The dual-targeting mechanism of an anti-inflammatory diarylheptanoid from Curcuma zedoaria (Christm.) Roscoe with the capacity for ß2-adrenoreceptor agonism and NLRP3 inhibition.

Chronic obstructive pulmonary disease (COPD) is a common respiratory disease characterized by symptoms of shortness of breath and chronic inflammation. Curcuma zedoaria (Christm.) Roscoe is a well-documented traditional medical herb that is frequently used in the treatment of COPD. Previously, we identified a diarylheptanoid compound (1-(4-hydroxy-5-methoxyphenyl)-7-(4,5-dihydroxyphenyl)-3,5-dihydroxyheptane; abbreviated as HMDD) from this herb that exhibited potent agonistic activity on ß2-adrenergic receptors (ß2 adrenoreceptor) that are present on airway smooth muscle cells. In this work, we used chemically synthesized HMDD compound, and confirmed its bioactivity on ß2 adrenoreceptors. Then by a proteomics study and anti-inflammatory evaluation detections, we found that HMDD downregulated the nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) signaling pathway and suppressed the NLRP3 receptor expression in RAW264.7 macrophages and in a COPD model in A549 lung carcinoma cells. HMDD also decreased nitric oxide production levels, and impacted other interleukins and the phosphorylation of NF-κB and ERK pathways. We performed molecular docking of HMDD on ß2 adrenoreceptor and NLRP3 protein models. This work reports the anti-inflammatory effects of HMDD and suggests a dual-targeting mechanism of ß2-adrenoreceptor agonism and NLRP3 inhibition. Such a mechanism scientifically supports the clinical uses of Curcuma zedoaria (Christm.) Roscoe in treating COPD, as it can simultaneously relieve persistent breathlessness and inflammation. HMDD can be considered as a potential non-steroidal anti-inflammatory drug in novel therapy design for the treatment of COPD and other inflammatory diseases.

Biomarker identification and pathway analysis of Astragalus membranaceus and Curcuma zedoaria couplet medicines on adenine-induced chronic kidney disease in rats based on metabolomics.

Background: Chronic kidney disease (CKD) is usually insidious, and most affected individuals are asymptomatic until the disease becomes advanced. The effective treatment of CKD would rely on the incorporation of multidisciplinary approaches. Astragalus membranaceus (AM) and Curcuma zedoaria (CZ) have been widely used in the treatment of CKD. However, the mechanism of AM and CZ in the treatment of CKD is still unclear. Methods: This study was designed to evaluate the effects of AM and CZ on adenine-induced rats and to investigate the underlying mechanism by using metabolomic analysis. Addition of 0.75% adenine to the diet of rats for 3 weeks induced the animal model of CKD. The rats in the treatment group were treated with AM and CZ (2.1 g/kg/day) for 4 weeks. Blood and kidney samples were collected for biochemical and histological examination. Ultra-high-performance liquid chromatography/Q Exactive HFX mass spectrometer (UHPLC-QE-MS) was applied to analyze metabolic profiling variations in the kidney. Results: The results showed that AM and CZ could significantly reduce serum creatinine (Scr) and blood urea nitrogen (BUN) levels in CKD rats and alleviate renal pathological injury. By comparing the endogenous components of the normal group and the model group in positive ion mode and negative ion mode, a total of 365 and 155 different metabolites were screened, respectively. A total of 117 and 73 metabolites with significantly different expressions were identified between model group and AM and CZ group in positive ion mode and negative ion mode, respectively. The pivotal pathways affected by AM and CZ included nicotinate and nicotinamide metabolism, and glycine, serine and threonine metabolism. Furthermore, significant changes in metabolites in CKD rats after AM and CZ therapies were observed, including L-Threonine, D-pantothenic acid, and nicotinamide. Moreover, we found that AM and CZ significantly reduced renal fibrosis and inflammation in CKD rats, which may be related to the regulation of SIRT1/JNK signaling pathway. Conclusion: In conclusion, AM and CZ significantly reduced renal fibrosis and inflammation in CKD rats, which may be related to the regulation of SIRT1/JNK signaling pathway. Furthermore, L-Threonine, D-pantothenic acid, and nicotinamide may be potential biomarkers for the progression and treatment of CKD.

Germacrone induces caspase-3/GSDME activation and enhances ROS production, causing HepG2 pyroptosis.

Liver cancer is a highly lethal malignancy. Despite considerable efforts made in recent years, the prognosis of patients with liver cancer remains poor. Curcuma zedoaria (known as Ezhu in Chinese) is widely prescribed in traditional Chinese medicine. Germacrone (GM) is a sesquiterpene constituent derived from the essential oil of Ezhu, and exerts anti-carcinogenic effects by inducing apoptosis in various cancer cells. The present study investigated the potential mechanism of GM in HepG2 cells. Cell Counting Kit-8, colony-formation and lactate dehydrogenase-release assays, as well as cell death assays using flow cytometry, were performed to evaluate HepG2 cell proliferation following GM treatment. HepG2 cells were transfected with caspase-3 small interfering RNA and then treated with GM. Caspase-3 expression levels were determined by reverse transcription-quantitative PCR and western blotting. The present study showed that GM inhibited the growth of HepG2 cells and induced the proteolytic cleavage of caspase 3, with concomitant cleavage of gasdermin E (GSDME), by markedly increasing the production of reactive oxygen species (ROS). This led to caspase 3-dependent cleavage of GSDME, thereby promoting pyroptosis in HepG2 cells. However, these changes were rescued by ROS scavengers, such as N-acetylcysteine. Furthermore, GM inhibited tumor growth by promoting the cleavage of caspase 3 and GSDME in HepG2 cell xenograft models. These results indicated that GM induced GSDME-dependent pyroptosis through caspase 3 activation, at least in part, by damaging the mitochondria and enhancing ROS production, thereby supporting the possible development of GM as a candidate for the prevention and treatment of liver cancer.

Curcuma zedoaria 50% methanol extracts increase adiponectin secretion by enhancing PPARγ mRNA expression in 3T3-L1 cells.

Curcuma zedoaria is a characteristic species of its genus that contains little to no curcuminoid. Here, we demonstrate that C. zedoaria extracts with 50% methanol increases adiponectin secretion into the media by enhancing PPARγ mRNA expression in 3T3-L1 cells. These results indicate that C. zedoaria may be useful for preventing/improving lifestyle-related diseases such as diabetes and atherosclerosis.

A new diphenylheptanoid from the rhizomes of Curcuma zedoaria.

A phytochemical investigation of the rhizomes of Curcuma zedoaria was carried out, leading to the isolation of a new diphenylheptanoid, zedoaroxane A (1), together with four known compounds (2-5). Their structures were elucidated based on NMR spectroscopic data. All isolated compounds possessed α-glucosidase inhibitory activity, with the IC50 values ranging from 35.2 to 89.0 µM, more potent than that of the positive control acarbose (IC50, 214.5 µM).

Validation of Unani concept of Abadal-i-Adwiya (drug substitution) by physicochemical standardization and hepatoprotective activity of Aristolochia rotunda Linn. and its substitute Curcuma Zedoaria Rosc. in albino Wistar rats.

OBJECTIVES: To validate the concept of abadal-i-adwiya (drug substitution) by evaluation of physicochemical standardization and hepatoprotective activity of Aristolochia rotunda & its substitute, Curcuma Zedoaria in albino Wistar rats. METHODS: Physicochemical standardization by estimation of moisture content, ash values and extractive values were carried out using standard methods. Hepatotoxicity was induced in albino Wistar rats using CCl4 1 mL/kg s. c. on alternate day for 14 days. Group I was served as Plain control and Group II as Negative control. Group III was administered silymarin 50 mg/kg p. o. while Group IV received HAE of A. rotunda 89.64 mg/kg p. o., and Group V was administered HAE of C. Zedoaria 45.73 mg/kg p. o. At the end of the study, serum bilirubin, AST (SGOT), ALT (SGPT) and ALP were estimated. The histopathology of liver was also carried out. RESULTS: The physicochemical parameters of both test drugs viz. moisture content, total ash, acid insoluble ash and water soluble ash were found within normal limit. The total serum bilirubin, direct bilirubin, AST (SGOT), ALT (SGPT) levels were significantly decreased in Test groups A and B when compared to the Negative and Standard controls. The microscopic examination of liver collected from animals of Group IV and Group V revealed significant recovery from hepatic toxicity compared to the Negative control. CONCLUSIONS: The study experimentation has revealed that C. Zedoaria may be used as a substitute for A. rotunda in the treatment of liver diseases. However, the outcome has to be further corroborated with phytochemical evaluation and clinical trials of both the drugs. Furthermore, the concept of drug substitute in Unani system of medicine is also validated in the light of above study.

Preliminary phytochemical analysis, antibacterial and anti-biofilm activities of Curcuma zedoaria (Christm.) Roscoe extracts

Aims@#Plant extracts are a rich source of natural compounds that have some degree of antimicrobial efficacy and have less side effects compared to antibiotics. The aim of this research was to screen the phytochemical compounds and investigate the potency of Curcuma zedoaria (Christm.) Roscoe rhizome (CZR) extracts to inhibit the growth and biofilm formation of some pathogenic bacteria.@*Methodology and results@#Antimicrobial and antibiofilm effects of CZR extracts in different solvents were examined by agar well diffusion and the broth microdilution method after phytochemical screening. The 95% ethanolic extract of CZR exhibited broad-spectrum antibacterial properties against Gram-negative and Gram-positive bacteria with inhibition zones of 7.25 ± 0.58-12.00 ± 0.26 mm and MIC values ranging from 50-200 mg/mL. The extract also showed rapid bacteriostatic and bactericidal activities towards Enterococcus faecalis DMST 4736 and Staphylococcus aureus ATCC 25923 by time-kill assays. Moreover, the 95% ethanolic extracts of CZR also acted as a potent anti-biofilm agent against E. faecalis DMST 4736, S. aureus ATCC 25923, S. epidermidis, Escherichia coli ATCC 25922, Klebsiella pneumoniae, Pseudomonas aeruginosa ATCC 27853 and Proteus mirabilis DMST 8212 (54.62 ± 0.30-71.25 ± 0.20% inhibition of biofilm formation). The bioactive potency of compounds of the crude 95% ethanolic extract (tannins, flavonoids, cardiac glycosides, steroids, terpenoids and alkaloids) play important roles in the observed antibacterial and anti-biofilm activities.@*Conclusion, significance and impact of study@#Curcuma zedoaria (Christm.) Roscoe extract had broad-spectrum antibacterial activity. The ethanolic CZR extract revealed bacteriostatic and bactericidal capacities, depending on time of exposure and concentration of the extracts. Thus, the present results indicate that C. zedoaria (Christm.) Roscoe rhizomes are a potential natural alternative antibacterial agent for preventing bacterial diseases.

Extract of Curcuma zedoaria R. prevents atherosclerosis in apolipoprotein E-deficient mice.

BACKGROUND/OBJECTIVES: Curcuma zedoaria R. (Zingiberaceae) has been used to treat headache, fever, and hypertension-related symptoms in Asian countries, including Korea, China, and Japan. We investigated whether dietary intake of a C. zedoaria extract (CzE) affected atherosclerosis in vivo. MATERIALS/METHODS: Apolipoprotein E-deficient (ApoE-/-) mice (n = 32) were fed a normal diet (ND), a high-cholesterol diet (HCD), an HCD containing CzE (100 mg/kg/day), or an HCD containing simvastatin (10 mg/kg/day) for 12 weeks. The anti-atherosclerotic effects were evaluated by observing changes in fatty streak lesions, immunohistochemical analysis, ex vivo fluorescence imaging, lipid profiles, and western blot analysis. RESULTS: The CzE-fed group showed a 41.6% reduction of atherosclerosis. Furthermore, CzE significantly reduced the levels of serum triglyceride, high-density lipoprotein, the chemokine (C-X3-C-motif) ligand 1, the adhesion molecules vascular cell adhesion molecule-1, intracellular adhesion molecule-1, and E-selectin; down-regulation of tumor necrosis factor-α, interleukin-6, high mobility group box-1, and cathepsin levels in the aortic sinuses and aortas of ApoE-/- mice were also observed. CONCLUSIONS: The results suggest that the inclusion of a water extract of C. zedoaria in a HCD is closely correlated with reducing the risk of vascular inflammatory diseases in an ApoE mouse model.

Effect of Curcuma zedoaria hydro-alcoholic extract on learning, memory deficits and oxidative damage of brain tissue following seizures induced by pentylenetetrazole in rat.

BACKGROUND: Previous studies have shown that seizures can cause cognitive disorders. On the other hand, the Curcuma zedoaria (CZ) has beneficial effects on the nervous system. However, there is little information on the possible effects of the CZ extract on seizures. The aim of this study was to investigate the possible effects of CZ extract on cognitive impairment and oxidative stress induced by epilepsy in rats. METHODS: Rats were randomly divided into different groups. In all rats (except the sham group), kindling was performed by intraperitoneal injection of pentylenetetrazol (PTZ) at a dose of 35 mg/kg every 48 h for 14 days. Positive group received 2 mg/kg diazepam + PTZ; treatment groups received 100, 200 or 400 mg/kg CZ extract + PTZ; and one group received 0.5 mg/kg flumazenil and CZ extract + PTZ. Shuttle box and Morris Water Maze tests were used to measure memory and learning. On the last day of treatments PTZ injection was at dose of 60 mg/kg, tonic seizure threshold and mortality rate were recorded in each group. After deep anesthesia, blood was drawn from the rats' hearts and the hippocampus of all rats was removed. RESULTS: Statistical analysis of the data showed that the CZ extract significantly increased the tonic seizure threshold and reduced the pentylenetetrazol-induced mortality and the extract dose of 400 mg/kg was selected as the most effective dose compared to the other doses. It was also found that flumazenil (a GABAA receptor antagonist) reduced the tonic seizure threshold compared to the effective dose of the extract. The results of shuttle box and Morris water maze behavioral tests showed that memory and learning decreased in the negative control group and the CZ extract treatment improved memory and learning in rats. The CZ extract also increased antioxidant capacity, decreased MDA and NO in the brain and serum of pre-treated groups in compared to the negative control group. CONCLUSION: It is concluded that the CZ extract has beneficial effects on learning and memory impairment in PTZ-induced epilepsy model, which has been associated with antioxidant effects in the brain or possibly exerts its effects through the GABAergic system.

Rapid plant regeneration in industrially important Curcuma zedoaria revealing genetic and biochemical fidelity of the regenerants.

The present investigation was carried out to establish an efficient and reproducible micropropagation protocol for the production of morphologically, genetically and chemically uniform plants of Curcuma zedoaria. Axillary bud explants of C. zedoaria were inoculated into MS basal medium supplemented with various combinations and concentrations of 6-benzyladenine (2.2-22.2 µM, BA), kinetin (2.3-23.2 µM, Kin), indole-3-acetic acid (2.9-11.4 µM, IAA), α-naphthalene acetic acid (2.7-10.2 µM, NAA) and adenine sulphate (33.9-203.6 µM, Ads). Almost 95% of rhizome buds sprouted on MS medium supplemented with 13.3 µM BA, 5.7 µM IAA and 63.9 µM Ads giving rise to an average of 12.89 ± 0.02 shoots within 6 weeks. However, the maximum number of roots (25.8 ± 0.07 roots per explant) was obtained on half strength MS medium supplemented with 7.4 µM of IBA after 4 weeks of inoculation. Morphological characteristics were similar in both conventionally propagated and micropropagated plants. Additionally, genetic homogeneity of in vitro plants was further confirmed through ISSR and flow cytometry analysis. A total of 27 ISSR primers were screened, out of which 13 ISSR primers generated 58 monomorphic and reproducible bands thereby confirming the genetic uniformity of obtained plants. The mean 2C DNA content of the mother plant (2.96 pg) was similar to that of in vitro derived plants (3.07 pg). Gas chromatography-mass spectrometry (GC-MS) analysis showed similarity in the qualitative profile of chemical constituents of essential oil and high-performance liquid chromatography analysis revealed no significant differences in curcumin content in the tissue culture regenerants and mother plants of C. zedoaria. Therefore, the present micropropagation protocol could be effectively employed to generate true to type plantlets of C. zedoaria.

Identification of a Quality Marker of Vinegar-Processed Curcuma Zedoaria on Oxidative Liver Injury.

Curcuma zedoaria (dry stenophora of Curcuma phaeocaulis Val., Curcuma kwangsiensis S. G. Lee et C. F. Liang, or Curcuma wenyujin Y. H. Chen et C.Ling) is a representative herb with clinical effects on liver diseases after being vinegar-processed. The crude Curcuma zedoaria and the processed Curcuma zedoaria (vinegar-boil) have been widely used as mixtures, but their equivalence has not been fully investigated. In this manuscript, quality markers of processed (vinegar-boil) Curcuma zedoaria were investigated by comparison of the compounds and hepatoprotective activities with the crude (three spices) ones. First, GC-MS-based untargeted metabolomics were applied to reveal the discriminatory components and discover potential markers. As a result, a total of six components were identified as potential markers. Then, the hepatoprotective activities were evaluated by dual cell damage models induced by a certain concentration of H2O2 or tertbutyl hydfroperoxide (t-BHP) (55 µM H2O2 or 40 µM t-BHP), which highlighted the potential of the processed Curcuma zedoaria on oxidative stress. Finally, epicurzerenone was identified as its quality marker on oxidative liver injury based on the above results and the cell-based biological assay. Overall, vinegar-processed Curcuma zedoaria was more suitable for the treatment of oxidative liver diseases, and epicurzerenone could be considered as its quality marker.

Ethnobotany, Phytochemistry and Traditional Uses of Curcuma spp. and Pharmacological Profile of Two Important Species (C. longa and C. zedoaria): A Review.

ETHNOPHARMACOLOGICAL RELEVANCE: The genus Curcuma, which is the most important source of curcumin, has been widely used in different traditional medicines. Various species of Curcuma have long been used for several purposes such as healing wounds, liver disorders, jaundice and also as a blood purifier. AIM OF THE STUDY: This review focused on the ethnopharmacological uses and phytochemical aspects of Curcuma. Additionally, in this study, the different properties of two species of Curcuma in Islamic Traditional Medicine (ITM), C. longa and C. zedoaria, as well as their pharmacological aspects in modern medicine are reviewed. MATERIALS AND METHODS: ITM literatures were searched to find Curcuma's applications. Also, electronic databases including PubMed and Scopus were searched to obtain studies giving any in vitro, in vivo or human evidence of the efficacy of C. longa and C. zedoaria in the treatment of different diseases. ChemOffice software was used to find chemical structures. RESULTS: The analysis showed that ethno-medical uses of Curcuma have been recorded for centuries. Approximately, 427 chemical compounds have been isolated and identified from Curcuma spp. This genus is rich in flavonoids, tannins, anthocyanin, phenolic compounds, oil, organic acids and inorganic compounds. Curcumin is one of the main active ingredients in Curcuma which has strong anti-inflammatory and antioxidant effects. Besides, pharmacological studies have indicated wide range of Curcuma's activities, such as hepato-protective, antifungal, antihypertensive and neuroprotective. CONCLUSIONS: In this study, we reviewed various studies conducted on ethno-medicinal, ITM properties and photochemistry of Curcuma spp. Also, pharmacological activities of two species, C. longa and C. zedoaria are summarized. Pre-clinical investigations have demonstrated some of the traditional aspects of Curcuma, such as wound healing, anti-arthritic, anti-tumor and liver protective activities. These could be related to antioxidant and anti-inflammatory properties of Curcuma which might be due to high amounts of phenolic compounds. Curcuma is mentioned to have neural tonic properties in ITM which have been confirmed by some animal studies. Considering various preclinical studies on C. longa and C. zedoaria and their active ingredient, curcumin, randomized controlled trials are warranted to confirm their promise as a clinically effective hepato and neuro-protective agents.

Sesquiterpenes from Curcuma zedoaria rhizomes and their cytotoxicity against human gastric cancer AGS cells.

Curcuma zedoaria rhizome (Zingiberaceae) is a well-known traditional medicinal plant used in Ayurvedic and traditional Chinese medicine to treat various cancers. This study aimed to identify the cytotoxic components from C. zedoaria rhizomes that act against gastric cancer, which is the third leading cause of death from cancer worldwide because the MeOH extract of C. zedoaria rhizome was found to show a cytotoxic effect against gastric cancer AGS cells. Repeated column chromatography and semi-preparative HPLC purification were used to separate the components from the C. zedoaria MeOH extract. Two new sesquiterpenes, curcumenol-9,10-epoxide (1) and curcuzedoalide B (2), and 12 known related sesquiterpenes (3-14) were isolated from the C. zedoaria MeOH extract. The structures of new compounds were determined by 1D and 2D NMR spectroscopic experiments and HR-ESIMS, and quantum chemical ECD calculations. The cytotoxic effects of the isolated compounds were measured in human gastric cancer AGS cells using an MTT cell viability assay. Compounds 9, 10, and 12 exhibited cytotoxic effects against gastric cancer AGS cells, with IC50 values in the range of 212-392 µM. These findings provide further experimental scientific evidence to support the traditional use of C. zedoaria rhizomes for the treatment of cancer. Curcumenol (9), 4,8-dioxo-6ß-methoxy-7α,11-epoxycarabrane (10), and zedoarofuran (12) were identified as the main cytotoxic components in C. zedoaria rhizomes.

Evaluation of the Anti-Trypanosomal Activity of Vietnamese Essential Oils, with Emphasis on Curcuma longa L. and Its Components.

Human African trypanosomiasis (HAT), known as sleeping sickness and caused by Trypanosoma brucei, is threatening low-income populations in sub-Saharan African countries with 61 million people at risk of infection. In order to discover new natural products against HAT, thirty-seven Vietnamese essential oils (EOs) were screened for their activity in vitro on Trypanosoma brucei brucei (Tbb) and cytotoxicity on mammalian cells (WI38, J774). Based on the selectivity indices (SIs), the more active and selective EOs were analyzed by gas chromatography. The anti-trypanosomal activity and cytotoxicity of some major compounds (isolated or commercial) were also determined. Our results showed for the first time the selective anti-trypanosomal effect of four EOs, extracted from three Zingiberaceae species (Curcuma longa, Curcuma zedoaria, and Zingiber officinale) and one Lauraceae species (Litsea cubeba) with IC50 values of 3.17 ± 0.72, 2.51 ± 1.08, 3.10 ± 0.08, and 2.67 ± 1.12 nL/mL respectively and SI > 10. Identified compounds accounted for more than 85% for each of them. Among the five major components of Curcuma longa EO, curlone is the most promising anti-trypanosomal candidate with an IC50 of 1.38 ± 0.45 µg/mL and SIs of 31.7 and 18.2 compared to WI38 and J774 respectively.

Variations in the Volatile Compositions of Curcuma Species.

Curcuma species have been cultivated in tropical and subtropical regions in Asia, Australia, and South America for culinary as well as medicinal applications. The biological activities of Curcuma have been attributed to the non-volatile curcuminoids as well as to volatile terpenoids. Curcuma essential oils have demonstrated a wide variety of pharmacological properties. The objective of this work was to examine the variation in the compositions of Curcuma rhizome essential oils. In this work, the volatile oils from C. longa and C. zedoaria were obtained and analyzed by gas chromatography-mass spectrometry. The chemical compositions of C. longa and C. zedoaria essential oils, including those reported in the literature, were analyzed by hierarchical cluster analysis. In addition, cluster analyses of the chemical compositions of C. aromatica and C. aeruginosa from the literature were also carried out. Curcuma longa volatiles were dominated by α-turmerone, curlone, ar-turmerone, ß-sesquiphellandrene, α-zingiberene, germacrone, terpinolene, ar-curcumene, and α-phellandrene and showed four distinct chemical clusters. C. zedoaria rhizome oil contained 1,8-cineole, curzerenone/epi-curzerenone, α-copaene, camphor, ß-caryophyllene, elemol, germacrone, curzerene, and ß-elemene and showed two different chemical types. C. aromatica had three clearly defined clusters, and C. aeruginosa had three types.

Bioactivity-based analysis and chemical characterization of anti-inflammatory compounds from Curcuma zedoaria rhizomes using LPS-stimulated RAW264.7 cells.

Inflammation is not only a self-defense response of the innate immune system, but also the pathogenesis mechanism of multiple diseases such as arthritis, neurodegeneration, and cancer. Curcuma zedoaria Roscoe (Zingiberaceae), an indigenous plant of India, has been used traditionally in Ayurveda and folk medicine. As part of our ongoing efforts to screen traditional medicinal plants exhibiting pharmacological potential and to characterize the compounds involved, we examined the anti-inflammatory effects of the MeOH extract of C. zedoaria rhizomes using lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage cells and found that MeOH extract inhibited the synthesis of nitric oxide (NO) in a dose-dependent manner (IC50: 23.44 ±â€¯0.77 µg/mL). In our efforts to characterize the compounds responsible for these anti-inflammatory effects, bioactivity-guided fractionation of the MeOH extract and chemical investigation of its active hexane-soluble fraction led to the successful isolation of five sesquiterpenes (1-5), the structures of which were elucidated by NMR spectroscopic analysis and LC/MS analysis. Among them, curcuzedoalide (5) exhibited potent inhibitory effects on NO synthesis (IC50: 12.21 ±â€¯1.67 µM) and also suppressed pre-inflammatory protein expression of iNOS and COX-2. Curcuzedoalide (5) was thus determined to be a contributor to the anti-inflammatory effect of C. zedoaria rhizomes and could be a potential candidate for therapeutic applications.

Identification and Characterization of Genes in the Curcuminoid Pathway of Curcuma zedoaria Roscoe.

BACKGROUND: Curcuminoid genes have an important role in the biosynthesis of curcumin, a valuable bioactive compound, in Curcuma species. However, there have not been any reports of these genes in Curcuma zedoaria. OBJECTIVE: The present work reports on the isolation of genes encoding enzymes in curcuminoid metabolic pathway and their expression in C. zedoaria. METHOD: The primers were designed from untranslation regions of DCS, CURS1, CURS2 and CURS3 genes which are involved in curcuminoid biosynthesis in C. longa to isolate the corresponding fulllength genes in C. zedoaria. RT-PCR amplification and HPLC analysis are used to estimate the expression of genes and biosynthesis of curcumin in both rhizome and callus. RESULTS: The results showed that all four genes from C. zedoaria (named CzDCS, CzCURS1, CzCURS2 and CzCURS3) and C. longa have a high identity (approximately 99%) and lengths of genes from C. zedoaria are 1382, 1240, 1288 and 1265 nu, respectively. CzCURS1, 2 and 3 genes have one intron while CzDCS has two introns. RT-PCR amplification indicated that curcuminoid genes expressed mRNA in rhizome and callus of C. zedoaria. Curcumin, a major component of curcuminoids, was also found in callus by HPLC analysis. CONCLUSION: The sequence information of DCS and CURS1-3 genes in C. zedoaria will be very valuable for a subsequent study on the effects of elicitors on the transcription of genes involved in curcuminoid biosynthesis pathway.

Discovery of ß2- adrenoceptor agonists in Curcuma zedoaria Rosc using label-free cell phenotypic assay combined with two-dimensional liquid chromatography.

Traditional Chinese Medicines (TCMs) have been widely used in clinical practice, and provided a rich source for discovering new drug leads. However, efficient identification of active molecules responsible for the therapeutic effects of complex TCMs is still highly challenging. Here, we combined label-free cell phenotypic assay with two dimensional liquid chromatography (2DLC) to identify potential ß2-adrenoceptor (ß2-AR) agonists related to anti-asthmatic effect of Curcuma zedoaria Rosc (C.zedoaria), a commonly used TCM. The ethyl acetate extract of C.zedoaria was first fractionated into 26 fractions. Label-free cell phenotypic profiling was then used to locate the active sites. Orthogonal second-dimensional (D2) separation was performed on two fractions displaying agonistic effect at the ß2-AR, combined with screening of the D2 fractions to track the activity. Finally, this approach led to the isolation of three known diarylheptanoids, among which diarylheptanoid b exhibited the most potent agonistic activity with an EC50 value of 5.93 µM. This result was further demonstrated through the chemical synthesis of diarylheptanoid b. It is the first time to discover that diarylheptanoids could activate the ß2-AR, which may be responsible for the anti-asthmatic effect of C.zedoaria observed traditionally and in clinical application. This study also demonstrates the potential of this integrated strategy for identifying active ingredients and determining the basis of therapeutic materials in complex TCMs.

Chemical Composition and Biological Activities of Essential Oils of Curcuma Species.

Members of the genus Curcuma L. have been used in traditional medicine for centuries for treating gastrointestinal disorders, pain, inflammatory conditions, wounds, and for cancer prevention and antiaging, among others. Many of the biological activities of Curcuma species can be attributed to nonvolatile curcuminoids, but these plants also produce volatile chemicals. Essential oils, in general, have shown numerous beneficial effects for health maintenance and treatment of diseases. Essential oils from Curcuma spp., particularly C. longa, have demonstrated various health-related biological activities and several essential oil companies have recently marketed Curcuma oils. This review summarizes the volatile components of various Curcuma species, the biological activities of Curcuma essential oils, and potential safety concerns of Curcuma essential oils and their components.

Wound healing effects of a Curcuma zedoaria polysaccharide with platelet-rich plasma exosomes assembled on chitosan/silk hydrogel sponge in a diabetic rat model.

One homogeneous polysaccharide (ZWP) was successfully isolated and purified from the rhizomes of Curcuma zedoaria and the aim of present study was to determine the potential of chitosan/silk hydrogel sponge loaded with platelet-rich plasma exosomes (PRP-Exos), ZWP or PRP-Exos/ZWP on wound-healing in diabetic rats. An in vivo diabetic wound healing study showed that separate or combined treatments all resulted in a wound contraction in diabetic rats, as evidenced by a decrease of ulcer and an increase of epidermal thickness, but PRP-Exos/ZWP combination therapy was more successful in wound closing than either PRP-Exos or ZWP single administration. This could be due to the upregulation of collagen synthesis and deposition, as well as angiogenesis at the wound site. In addition to this, no side effects was observed in all treated groups during healing process. These findings provide a mechanistic basis for PRP-Exos/ZWP as a potential therapeutic strategy to accelerate skin repair in diabetes.