Tormentic Acid Research Articles

Anti-hepatocellular carcinoma activity of tormentic acid derived from suspension cells of Eriobotrya japonica (Thunb.) Lindl.

Tormentic acid (TA), which is the main pentacyclic triterpene secondary metabolite formed by Eriobotrya japonica (Thunb.) Lindl., was purified by preparative HPLC and characterized by UPLC-MS. The inhibitory activity of the purified material towards hepatocellular carcinoma cells (HepG2, Bel-7405, Sk-hep-1) was evaluated based on proliferation, tumorigenesis, transformation, invasion and migration, as well as its ability to induce apoptosis in these cells. At concentrations of 22.5 μg/mL or more, TA led to considerable decreases in cell viability, colony formation and cell migration. TA treatment also induced apoptosis in the three hepatocellular carcinoma cell lines evaluated in this study. These effects were related to changes in the expression levels of Caspase3 and poly ADP-ribose polymerase, therefore highlighting the potent anti-cancer activity of TA towards all three of these cell lines.

Tormentic Acid Inhibits IL-1β-Induced Inflammatory Response in Human Osteoarthritic Chondrocytes

Tormentic Acid Inhibits IL-1β-Induced Inflammatory Response in Human Osteoarthritic Chondrocytes

Extraction of antioxidant and antiproliferative ingredients from fruits of Rubus chingii Hu by active tracking guidance.

Rubus chingii Hu, namely "Fu-pen-zi" in Chinese, has been used as a functional food in China for a long time. This study aims to identify its bioactive constituents with antioxidant and anti-tumor properties. R. chingii was extracted with 95% ethanol and then partitioned into four fractions: petroleum ether fraction, ethyl acetate fraction, n-butanol fraction, and water fraction. Results showed that the ethyl acetate fraction had the strongest antioxidant activity and cytotoxicity against human cancer cell lines (HepG-2, Bel-7402, A549 and MCF-7). Therefore, four compounds were isolated from the ethyl acetate fraction, and they were identified as ent-16α,17-dihydroxy-kauran-19-oic acid, tormentic acid, oleanolic acid and β-daucosterol, the first two of which were isolated and identified from R. chingii for the first time. In particular, tormentic acid exhibited excellent cytotoxicity activities against human tumor cell lines. The results obtained in this work might contribute to the understanding of biological activities of R. chingii and further investigation on its potential application is valued for food and drugs.

A Review on Secondary Metabolites of Rosa laevigata Michaux: An Important Medicinal Plant

Rosa laevigata is a white aromatic rose inhabitant to Southern China and Taiwan and growing offensively as invasive in the United States of America. It is herbaceous climbing shrub, growing over the other shrubs and reaching upto 5-10 metre in height. In 1780’s, it gained its English name Cherokee rose from America. This review was aimed to analyze various active secondary metabolites of Rosa laevigata with their medicinal value. The study infers that plant possess novel secondary metabolites i.e., polysaccharides, flavonoids, steroids, tannins, laevigatins E, F, G, triterpenoids, 11α-hydroxytormentic acid, 2α-methoxyursolic acid, 6-methoxy-β-glucopyranosyl ester, tormentic acid and 5α-diol 3-O-β-d-glucopyranoside with their antibacterial, anticancer, astringent, depurative and anti-inflammatory activities. A profound efforts has been done in past to address the active secondary metabolites, but still consistent struggles are required to explore the volatile compounds present in Rosa laevigata and their medicinal and therapeutic values should be investigated in future.

Effects of methanolic extract from leaves of Rubus imperialis in DSS-induced colitis in mice.

This study investigated the effects of Rubus imperialis, a berry known as "amora-branca", in colitis dextran sulfate sodium (DSS)-induced in mice. Animals were treated orally with vehicle (water), 5-aminosalicylic acid(100mg/kg) or methanolic extract from leaves of R. imperialis (MERI, 100mg/kg), once a day during seven days. The disease activity index (DAI) was observed daily. Colons were collected for histological, histochemical and biochemical analysis. The administration of MERI exacerbated colitis, as indicated by DAI heightened weight loss and increased histological colonic injury. MERI also decreased the colon mucin levels and increased colonic TNF content. The colonic levels of reduced glutathione and the superoxide dismutase activity in colitic group treated with MERI were decreased. Despite the worsening of colitis, MERI not altered the intestinal transit, body weight, colon length or organs weight in normal mice. Tormentic acid (TA) and 2β,3β,19α-trihydroxyursolic acid (THA), compounds isolated from MERI, reduced the L929 cells viability. Thus, MERI may have aggravated the DSS-induced colitis through intense intestinal mucus barrier impairment, which would lead to inflammatory responses, TA and THA contribute to the intestinal damage verified suggesting caution about the use of R. imperialis preparations, particularly in inflammatory bowel diseases.

High Triterpenic Acids Production in Callus Cultures from Fruit Pulp of Two Apple Varieties.

Very rarely fruit pulp has been used in in vitro culture to produce secondary metabolites useful in promoting health. The aims of this work were the study of the best conditions to obtain the callus cultures from the pulp of two varieties of apples, Golden Delicious (GD) and "Mela Rosa Marchigiana" (MRM), and the quali-quantitative analysis of secondary metabolites produced by the two in vitro callus cultures. Callus was induced on both Murashige and Skoog and Gamborg B5 media containing various combinations of supplements. To achieve the maximum recovery of secondary metabolites produced, preliminary extraction tests were carried out on GD apple culture using two different organic solvents (MeOH and EtOAc). The quali-quantitative analysis of the methanolic extract of both cultures was carried out by ESI-MSn and GC-MS techniques. The GC-MS analysis revealed the presence of triterpenic acids, in particular, oleanolic, ursolic, maslinic, pomolic, tormentic, corosolic and annurcoic acid along with a phytosterol, β-sitosterol. In addition, GD callus culture produced phloridzin, absent in the MRM culture. In this last culture, however, the total amount of secondary metabolites was markedly higher. The in vivo production of these bioactive compounds were also quantified in the GD and MRM apple pulps. Apple pulps produced higher amounts of triterpenic acids in vitro than in vivo. The present work can be considered a method to amplify the production of important secondary metabolites which exert beneficial effects on human health. Copyright © 2016 John Wiley & Sons, Ltd.

Megastigmane Glycosides from Docynia indica and Their Anti‐inflammatory Activities

Using various chromatographic methods, three new megastigmane glycosides, docynicasides A – C (1 – 3) and ten known, (6S,9R)‐vomifoliol 9‐O‐β‐d‐xylopyranosyl‐(1′′→6′)‐O‐β‐d‐glucopyranoside (4), hyperin (5), quercitrin (6), quercetin 3‐α‐l‐arabinofuranoside (7), naringenin 7‐O‐β‐d‐glucopyranoside (8), phloridzin (9), phloretin 2′‐O‐β‐d‐xylopyranosyl‐(1→6)‐β‐d‐glucopyranoside (10), pinosylvin 3‐O‐β‐d‐glucopyranoside (11), tormentic acid (12), and chlorogenic acid methyl ester (13) were isolated from the fruits of Docynia indica. Their chemical structures were elucidated by physical and chemical methods. All the isolated compounds were evaluated for the inhibitory activity on NO production in LPS‐stimulated BV2 cells. As the results, compounds 3 – 5 showed significant inhibitory activity on LPS‐stimulated NO production in BV2 cells with the IC50 values ranging from 21.0 to 29.3 μm.

Tormentic acid inhibits H2O2-induced oxidative stress and inflammation in rat vascular smooth muscle cells via inhibition of the NF-κB signaling pathway.

Tormentic acid (TA) is a triterpene isolated from the stem bark of the plant Vochysia divergens and has been reported to exhibit anticancer, anti-inflammatory and anti-atherogenic properties. However, the functions of TA in hydrogen peroxide (H2O2)-induced oxidative stress and inflammation in rat vascular smooth muscle cells (RVSMCs) remain unclear. Therefore, the present study aimed to investigate whether TA suppressed H2O2-induced oxidative stress and inflammation in RVSMCs, and to determine its molecular mechanisms. The present study demonstrated that TA inhibited reactive oxygen species (ROS) generation, induced H2O2 in RVSMCs, and inhibited H2O2-induced expression of inducible nitric oxide synthase (iNOS) and NADPH oxidase (NOX) in RVSMCs. In addition, TA significantly decreased the production of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6) and IL-1β. Furthermore, TA pretreatment prevented nuclear factor-κB (NF-κB) subunit p65 phosphorylation and NF-κB inhibitor α (IκBα) degradation induced by H2O2 in RVSMCs. TA is, therefore, suggested to inhibit H2O2-induced oxidative stress and inflammation in RVSMCs via inhibition of the NF-κB signaling pathway. TA may have potential as a pharmacological agent in the prevention or treatment of atherosclerosis.

Lipopolysaccharide-Induced Nitric Oxide and Prolyl Oligopeptidase – Inhibitory Activities of Triterpenoids from Sanguisorba officinalis

Lipopolysaccharide-induced nitric oxide and the protease prolyl oligopeptidase (EC 3.4.21.26) have been proposed as targets for the treatment of cognitive disturbances such as dementia. In this study, nine triterpenoids isolated from the roots of Sanguisorba officinalis, including arjunic acid (1), rosamultic acid (2), haptadienic acid (3), 1β-hydroxyeuscaphic acid (4), euscaphic acid (5), tormentic acid (6), pomolic acid (7), ursolic acid (8), and oleanolic acid (9), were tested in vitro for their inhibitory activities on lipopolysaccharide-induced nitric oxide in N9 microglia cells and on prolyl oligopeptidase. Among them, 7 exhibited the strongest inhibitory activity on lipopolysaccharide-induced nitric oxide (IC50, 21.9 µM), while 8 and 9 exhibited a significant prolyl oligopeptidase inhibitory activity (IC50 18.1 and 24.0 µM, respectively). The results could provide a clue that these triterpenoids are responsible for the reported neuro-protective effect of S. officinalis.

A remarkably simple and convergent partial synthesis of pomolic acid

Mixtures of tormentic and euscaphic acid can be used to access pomolic acid using a convergent synthetic approach. This approach constitutes at present the best and shortest route to pomolic acid.

Chemical constituents from Barringtonia racemosa

To investigate the chemical constituents from Barringtonia racemosa, twelve compounds were isolated by chromatography methods and identified as 3β-p-E-coumaroymaslinic acid (1), cis-careaborin (2), careaborin (3), maslinic acid (4), 2α, 3β, 19α-trihydroxyolean-12-ene-24, 28-dioic acid (5), 3β-p-Z-coumaroylcorosolic acid (6), corosolic acid (7), 1α, 2α, 3β, 19α-tetrahydroxyurs-12-en-28-oic acid (8), 19α-hydroxyl ursolic acid (9), 3α, 19α-dihydroxyurs-12-en-24, 28-dioic acid (10), tormentic acid (11), 3-hydroxy-7, 22-dien-ergosterol（12） by the NMR and MS data analysis. Among them, compounds 1-4,7-12 were obtained from the genus Barringtonia for the first time. All the compounds didn't show nocytotoxic activity against MCF-7 and A549 cell lines (IC₅₀>50 mg•L⁻¹).

Triterpenoids from roots of Rosa laevigata

To study the triterpenoids from the roots of Rosa laevigata. The silica gel column chromatography was used to separate the chemical constituents from the roots of Rosa laevigata Michx. HPLC was used to analyze its purity, chemical and spectroscopy methods were used to determine their structures. 12 constituents were isolated and identified as(2R, 19R)methyl 2-acetyloxy-19- hydroxyl-3-oxo-urs-12-en-28-carboxylate(1), pomonic acid (2), 18, 19-seco, 2α, 3α-dihydroxy-19-oxo-urs-11, 13(18)-dien-28-oic acid(3), swinhoeic acid (4), myrianthic acid(5), 2α, 3β, 19α-trihydroxy-24-oxo-urs-12-en-oic acid (6), tormentic acid(7), arjunic acid (8), 1β-hydroxyeuscaphic acid(9), quadranoside Ⅷ (10), alpinoside(11), rubuside B (12). Compounds 1-4, 6, 9, 11-12 were obtained from this plant for the first time. Compounds 2-4, 6, 11-12 were obtained from the genus Rosa for the first time.

Tormentic Acid Inhibits IL-1β-Induced Inflammatory Response in Human Osteoarthritic Chondrocytes.

The pro-inflammatory cytokine interleukin-1beta (IL-1β) plays critical roles in pathogenesis of osteoarthritis (OA). Tormentic acid (TA), a triterpene isolated from Rosa rugosa, has anti-inflammatory activity. However, the anti-inflammatory effect of TA on OA is still unclear. So, in the present study, we examined the effect of TA on IL-1β-induced inflammatory response in primary human OA chondrocytes. Our results demonstrated that TA significantly decreased the IL-1β-stimulated expression of matrix metalloproteinase-3 (MMP-3) and MMP-13. It also inhibited the IL-1β-induced expression of inducible nitric oxide (NO) synthase (iNOS) and cyclooxygenase-2 (COX-2), as well as the production of NO and prostaglandin E2 (PGE2) in human OA chondrocytes. Furthermore, TA greatly inhibited the IL-1β-induced NF-κB activation. In conclusion, this study is the first to demonstrate the anti-inflammatory activity of TA in human OA chondrocytes. TA significantly inhibits the IL-1β-induced inflammatory response by suppressing the NF-κB signaling pathway. Thus, TA may be a potential agent in the treatment of OA.

Plastotoma rotundifolium aerial tissue extract has antibacterial activities

Despite efforts in recent years, infectious diseases remain a worldwide major public health problem. Several infectious agents have become resistant to conventional antibiotics and there is thus urgent need to discover new antimicrobial drugs to overcome resistances. Platostoma rotundifolium (Briq.) A. J. Paton, an African plant, is mainly used to treat microbial infections in traditional Burundian medicine. From the ethyl acetate extract of the aerial parts, five pentacyclic triterpenoid acids were isolated and characterized. Based on spectral analysis, these compounds were elucidated to be 2α, 3α, 19β-trihydroxyurs-12-en-28-oic acid (that was named jeremic acid) (1), 3β-hydroxyurs-12-en-28-oic acid (ursolic acid) (2), 2α, 3β-dihydroxyurs-12-en-28-oic acid (corosolic acid) (3), 2α, 3β, 19α-trihydroxyurs-2-en-28-oic acid (tormentic acid) (4) and 19-hydroxy-2-hydroxymethyl norursa-2, 12-dien-28-oic acid (hyptadienic acid) (5). Ursolic (MIC=17.5–68μM) and corosolic (MIC=17–68μM) acids showed significant antibacterial activities against both the Gram-positive Staphylococcus aureus (methicillin-susceptible and -resistant strains) and the Gram-negative Escherichia coli, which may substantiate the use of P. rotundifolium in traditional Burundian medicine. Such hydroxylated pentacyclic triterpenoid acids could point to new antimicrobial strategies that may help overcoming the antimicrobial resistances actually observed throughout the world.

Protection against MPP+-induced neurotoxicity in SH-SY5Y cells by tormentic acid via the activation of PI3-K/Akt/GSK3β pathway

The cause of Parkinson's disease (PD) could be ascribed to the progressive and selective loss of dopaminergic neurons in the substantia nigra pars compacta, and thus molecules with neuroprotective ability may have therapeutic value against PD. In the current study, the neuroprotective effects and underlying mechanisms of tormentic acid (TA), a naturally occurring triterpene extracted from medicinal plants such as Rosa rugosa and Potentilla chinensis, were evaluated in a widely used cellular PD model in which neurotoxicity was induced by MPP+ in cultured SH-SY5Y cells. We found that TA at 1–30 μM substantially protected against MPP+-induced neurotoxicity, as evidenced by the increase in cell viability, decrease in lactate dehydrogenase release and the reduction in apoptotic nuclei. Moreover, TA effectively inhibited the elevated intracellular accumulation of reactive oxygen species as well as Bax/Bcl-2 ratio caused by MPP+. Most importantly, TA markedly reversed the inhibition of protein expression of phosphorylated Akt (Ser 473) and phosphorylated GSK3β (Ser 9) caused by MPP+. LY294002, the specific inhibitor of PI3-K, significantly abrogated the up-regulated phosphorylated Akt and phosphorylated GSK3β offered by TA, suggesting that the neuroprotection of TA was mainly dependent on the activation of PI3-K/Akt/GSK3β signaling pathway. The results taken together indicate that TA may be a potential candidate for further preclinical study aimed at the prevention and treatment of PD.

Norsesquiterpenoids and triterpenoids from strawberry cv. Falandi

Falandi is a common strawberry (Fragaria × ananassa Duch.) cultivar in southern China. Further study of the chemical constituents in Falandi fruit led to the isolation of nine norsesquiterpenoids and three triterpenoids. Falandioside D (1) and falandins A (2) and B (3) were new norsesquiterpenoids, and the others excluding tormentic acid (11) were found in strawberry for the first time. Compounds 1 and 11 exhibited potent α-glucosidase inhibitory activity with the half maximal inhibitory concentration (IC50) values of 565.0 and 27.4μM in comparison to acarbose (619.9μM). Compounds 3, 7 (blumenol C glucoside), and 11 showed cytotoxicity against human nasopharyngeal carcinoma cell line CNE1 with the IC50 values of 57.6, 56.4, and 36.0μM, respectively. Among new compounds, 1 showed 2,2′-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation scavenging capacity (IC50=36.2μM). These results suggested that non-phenolic constituents were also involved in the antidiabetic, antitumour, and antioxidant effects of strawberry fruit.

Phytotoxic and Nematicidal Components of Lavandula luisieri.

Several preparations were obtained from the aerial parts of predomesticated Lavandula luisieri, including the essential oil and ethanolic, hexane, and ethyl acetate extractives. Additionally, pilot plant vapor pressure extraction was carried out at a pressure range of 0.5-1.0 bar to give a vapor pressure oil and an aqueous residue. A chemical study of the hexane extract led to the isolation of six necrodane derivatives (1, 2, and 4-7), with four of these (1, 2, 5, and 7) being new, as well as camphor, a cadinane sesquiterpene (9), tormentic acid, and ursolic acid. The EtOAc and EtOH extracts contained a mixture of phenolic compounds with rosmarinic acid being the major component. Workup of the aqueous residue resulted in the isolation of the necrodane 3 and (1R*,2S*,4R*)-p-menth-5-ene-1,2,8-triol (8), both new natural compounds. The structures of the new compounds were established based on their spectroscopic data. The phytotoxic and nematicidal activities of these compounds were evaluated.

Determination and Identification of a Specific Marker Compound for Discriminating Shrub Chaste Tree Fruit from Agnus Castus Fruit Based on LC/MS Metabolic Analysis.

Shrub Chaste Tree Fruit (SCTF) is defined as the fruits of Vitex rotundifolia L. f. and V. trifolia L. and has been used as a component of some traditional Japanese medicines (Kampo formulations). Agnus Castus Fruit (ACF) is defined as the dried ripe fruits of V. agnus-castus L.; it is used in traditional European medicines, but is becoming popular in Japan as both an over-the-counter drug and as an ingredient in health foods for treating premenstrual syndrome (PMS). To ensure the efficacy and safety of both SCTF and ACF products, it is important to precisely authenticate their botanical origins and to clearly distinguish between SCTF and ACF. Therefore, we tried to identify SCTF-specific marker compounds based on LC/MS metabolic analysis. The multivariate analysis of LC/MS data from SCTF and ACF samples furnished candidate marker compounds of SCTF. An SCTF-specific marker was isolated from SCTF crude drugs and identified as 3-O-trans-feruloyl tormentic acid on the basis of spectroscopic data from NMR and MS. Since avoiding contamination from closely related species is a significant requirement for pharmaceuticals of natural origin, this information will be valuable for the quality control of both SCTF and ACF products from the viewpoint of regulatory science.

Tormentic acid in foods exerts anti-proliferation efficacy through inducing apoptosis and cell cycle arrest

Tormentic acid (TA) is a triterpenoid saponin widely distributed in natural plant foods with various bioactivities. However, the underlying pathways through which TA exerts the bioactivities remain unclear. Here, experiments were carried out to elucidate the molecular mechanisms accounting for the antiproliferation of TA using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay, flow cytometry, quantitative real-time polymerase chain reaction and Western blotting analysis. TA exhibited a dose-dependent inhibitory effect on MCF-7 cells, which was caused by apoptosis and G0/G1 phase arrest. Reactive oxygen species (ROS) acts as an upstream mediator in TA-induced MCF-7 cell growth inhibition and intracellular ROS levels induced mitochondrial dysfunction, thereby resulting in the release of apoptogenic factors into cytosol, and then activating caspase-3 and caspase-9. TA induced cell cycle arrest via changing the cyclin D1 and cyclin-dependent kinase 4 messenger ribonucleic acid expression levels. Moreover, TA significantly down-regulated the NF-kappa-B cell survival pathway and the expression level of phosphorylated ERK.

Inhibitory effects of Ligustrum robustum (Rxob.) Blume extract on α-amylase and α-glucosidase

The α-amylase and α-glucosidase inhibitory effects of Ligustrum robustum (Rxob.) Blume extract (LRE) were investigated and the primary bioactive components were isolated and identified. With high contents of phenols and flavonoids, LRE exhibited remarkable inhibitory effects on α-amylase and α-glucosidase. It also displayed strong antioxidant activity for scavenging free radicals (superoxide and hydroxyl radicals), inhibiting lipid peroxidation and providing reducing power. Moreover, LRE effectively inhibited the catalytic activity of sucrase and maltase in Caco-2 cell monolayer. According to the bioassay-guided isolation and analyses using high-performance liquid chromatography, time-of-flight mass spectrometry, and nuclear magnetic resonance spectroscopy, pomolic acid, tormentic acid, 3-O-cis-p-coumaroyl tormentic acid and 3-O-trans-p-coumaroyl tormentic acid were characterized as the main active constituents. These results suggest that Ligustrum robustum (Rxob.) Blume with a great potential to control postprandial hyperglycaemia might be a novel resource for use in the functional food industry.