid metabolism in hyperlipidemia mice.Methods ICR mice were selected as investigated subject.The hyperlipemia mice models were made with feeding high-fat forage and were randomly divided into six groups:the normal group,the model group,the positive control group(treated with simvastatin)and the three groups treated with Celastrus Orbiculatus total terpenoids with low,medium and high dosage,respectively.Each group included eight mice.The control group was fed normal forage,but other groups were fed high fat forage.All groups were allowed to drink water freely.Since the first day when the models were made,intragastric administration had been adopted.The normal group was fed normal forage without intragastric administration;the model group was received physiological saline 20 mL·kg-1·d-1 with intragastric administration;the positive control group received simvastatin 2.6 mg·kg-1·d-1 with intragastric administration;the three treated groups received Celastrus Orbiculatus total terpenoids 60 mg·kg-1·d-1,120 mg·kg-1·d-1,200 mg·kg-1·d-1 with intragastric administration respectively.Each group was weighed once a week.On the basis of establishing hyperlipidemia mice model,blood lipids,lipid metabolic enzyme,antioxidative capacity were investigated after 21 days feeding of high-fat forage.Results Compared with model group,TC and TG in mice treated with Celastrus Orbiculatus total terpenoids all reduced and HDL-C raised obviously(P<0.01).Celastrus orbiculatus total terpenoids was shown to decreased MDA content in both serum and liver,increased serum SOD activity and inhibited the activity of the cholesteryl ester transfer protein(CETP).Conclusions Celastrus Orbiculatus total terpenoids could remarkably modulate the lipid metabolic disorder in hyperlipidemia mice,and has a certain regulating function on lipoprotein,inferring that it could reduce the occur of atherosclerosis.The mechanism of regulating lipid metabolism might be related with decreasing the activity of CETP and increasing antioxidative capacity.

INVESTIGATION OF CHEMICAL CONSTITUENT FROM CELASTRUS ANGULATUS

Three new β-dihydroagarofuran sesquiterpene polyol esters,one alkaloid and two non-alkaloids,were isolated from the MeOH extract of seed oil of Celastrus angulatus. Their structures were elucidated on the basis of UV,IR,mass,NOE difference,~1H-NMR,^(13)C- NMR and ~1H-^(13)C long-range correlation spectroscopy.

AN IHSECTICIDAL SESQUITERPENE FROM CELASTRUS ANGULATUS

An insecticidal sesquiterpene polyol ester,angulatin A,was isolated from the root bark of Celastrus angulatus.Its structure was established as 1α,2α-diacetoxy-8β,15-diisobutyryloxy-9α- benzoyloxy-4β,6β-dihydroxydihyclro-β-agarofuran by spectroscopic methods.

Erratum to "Celastrus paniculatus oil ameliorates synaptic plasticity in a rat model of attention deficit hyperactivity disorder"

In the article“Celastrus paniculatus oil ameliorates synaptic plasticity in a rat model of attention deficit hyperactivity disorder”published on pages 105-114,Issue 3,Volume 11 of Asian Pacific Journal of Tropical Biomedicine,Figure 5 was incorrectly published.

Apoptosis of Hela cell induced by Celastrus orbiculatus Thunb extract and primary mechanisms

Objective： The apoptosis of Hela cells induced by ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb was studied in order to assess its antitumor effect. Methods： Hela cells were cultured in vitro and treated by a series of concentrations of ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb. Cell proliferation was detected based on MTT assay. Quantity of apoptosis were observed and analyzed by flow cytometry with Annexin V and propidium iodide double staining. P53 gene expression was detected by flow cytometry. Results： The proliferation of Hela cells was obviously inhibited by 15, 30, 60 and 120 IJg/mL extract of Celastrus orbiculatus Thunb and apoptosis of Hela was induced by dosed dependent manner. P53 gene showed increasing tendency when treated by 60-480 IJg/mL extract of Celas- trus orbiculatus Thunb. Conclusion： The ethyl acetate extract and n-butanol extract of Celastrus orbiculatus Thunb could induce apoptosis of Hela gastric cancer cells by dose dependent manner, which maybe one of the important mechanisms of Celastrus orbiculatus Thunb＇s anticancer effects. P53 protein expression in Hela was up-regulated by Celastrus orbiculatus Thunb, which maybe one of the molecular mechanisms involved in the anticancer and proapoptotic effect of Celastrus or- biculatus Thunb.

Study on Extraction Techniques of Celastrol from Celastrus orbiculatus Thunb.

[Objectives]This study was conducted to investigate the extraction technology of celastrol from Celastrus orbiculatus Thunb.[Methods]Solvent ultrasonic extraction was selected,and with the content of celastrol as the evaluation index,the effects of different solvents,extraction time,temperatures and material-to-liquid ratios on the extraction rate of celastrol were investigated by single factor and orthogonal experiments.[Results]The optimal extraction conditions were as follows:a solvent ratio of petroleum ether to ethyl acetate at 1∶1,a ratio of solvent to material at 10∶1(v/w),extraction time of 30 min,and an extraction temperature at 30℃.[Conclusions]This method has high extraction rate,and is simple and feasible.

Celastrus orbiculatus Extract Inhibits Immune Inflammatory Thrombotic State of B-Lymphoma

Objective:To investigate the inhibitory effect of Celastrus orbiculatus extracts(COE)on the proliferation of lymphoma cells and the immune regulation ability on inflammation and thrombophilia in vivo.Methods:The 38B9 lymphoma cells were treated with COE(160μg/mL)and CTX(25μmol/L).The apoptosis rate and cell cycle of each group were detected by flow cytometry.The secretion of inflammatory factors,including interleukin(IL)-6,IL-10,and tumor necrosis factorα(TNF-α),in cell supernatant was detected by enzyme-linked immunosorbent assay(ELISA).In vivo,BALB/c mice were subcutaneously injected with 38B9 lymphoma cells to establish lymphoma model.COE(3 mg·kg^(-1)·d^(-1))and CTX(40 mg·kg^(-1)·d^(-1))were administered to the model mice,respectively.The expression of plasma inflammatory factors(IL-6,IL-10 and TNF-α)and thrombus indexes,including D-dimer(D-D),von Willebrand factor(vWF)and tissue factor(TF),were detected by ELISA before tumor bearing(1 d),after tumor formation(14 d)and after intervention(21 d).PicoGreen dsDNA was used to detect the level of serum neutrophil extracellular traps(NETs).Flow cytometry was used to detect the expression of platelet activation marker calcium-dependent lectin-like receptor 2(CLEC-2).The tumor growth and survival of mice were recorded.Results:The 38B9 lymphoma cells were apoptotic after the intervention of COE and CTX.The ratio of Gz-M phase cells decreased in COE intervented cells compared with the control cells(P<0.05),and S phase cells decreased in CTX intervented cells(P<0.05).Also,the secretion level of IL-6 was significantly reduced after COE or CTX intervention(P<0.05),and IL-10 was significantly increased(P<0.05).Furthermore,the tumor mass was reduced,and the median survival time was longer in COE and CTX intervented tumor-bearing mice than in non-intervented mice.The significantly lower levels of TNF-α,IL-6,NETs,TF,DD and CLEC-2,as well as higher IL-10 were observed in COE and CTX treatment mice in comparision with the control mice(P<0.05).Conclusion:COE has a mild and stable anti-tumor effect,which can reduce the secretion of inflammatory factors by lymphoma cells and regulate thrombophilic state caused by tumor inflammatory microenvironment.

Celastrus orbiculatus extract induces mitochondrial-mediated apoptosis in human hepatocellular carcinoma cells

OBJECTIVE： To investigate the apoptotic effects and underlying molecular mechanisms of Celastrus orbiculatus （C. orbiculatus） extract in human hepa- tocellular carcinoma cells. METHODS： Human hepatocellular carcinoma cells （HCCLM6） were treated with C. orbiculatus extract （COE） at different nontoxic concentrations （10, 20, 40, 80, and 160 IJg/mL）. The effect of COE on HC-CLM6 viability was examined using 3-（4,5-dimethyl- thiazol-2-yl）-2,5-diphenyl tetrazolium bromide （MTT） assays. Cellular apoptosis following COE treatment was assessed by flow cytometry and western blot analysis. RESULTS： COE significantly inhibited cell viability and induced apoptosis of HCCLM6 cells in a dose-dependent manner. Apoptosis was accompa- nied by increased Bax expression and decreased Bcl-2 expression. In addition, COE treatment led to the release of cytochrome c, activation of cas- pase-3, and cleavage of poly （ADP-ribose） poly- merase （PARP）. Furthermore, activation of extracel- lular signal-regulated kinase （ERK）, p38 kinase, and c-Jun N-terminal kinase （JNK） phosphorylation, and down-regulation of Akt phosphorylation was ob- served. CONCLUSION： COE induces mitochondrial-mediat- ed, caspase-dependent apoptosis in HCCLM6 cells, which might be attributed to the activation of mito- gen-activated protein kinase （MAPK） and inhibition of Akt signaling pathways. These data suggest that COE may be a potential treatment for human hepa- tocellular carcinoma.

Celastrus Orbiculatus Extract Inhibits Tumor Angiogenesis by Targeting Vascular Endothelial Growth Factor Signaling Pathway and Shows Potent Antitumor Activity in Hepatocarcinomas in Vitro and in Vivo

Objective： Ce/astrus orbicu/atus Thunb. has been used for thousands of years in China as a remedy against cancer and inflammatory diseases. This study aims to investigate whether C. orbiculatus extract （COE） could inhibit angiogenesis, which is the pivotal step in tumor growth, invasiveness, and metastasis. Methods： In this study, the extract from the stem of C. orbiculatus was used. Mouse hepatic carcinoma cells （Hepat-6） were treated with COE in different nontoxic concentrations （10, 20, 40, 80, and 160 μg/mL）. The mRNA and protein expression levels of vascular endothelial growth factor （VEGF） were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot, respectively; the active fractions were further tested on C57BL/6 mice and human umbilical vein endothelial cells （HUVEC） for any antiangiogenic effects. Results： COE significantly inhibited proliferation and induced apoptosis in Hepal-6 cells and inhibited VEGF expression at both mRNA and protein levels. Furthermore, this agent inhibited the formation of the capillary-like structure in primary cultured HUVEC in a dose-dependent manner. In vivo, COE significantly reduced the volume and weight of solid tumors with low adverse effects and decreased tumor angiogenesis. Conclusions： In summary, COE could be used to treat hepatic carcinoma. The mechanisms of the antitumor activity of COE may be due to its effects against tumor angiogenesis by targeting the VEGF protein.

Extracts of Celastrus Orbiculatus Inhibit Cancer Metastasis by Down-regulating Epithelial-Mesenchymal Transition in Hypoxia-Induced Human Hepatocellular Carcinoma Cells

Objective: To evaluate the effects of Celastrus Orbiculatus extracts(COE) on metastasis in hypoxiainduced hepatocellular carcinoma cells(Hep G2) and to explore the underlying molecular mechanisms. Methods:The effect of COE(160, 200 and 240 μg/mL) on cell viability, scratch-wound, invasion and migration were studied by 3-4,5-dimethyl-2-thiazolyl-2,5-diphenyl-2-H-tetrazolium bromide(MTT), scratch-wound and transwell assays, respectively. Co Cl2 was used to establish a hypoxia model in vitro. Effects of COE on the expressions of E-cadherin, vimentin and N-cadherin were investigated with Western blot and immuno?uorescence analysis,respectively. Results: COE inhibited proliferation and metastasis of hypoxia-induced hepatocellular carcinoma cells in a dose-dependent manner(P<0.01). Furthermore, the expression of epithelial-mesenchymal transition(EMT) related markers were also remarkably suppressed in a dose-dependent manner(P<0.01). In addition, the upstream signaling pathways, including the hypoxia-inducible factor 1α(Hif-1α) and Twist1 were suppressed by COE. Additionally, the Hif-1α inhibitor 3-5'-hydroxymethyl-2'-furyl)-1-benzylindazole(YC-1), potently suppressed cell invasion and migration as well as expression of EMT in hypoxia-induced Hep G2 cells. Similarly, the combined treatment with COE and YC-1 showed a synergistic effect(P<0.01) compared with the treatment with COE or YC-1 alone in hypoxia-induced Hep G2 cells. Conclusions: COE signi?cantly inhibited the tumor metastasis and EMT by suppressing Hif-1α/Twist1 signaling pathway in hypoxia-induced Hep G2 cell. Thus, COE might have potential effect to inhibit the progression of Hep G2 in the context of tumor hypoxia.

Inhibition of Celastrus orbiculatus Extracts on VEGF Expression in Hepatoma Cells of Mice

Objective To discuss the antitumor mechanism preliminarily by observing effects of Celastrus orbiculatus extracts(COE) on vascular endothelial growth factor(VEGF) mRNA and protein expression in hepatoma(Hepa1-6) cells of mice.Methods Hepa1-6 cells were treated with COE at different nontoxic concentration(0,10,20,40,80,and 160 μg/mL) for 16 h.The mRNA and protein expressions of VEGF were detected by reverse transcription-PCR and Western Blotting,respectively.Results COE significantly inhibited VEGF expression at both mRNA and protein levels in a dose-dependent manner.Conclusion COE can inhibit VEGF expression in Hepa1-6 cells,therefore suggest that VEGF could be chosen as an therapeutic target for COE in the context of cancer chemoprevention and anticancer therapy.

Celastrus orbiculatus Extracts Inhibit Human Hepatocellular Carcinoma Growth by Targeting mTOR Signaling Pathways

Objective:To characterize the molecular mechanism underlying the antineoplastic activity of Celastrus orbiculatus Thunb.extracts(COE).Methods:The human hepatocellular carcinoma HepG2 cells with mammalian target of rapamycin(mTOR)knockdown expressed(HepG2/mTOR-)were constructed using molecular biological technology.In vitro,the HepG2/mTOR-cells were treated with COE at various concentrations(10,20,40,80,160 and 320μg/mL).Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assays.According to the half-maximal inhibitory concentration(IC50)value(140 mg/L),the concentrations of COE in the subsequent experiment was set to alleviate cytotoxicity.The HepG2/mTOR-cells were divided into 5 groups:negative control(untreated),COE treatment groups(40,80,120 mg/L COE)and positive control group(cisplatin,DDP,2 mg/L),respectively.Wild-type HepG2 cells were used as a blank control.The effects of COE on the cell apoptosis were analyzed by flow cytometry and transmission electronic microscopy(TEM),respectively.The protein expression levels of mTOR signal pathways were determined by Western blotting.In vivo,HepG2/mTOR-cells(2×106 cell/mice)were subcutaneously injected into the right flank of nude mice.Thirty-six female nude mice were randomly assigned to 6 groups according to body weight(6 mice per group)as follows:solvent vehicle control,Banmao Capsule treated group(BM,195 mg/kg),Tegafur,Gimeracil and Oteracil Potassium Capsules(10 mg/kg)treated group,and different dosages of COE(10,20,40 mg/kg)groups.Tumor growth was monitored and immunohistochemical staining was used to examine the expression of apoptosis-related proteins in tumor tissues.Results:COE inhibited the proliferation significantly in a concentration-dependent manner in HepG2/mTOR-cells(P<0.01).COE significantly induced the apoptosis of HepG2/mTOR-cells(P<0.01),and the apoptotic bodies can be observed under TEM.COE significantly inhibits the proteins expression of mTORrelated signal pathways.In vivo,COE significantly inhibited tumor growth in nude mice(P<0.01).Moreover,the results showed that COE down-regulated the expression of Bcl-2 and Bcl-xL,and up-regulated the levels of Bax and caspase-3 protein(P<0.01).Conclusion:COE was a potential chemotherapeutic drug in HCC treatments via targeting mTOR signal pathway.