Effects Of Zerumbone Research Articles

Zerumbone exhibits anti-inflammatory effects by suppressing eicosanoid signaling: Evidence from LPS-induced peripheral blood leukocytes

Zerumbone, a sesquiterpene isolated from Zingiber zerumbet, has many bioactivities, exhibiting anti-inflammatory properties. However, the effect of zerumbone on the eicosanoid signaling pathway has yet to be examined. Here, we deciphered the anti-eicosanoid properties of zerumbone isolated from ginger. The molecular interaction between zerumbone and eicosanoid metabolizing enzymes (COX-2, 5-LOX, FLAP, and LTA4-hydrolase) and receptors (EP-4, BLT-1, and ICAM-1) along with NOS-2 were assessed using Auto-Dock 4.2 and visualized by chimera and Liggplot+ software. Further, the leukocytes were treated with zerumbone (1–20 μM) and activated using bacterial lipopolysaccharide (LPS-10 nM). The oxidative stress (OS) markers, antioxidant enzymes, and the eicosanoid pathway mediators such as COX-2, 5-LOX, BLT-1, and EP-4 were assessed. The molecular interaction of zerumbone with eicosanoids showed a higher binding affinity with mPGES-1, followed by NOS-2, FLAP, COX-2, LTA-4-hydrolase, and BLT-1. The concentration of 5 μM zerumbone effectively prevented the generation of reactive oxygen species (ROS) and nitric oxide (NO). Likewise, zerumbone significantly (p<0.05) inhibited COX-2, 5-LOX, NOS-2, EP-4, BLT-1, and ICAM-1 expression in LPS-induced peripheral blood leukocytes from rats. Further, the zerumbone treatment on the human PBMCs activated with LPS showed significant inhibition in the expression of ICAM1, COX-2, 5-LOX, and the generation of inflammatory cytokines compared to the control. Overall, the data presented infers that zerumbone positively modulates critical enzymes and receptors of eicosanoids in leukocytes activated with lipopolysaccharides. Thus, zerumbone can be a potential anti-eicosanoid drug in managing inflammation.

Dietary bitter ginger-derived zerumbone improved memory performance during aging through inhibition of the PERK/CHOP-dependent endoplasmic reticulum stress pathway.

PERK/CHOP pathway-mediated excessive endoplasmic reticulum (ER) stress is closely linked to aging-related cognitive impairment (ARCD). Zerumbone (ZB), a naturally occurring sesquiterpene molecule obtained from dietary bitter ginger, has garnered significant interest due to its diverse range of biological properties. It is unclear, though, if ZB can reduce ARCD by preventing ER stress that is dependent on the PERK/CHOP pathway. Here, the PERK-CHOP ER stress pathway was the main focus of an evaluation of the effects and mechanisms of ZB for attenuating ARCD in D-galactose (D-gal)-induced aging mice and SH-SY5Y cells. According to our findings, ZB not only greatly decreased neuronal impairment both in vitro and in vivo, but also significantly alleviated learning and memory failure in vivo. ZB significantly reduced the activation of the PERK/CHOP pathway and neuronal apoptosis in vitro and in vivo, exhibiting the down-regulation of GRP78, p-PREK/PERK, and CHOP expression levels, in addition to suppressing oxidative damage (MDA drop and SOD rise). Comparable outcomes were noted in SH-SY5Y cells subjected to severe ER stress caused by TM. On the other hand, 4-PBA, an ER stress inhibitor, considerably reversed these modifications. Remarkably, CCT020312 (a PERK activator) dramatically overrode the inhibitory effects of ZB on the PERK/CHOP pathway and neuronal death in D-gal-induced SH-SY5Y cells. In contrast, GSK2606414 (a PERK inhibitor) significantly increased these effects of ZB. In summary, our results suggested that ZB prevented D-gal-induced cognitive deficits by blocking the PERK/CHOP-dependent ER stress pathway and apoptosis, suggesting that ZB might be a natural sesquiterpene molecule that relieves ARCD.

Phytochemical mediated modulation of COX-3 and NFκB for the management and treatment of arthritis

In this study, we investigated whether zerumbone (ZBN), ellagic acid (ELA) and quercetin (QCT), the plant-derived components, can modulate the role of COX-3 or cytokines liable in arthritic disorder. Initially, the effect of ZBN, ELA, and QCT on inflammatory process was investigated using in-vitro models. In-silico docking and molecular dynamics study of these molecules with respective targets also corroborate with in-vitro studies. Further, the in-vivo anti-arthritic potential of these molecules in Complete Freund’s adjuvant (CFA)-induced arthritic rats was confirmed. CFA increases in TNF-α and IL-1β levels in the arthritic control animals were significantly (***p < 0.001) attenuated in the ZBN- and ELA-treated animals. CFA-induced attenuation in IL-10 levels recovered under treatment. Moreover, ELA attenuated CFA-induced upregulation of COX-3 and ZBN downregulated CFA-triggered NFκB expression in arthritic animals. The bonding patterns of zerumbone in the catalytic sites of targets provide a useful hint in designing and developing suitable derivatives that can be used as a potential drug. To our best knowledge, the first time we are reporting the role of COX-3 in the treatment of arthritic disorders which could provide a novel therapeutic approach for the treatment of inflammatory disorders.

Zerumbone suppresses high glucose and LPS-induced inflammation in THP-1-derived macrophages by inhibiting the NF-κB/TLR signaling pathway

Diabetes mellitus is characterized by hyperglycemia. Low-grade bacterial infection with hyperglycemia in patients with diabetes is associated with atherosclerosis development. Therefore, this study hypothesized that macrophages lead to more severe diabetic complications under combined conditions of high glucose and lipopolysaccharide (LPS)-induced inflammation than under normoglycemic conditions. Zerumbone is the main component of Zingiber zerumbet Smith essential oil, a type of wild ginger. It possesses various biomedical activities, including antibacterial, antioxidant, anti-inflammatory, and anticancer activities; however, the precise mechanism of its anti-inflammatory and epigenetic effects is not fully understood. In this study, the effects of zerumbone on the secretion of proinflammatory cytokines and its underlying regulatory mechanism were investigated in THP-1-derived macrophages exposed to high glucose and LPS. THP-1-derived macrophages were cultured under normoglycemic (5.5 mmol/L glucose) or hyperglycemic (25 mmol/L glucose) conditions in the absence or presence of zerumbone (5-50 μM) for 48 hours and then treated with 100 ng/mL LPS for 6 hours. Zerumbone (25 and 50 μM) suppressed the production of tumor necrosis factor-α and interleukin-6 and the activation of cyclooxygenase-2, nuclear factor-κB, histone deacetylases 3 proteins, and Toll-like receptor messenger RNA (mRNA) and increased the transcription of the sirtuin 1 (SIRT1), SIRT3, and SIRT6 mRNAs. Taken together, our results suggest that zerumbone may exert beneficial effects on diabetes and its complications.

Abstract 9508: The Natural Product Zerumbone Suppresses Pressure Overload-Induced Cardiac Dysfunction by Inhibiting Cardiac Hypertrophy and Fibrosis

Introduction: Cardiac hypertrophy and fibrosis are hallmarks of cardiac remodeling and are involved functionally in the development of heart failure (HF). Various food components effectively prevent cardiovascular diseases. However, it is unknown whether zerumbone (ZER), a major active terpene found in endemic wild ginger species, suppresses cardiac hypertrophy, fibrosis, and dysfunction. Hypothesis: We assessed the hypothesis that ZER suppresses cardiac hypertrophy and fibrosis in vitro and in vivo . Methods: The effects of ZER on phenylephrine (PE)-induced hypertrophic responses and on transforming growth factor beta (TGF-β)-induced fibrotic responses were examined in primary cultured cardiomyocytes and fibroblasts from neonatal rats. To determine whether ZER prevents the development of pressure overload-induced HF in vivo , a transverse aortic constriction mouse model was utilized (n=6-10). One day after surgery, mice were randomly divided into two groups and orally administered with ZER 20 mg / kg or vehicle for 8 weeks. Cardiac function was evaluated by echocardiography. Changes in cardiomyocyte surface area and degree of fibrosis were observed by histological analysis (HE and WGA staining). The total mRNA levels of the genes associated with hypertrophy and fibrosis were measured by qRT-PCR. Protein expression (Akt phosphorylation and α-SMA) were assessed by western blotting. Results: ZER significantly suppressed PE-induced increases in cell size, hypertrophic gene expression (ANF and BNP), and Akt phosphorylation in cardiomyocytes. TGF-β-induced increases in collagen synthesis, mRNA levels of POSTN and α-SMA, and protein expression of α-SMA were lower in the ZER-treated cultured cardiac fibroblasts. Echocardiography results showed that left ventricular fractional shortening was increased and wall thickness was reduced in the ZER group compared with the vehicle group. Histological analysis showed that pressure overload-induced cardiac hypertrophy and cardiac fibrosis were inhibited in the ZER group compared with the vehicle group. Conclusions: These results suggest that zerumbone ameliorates pressure overload-induced cardiac dysfunction, at least in part by suppressing both cardiac hypertrophy and fibrosis.

Zerumbone attenuates house dust mite extract-induced epithelial barrier dysfunction in 16HBE14o- cells

Context The airway epithelial barrier can be disrupted by house dust mite (HDM) allergens leading to allergic airway inflammation. Zerumbone, a natural monocyclic sesquiterpene, was previously found to possess anti-asthmatic effect by modulating Th1/Th2 cytokines. However, the protective role of zerumbone on epithelial barrier function remains to be fully explored. Objective To investigate the effect of zerumbone on HDM extract-induced airway epithelial barrier dysfunction. Materials and methods Human bronchial epithelial cells 16HBE14o- were incubated with 100 μg/mL HDM extract and treated with non-cytotoxic concentrations of zerumbone (6.25 μM, 12.5 μM, and 25 μM) for 24 h. The epithelial junctional integrity and permeability were evaluated through transepithelial electrical resistance (TEER) and fluorescein isothiocynate (FITC)-Dextran permeability assays, respectively. The localization of junctional proteins, occludin and zona occludens (ZO)-1, was studied using immunofluorescence (IF) while the protein expression was measured by western blot. Results Zerumbone inhibited changes in junctional integrity (6.25 μM, p ≤ .05; 12.5 μM, p ≤ .001; 25 μM, p ≤ .001) and permeability (6.25 μM, p ≤ .05; 12.5 μM, p ≤ .01; 25 μM, p ≤ .001) triggered by HDM extract in a concentration-dependent manner. This protective effect could be explained by the preservation of occludin (12.5 μM, p ≤ .01 and 25 μM, p ≤ .001) and ZO-1 (12.5 μM, p ≤ .05 and 25 μM, p ≤ .001) localization, rather than the prevention of their cleavage. Discussion and conclusion Zerumbone attenuates HDM extract-induced epithelial barrier dysfunction which supports its potential application for the treatment of inflammation-driven airway diseases such as asthma.

The Potential Anti-Inflammatory Effects of Zerumbone in COVID-19 Patients

The article's abstract is not available.

The Combination of Zerumbone and 5-FU: A Significant Therapeutic Strategy in Sensitizing Colorectal Cancer Cells to Treatment

Objectives. Chemotherapy is considered to be essential in the treatment of patients with colorectal cancer (CRC), but drug resistance reduces its efficacy. Many patients with advanced CRC eventually show resistance to 5-fluorouracil (5-FU) therapy. Synergistic and potentiating effects of combination therapy, using herbal and chemical drugs, can improve patients’ response. Zerumbone (ZER), which is derived from ginger, has been studied for its growth inhibitory function in various types of cancer. Methods. The cytotoxic effects of ZER and 5-FU alone and their combination, on the SW48 and HCT-116 cells, were examined, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). The mRNA and protein levels of β-catenin, survivin, and vimentin were measured in treated CRC cells, using qRT-PCR and western blot. Colony formation assay, scratch test, and flow cytometry were performed to detect the changes of proliferation, migration, and apoptosis. Key Findings. In HCT-116- and SW48-treated cells, the proliferation, the gene and protein expression levels of the markers, the migration, the colony formation, and the survival rates were all significantly reduced compared to the control groups, and the sharpest decline was observed in the 5-FU+ZER treatment groups. Conclusions. Combination therapy has shown promising results in CRC cells, especially in drug-resistant cells.

Computational Investigation of Zerumbone as an Inhibitor of TNF-alpha Using Molecular Dynamics and Molecular Docking Methods

Background: There are several reports on the anti-cancer property of zerumbone, such as in breast, cervical and ovarian cancer. But the investigation of the actual protein target has been the least concerned and there are few reports on the inhibitory effect of zerumbone against specific cancer-causing proteins and enzymes. Therefore, further investigation is required at a much deeper molecular level. Objective: This study aimed to determine the anti-proliferative activity of Zerumbone against cervical cancer cell and assess its TNF-α enzyme inhibitory action. Methods: The investigation emphasized the anti-cancer activity of zerumbone against HeLa cells and its subsequent TNF-α assay. Furthermore, computational studies on Zerumbone as an inhibitor of TNF-α were carried out using computational techniques such as docking and MD simulations. Results and Discussion: From the molecular docking analysis, it was observed and substantiated that the α,β-Unsaturated carbonyl scaffold is the main driving force for anti-cancer activity of zerumbone and inhibition of TNF-α. Conclusion: Zerumbone might be a potent anti-cancer agent targeting the HeLa cancer cell lines and inhibiting the TNF-α enzyme.

Involvement of Wnt/β-catenin pathway in the inhibition of invasion and epithelial-mesenchymal transition in ovarian cancer cells

Purpose: To investigate the effects of zerumbone on cell invasion, epithelial-mesenchymal transition (EMT) and the potential signaling pathway involved in ovarian cancer cells.Methods: Caov-3 cell proliferation was assessed using 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-diphenytetrazoliumromide (MTT) assay. Wound healing assay was used to determine Caov-3 cell migration while cell invasion was evaluated using Transwell assay. Protein expression was determinedby western blot.Results: Cell viability was reduced by 5, 10, 20, and 50 μM zerumbone (p &lt; 0.05) in a concentrationdependent manner while cell migration and invasion were inhibited by 10 and 20 μM zerumbone (p &lt; 0.05). Protein expression levels of E-cadherin and cytoplasm β-catenin were upregulated by zerumbone (p &lt; 0.05) in a concentration-dependent manner. On the other hand, protein expression levels of Ncadherin, vimentin, ZEB1, nuclear β-catenin, and c-Myc were suppressed by zerumbone (p &lt; 0.05) also in a concentration-dependent manner.Conclusion: The results demonstrate that zerumbone inhibits cell proliferation, migration and invasion, but represses the EMT process via inactivation of Wnt/β-catenin signaling pathway.&#x0D; Keywords: Zerumbone, Ovarian cancer, Wnt/β-catenin pathway, Epithelial-mesenchymal transition

Zerumbone decreases BACH1 levels by upregulating miR- 708 to inhibit breast cancer cell proliferation and invasion

Purpose: To investigate the potential mechanism by which zerumbone suppresses breast cancer (BC) cells.Methods: Cell viability and Transwell assays were performed to assess the effect of zerumbone on BC cell growth. The downstream target of zerumbone was determined using quantitative polymerase chain reaction assays and immunoblotting. Cell viability assays and immunoblotting were conducted to detect if zerumbone had any effect on BACH1 (BTB domain and CNC homolog 1) expression.Results: Zerumbone suppressed the proliferation, migration, and invasion of BC cells. It also upregulated the expression of microRNA (miR)-708 and, hence, suppressed BACH1 expression. Furthermore, zerumbone suppressed the proliferation and invasion of BC cells by promoting miR-708expression and suppressing BACH1.Conclusion: The findings help clarify the anti-tumor mechanism of zerumbone and provide theoretical and therapeutic bases for the anti-tumor effects of Chinese herbal medicine.&#x0D; Keywords: Breast cancer, Zerumbone, Cell invasion, MiR-708, BACH1

Zerumbone attenuates lipopolysaccharide-induced activation of BV-2 microglial cells via NF-\u03baB signaling

The brain is considered an immune-privileged organ. However, it has been found that inflammation mediated by microglia, which were once believed to support the brain structure, plays important roles in neuronal cell survival and death. Whether activated microglia has beneficial or detrimental effects on neurons remain controversial. Activated microglia could contribute to maintaining homeostasis in the brain by removing damaged cells. Nonetheless, dysregulation of microglial activation leads to neuronal cell death. Therefore, much attention has been paid to compounds that regulate microglial activation. Zerumbone, a constituent of Zingiber zerumbet, has been reported to exert several biological activities such as anticancer, anti-bacterial, and anti-inflammatory effects. In this study, we aimed to determine the anti-inflammatory effect of zerumbone on lipopolysaccharide-induced activation of BV-2 microglial cells and elucidate the underlying mechanism of action. Zerumbone suppressed nitric oxide and prostaglandin E2 production induced by lipopolysaccharides through inhibiting the expression of inducible nitric oxide synthase and cyclooxygenase-2. Blocking of mitogen-activated protein kinase and NF-κB activation, if not completely, is considered to be due to the anti-inflammatory effect of zerumbone against microglial activation.

Zerumbone Reduces the Level of MAPK1 by Upregulating Mir-758-3p to Inhibit the Survival of Thyroid Papillary Cancer Cells

Thyroid papillary carcinoma is the most common type of thyroid malignancy. The mitogen-activated protein kinase signaling cascade is the major pathway involved in thyroid papillary carcinoma and may be regulated by small molecules and transcription factors. Zerumbone is a bioactive compound with multiple pharmacological properties. This study investigated the effects of zerumbone on survival and proliferation of thyroid papillary carcinoma-1 cells with an emphasis on the level of microRNA-758-3p and its target gene, mitogen-activated protein kinase-1. Zerumbone exhibited dose-dependent inhibitory effects on the viability and proliferation of thyroid papillary carcinoma-1 cells. It also gradually elevated the level of microRNA-758-3p but decreased mitogen-activated protein kinase-1 expression in these cells. Mitogen-activated protein kinase-1 has been suggested as a downstream target of microRNA-758-3p and confirmed by dual-reporter luciferase assay. The downregulation of microRNA-758-3p in thyroid papillary carcinoma-1 cells via the transfection with microRNA-758-3p inhibitor reversed the inhibitory effect of zerumbone on mitogen-activated protein kinase-1 expression. Furthermore, the transfection of thyroid papillary carcinoma-1 cells with lentiviral vector overexpressing mitogen-activated protein kinase-1 eliminated the protective effects of zerumbone on thyroid papillary carcinoma progression as compared to the group delivered with control vector. In conclusion, this study demonstrated that zerumbone inhibited survival and proliferation of thyroid papillary carcinoma-1 cells via suppressing the expression of mitogen-activated protein kinase-1 through microRNA-758-3p. These data suggest the therapeutic potential of zerumbone in thyroid papillary carcinoma treatment.

Zerumbone ameliorates behavioral impairments and neuropathology in transgenic APP/PS1 mice by suppressing MAPK signaling

BackgroundAlzheimer’s disease (AD) is a major clinical problem, but there is a distinct lack of effective therapeutic drugs for this disease. We investigated the potential therapeutic effects of zerumbone, a subtropical ginger sesquiterpene, in transgenic APP/PS1 mice, rodent models of AD which exhibit cerebral amyloidosis and neuroinflammation.MethodsThe N9 microglial cell line and primary microglial cells were cultured to investigate the effects of zerumbone on microglia. APP/PS1 mice were treated with zerumbone, and non-cognitive and cognitive behavioral impairments were assessed and compared between the treatment and control groups. The animals were then sacrificed, and tissues were collected for further analysis. The potential therapeutic mechanism of zerumbone and the signaling pathways involved were also investigated by RT-PCR, western blot, nitric oxide detection, enzyme-linked immunosorbent assay, immunohistochemistry, immunofluorescence, and flow cytometry analysis.ResultsZerumbone suppressed the expression of pro-inflammatory cytokines and induced a switch in microglial phenotype from the classic inflammatory phenotype to the alternative anti-inflammatory phenotype by inhibiting the mitogen-activated protein kinase (MAPK)/nuclear factor-kappa B signaling pathway in vitro. After a treatment period of 20 days, zerumbone significantly ameliorated deficits in both non-cognitive and cognitive behaviors in transgenic APP/PS1 mice. Zerumbone significantly reduced β-amyloid deposition and attenuated pro-inflammatory microglial activation in the cortex and hippocampus. Interestingly, zerumbone significantly increased the proportion of anti-inflammatory microglia among all activated microglia, potentially contributing to reduced β-amyloid deposition by enhancing phagocytosis. Meanwhile, zerumbone also reduced the expression of key molecules of the MAPK pathway, such as p38 and extracellular signal-regulated kinase.ConclusionsOverall, zerumbone effectively ameliorated behavioral impairments, attenuated neuroinflammation, and reduced β-amyloid deposition in transgenic APP/PS1 mice. Zerumbone exhibited substantial anti-inflammatory activity in microglial cells and induced a phenotypic switch in microglia from the pro-inflammatory phenotype to the anti-inflammatory phenotype by inhibiting the MAPK signaling pathway, which may play an important role in its neuroprotective effects. Our results suggest that zerumbone is a potential therapeutic agent for human neuroinflammatory and neurodegenerative diseases, in particular AD.

Effects of zerumbone on proliferation and apoptosis of esophageal cancer cells and on P53 and Bcl-2 expression levels.

The effects of zerumbone on the proliferation and apoptosis of esophagus cancer cells and on the P53 and Bcl-2 expression levels were studied. The esophagus cancer EC-109 cells were cultured and inoculated. The effect of zerumbone on proliferation of EC-109 cells was detected via the Cell Counting Kit-8 (CCK-8) method. Cell apoptosis was detected via TdT-mediated dUTP nick end-labeling (TUNEL) staining. Moreover, the mRNA expression levels of P53 and Bcl-2 were detected via reverse transcription-polymerase chain reaction (RT-PCR), and the protein expression levels of P53 and Bcl-2 were evaluated via western blotting. CCK-8 detection results showed that compared with control group, zerumbone in different concentrations could inhibit the activity of EC-109, and the proliferation inhibition rate was significantly increased in a concentration-dependent manner with the increase of concentration. TUNEL staining showed that cell apoptosis gradually occurred in administration group, and the number of apoptotic cells was increased in a concentration-dependent manner with the increase of concentration. RT-PCR detection results showed that the mRNA expression level of P53 in administration group was significantly increased compared with that in control group, but that of Bcl-2 was significantly decreased. Western blotting showed that the protein expression level of Bcl-2 in administration group in different concentrations was significantly increased with the increase of zerumbone concentration, but that of Bcl-2 was significantly decreased in a concentration-dependent manner. Zerumbone can inhibit the proliferation and induce apoptosis of esophageal cancer EC-109 cells, and its induction of apoptosis may be realized through upregulating the mRNA expression of P53 and downregulating the mRNA expression of Bcl-2, and upregulating the protein expression of P53 and downregulating the protein expression of Bcl-2.

Retracted: Zerumbone inhibits epithelial‐mesenchymal transition and cancer stem cells properties by inhibiting the β‐catenin pathway through miR‐200c

Colorectal cancer (CRC) is one of the most lethal and rampant human malignancies in the world. Zerumbone, a sesquiterpene isolated from subtropical ginger, has been found to exhibit an antitumor effect in various cancer types. However, the effect of Zerumbone on the biological properties of CRC, including epithelial-mesenchymal transition (EMT) and cancer stem cells (CSCs) has not been fully elucidated. Here, we investigated the inhibitory action of Zerumbone on the EMT process, CSC markers, and the β-catenin signaling pathway in the presence or absence of miR-200c. The effect of Zerumbone on HCT-116 and SW-48 cells viability was examined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. The effects of Zerumbone on EMT-related genes, CSCs markers, cell migration, invasion, sphere-forming, andβ-catenin signaling pathway were explored. To evaluate the role of miR-200c in anticancer effects by Zerumbone, miR-200c was downregulated by LNA-anti-miR-200c. Zerumbone significantly inhibited cell viability, migration, invasion, and sphere-forming potential in HCT-116 and SW-48 cell lines. Zerumbone significantly suppressed the EMT and CSC properties as well as downregulated the β-catenin. Silencing of miR200c reduced the inhibitory effects of Zerumbone on EMT and CSCs in CRC cells. These data indicated that Zerumbone may be a promising candidate for reducing the risk of CRC progression by suppressing the β-catenin pathway via miR-200c.

In Vitro Genotoxic Effects of Zerumbone and Cisplatin Combination in CHO Cell Lines

Objective: Several natural products are being increasingly used in the treatment of cancer to minimize the adverse side effects of cancer chemotherapy. Zerumbone (ZER), the sesquiterpene derived from Zingiber zerumbet Smith, has been reported to have an in vitro anticancer effects against various human tumour cells as well as in vivo against a number of induced malignancies in mice. Previously we have reported the genotoxic effects of ZER in vitro against CHO cell lines. Material and Method: The aim of this study was to investigate the genotoxic effects of the combination of ZER along with cisplatin in CHO cells. Two cytogenetic endpoints were used, namely Chromosomal Aberrations assay (CA) and Micronucleus test (MN). Both cytogenetic endpoints were performed without any metabolic activation. Result: ZER treated cultures showed the significant increase in the frequency of the chromosome aberrations and MN induction. In CA assay, marked changes have been observed after co-treatment of CHO cell lines with different concentration of ZER along with 5 µM Cisplatin when compared to ZER treatment alone, suggesting a possible synergistic genetoxic effects. Whereas, treatment of CHO cell lines with different concentrations of ZER along with 2.5 µM Cisplatin was found to reduce chromosomal aberrations, suggesting an antagonistic genotoxic effect. On the other hand, in MN induction test, co-treatment of CHO cell lines with both 2.5 µM and 5 µM Cisplatin and different concentration of ZER found to reduce the genotoxic effects compared to the 2.5 µM and 5 µM Cisplatin alone suggesting an antagonistic genotoxic effect.Conclusion: The genotoxic effects of combined low concentrations of Cisplatin with different concentrations of ZER could have an antagonist genotoxic potential in vitro in CHO cell lines.

Cytotoxic and Genotoxic Effects of Zerumbone on WEHI 7.2 Wild Type Murine Thymoma Cells

Zerumbone (ZER), a sesquiterpene in the rhizomes of Zingiber zerumbet Smith, was shown to exhibit antiproliferative activities on various cancer cells. This study was carried out to determine the cytotoxic and genotoxic effects of ZER on WEHI 7.2 wild type murine thymoma cells through the employment of standard MTT assay, alkaline comet assay and flow cytometry Annexin V/PI. Results from the MTT assay demonstrated that ZER has a dose-dependent but not a time-dependent cytotoxic effect towards WEHI 7.2 wild type cells with IC50 values at 24, 48 and 72 hours were 3.02±0.20 µg/ml (13.832 µM), 2.73±0.13 µg/ml (12.503 µM) and 2.65±0.13 µg/ml (12.137 µM) respectively. Using IC10 and IC25 values obtained from the MTT assay, alkaline comet assay was carried out to detect DNA damage in ZER treated cells at three different time points (1/2 h, 1 h and 2 h). From the results, it was found that ZER induced significant DNA damage at all three time points for both concentrations (p &lt; 0.05). Comparison of DNA damage levels at both concentrations suggested a concentration-dependent genotoxicity, as significantly higher values of tail DNA percentage and tail moment were obtained for cells treated with IC25 concentration (p &lt; 0.05). Furthermore, to understand the mode of cell death induced by ZER, flow cytometry Annexin V/PI was performed and it was found that cytotoxicity was achieved primarily via apoptosis. Collectively, ZER is able to induce genotoxicity in treated cells which subsequently leads to cytotoxicity via apoptosis and these presented characteristics suggest the compound as a potential anticancer drug.

Anti‐inflammatory Effects of Zerumbone on Chronic Inflammation Models in Differentiated Human THP‐1 Monocytes Induced by Lipopolysaccharide

Inflammation plays a key role in the pathogenesis of various human diseases, such as cancer, chronic asthma, rheumatoid arthritis, and psoriasis. In analyses of pathogen‐associated molecular patterns (PAMPs), at least four families of mammalian innate immune receptors, known as pattern recognition receptors (PRPs), have been identified, including toll‐like receptor (TLR). TLR plays a critical role in triggering immune responses by sensing conserved microbial‐derived macromolecules. Lipopolysaccharide (LPS), a product of gram‐negative bacteria, stimulates the TLR4 complex, particularly in macrophages and dendritic cells. LPS plays a vital role not only in leading inflammatory processes but also in stimulating the production of inflammatory mediators. For these reasons, LPS‐activated THP‐1 macrophages have been used as in vitro models for evaluating anti‐inflammatory effects. Zerumbone is the main component of the tropical plant Zingiber zerumbet Smith and has been reported to exert anti‐inflammatory, anti‐ulcer, and anti‐hyperglycemic effects. However, the specific mechanisms through which zerumbone exerts these anti‐inflammatory effects in the context of LPS‐induced inflammation in differentiated human THP‐1 monocytes is not fully understood. THP‐1 cells were differentiated into macrophages by phorbol 12‐myristate 13‐acetate (PMA, 1 μM) for 48 h. Differentiated THP‐1 cells were cultured with or without zerumbone (1–10 μM) for 48 h and then treated with 500 ng/mL LPS 6 h prior to harvesting. In this study, we found that zerumbone suppressed the secretion of pro‐inflammatory markers (cyclooxygenase‐2, interleukin‐6, and tumor necrosis factor‐α) and expression of nuclear factor‐κB in LPS‐stimulated THP‐1 macrophages. Histone deacetylases (HDACs) remove the acetyl group from lysine residues within a range of proteins, including transcription factors and histones. Thus, we examined HDAC3 and SIRT1 activity in differentiated THP‐1 cells. Although LPS treatment reduced or induced HDAC3 expression, zerumbone increased SIRT1 and suppressed HDAC3, as demonstrating using enzyme‐linked immunosorbent assay, real‐time polymerase chain reaction, western blotting, and immunofluorescence analysis. H3K9ac, which is regulated by histone acetylating enzymes, has an open chromatin structure that activates gene expression. The expression of H3K9ac was decreased following LPS stimulation plus zerumbone treatment. Zerumbone treatment significantly inhibited Toll‐like receptor 2/4 protein and mRNA expression and MyD88 gene expression under LPS‐induced THP‐1 macrophages. Thus, based on the results of this study, zerumbone may be a promising candidate for the treatment and prevention of chronic inflammatory diseases.

Essential oil composition, antifungal, and seedling growth inhibitory effects of zerumbone from Zingiber zerumbet Smith

The composition of the essential oil isolated from the rhizomes of Zingiber zerumbet Smith was determined using GC and GC–MS. Zerumbone (74.82%), humulene (6.02%), and β-copaen-4α-ol (4.32%) were identified as major compounds. E-Bisabol-11-ol (4.32%), E-bisabol-11-ol (0.92%), β-sinensal (0.57%), and α-himachalene oxide (0.19) were reported for the first time in its oil. Zerumbone was further isolated from the crude mixture of oil and identified by FTIR, NMR (1H, 13C, DEPT, HXCO, and HMBC), and mass (EI-MS) spectral data. Zerumbone showed promising antifungal activity against three phytopathogenic fungi, namely Rhizoctonia solani (EC50, 39.6 ppm), Sclerotium rolfsii (EC50, 59.3 ppm), and Macrophomina phaseolina (EC50, 147.4 ppm) compared with hexaconazole (EC50, 18.3, 13.4, and 4.5 ppm, respectively). Zerumbone has also shown concentration-dependent affect on seedling growth of Phalaris minor Retz., compared with control and Triticum aestivum L. (wheat). Zerumbone strongly suppressed the root and shoot growth of P. minor seedling at 1000 ppm compared with control and wheat. It exhibited no or less effect on the germination of seeds of T. aestivum.