An Investigation into the Phytochemical Content and Antioxidant, Antidiabetic, and Wound-Healing Activities of Curculigo latifolia Found in Brunei Darussalam.

This present study aimed to investigate the phytochemical content and antioxidant and antidiabetic activities of Curculigo latifolia leaves (CL) and C. latifolia roots (CR) found in Brunei Darussalam. Phytochemical screening showed that CL and CR extracts contain saponins, tannins, glycosides, and terpenoids. CR showed higher total phenolic content (TPC), but lower total flavonoid content (TFC) when compared to CL. The high TPC in CR contributed to its potent radical scavenging activity (RSA) against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and strong ferric reducing antioxidant power (FRAP). Additionally, CR exerted significant inhibition of ∝-glucosidase and ∝-amylase, suggesting a potential link between the chemical compounds and its antioxidant and antidiabetic effects. In the animal study of antihyperglycemic activity, treatment with 250 mg/kg body weight (b.w.) of the CL extract normalised the blood glucose levels and improved body weight gain of alloxan-induced diabetic rats within 14 weeks. Furthermore, our investigation into the wound-healing effects of young C. latifolia leaves (YCL) and matured C. latifolia leaves (MCL) showed a significant reduction in wound size on Day 3, 5, and 7 of the experimental study, indicating its wound-healing potential. Based on our findings, C. latifolia can be consumed as part of a balanced diet due to its antioxidant and antidiabetic properties.

Effect of light and cytokinin on growth and curculin gene expression of Curculigo latifolia on in vitro culture.

Despite being valuable for producing a natural sweetener Curculin, Curculigo latifolia has a low growth and difficult to domestificate. So, to solve this problem, propagation on in vitro culture will be an alternative method to propagated this spesies under different cytokinins and light condition. Cytokinins and light has major role in organogenesis, growth and gene expression of many species. Thus, in this study, we aimed to improve the Curculigo latifolia growth on in vitro condition and expression of curculin gene by combining cytokinins addition and different light exposure. Four weeks seedlings were sub-cultured into medium (MS free hormone) containing 3 mg/L benzyladenine (BA) and various concentrations of meta-Topolin (mT) including 0.1 mg/L, 0.5 mg/L, and 5 mg/L. The cultures then incubated under different light types (red, blue, white LED lights and white fluorescence light) with 16-h light/ 18-h dark photoperiod for 14 weeks at 25 ± 2°C. Several parameters, including plant height, leaf number, chlorophyll contents, stomatal structure, and density and curculin expression, were observed every week. Unexpectedly, our results showed that C. latifolia growth displayed significant improvement when it was treated under white LED light without any additional cytokinins. In sum, white LED light further improves plantlets phenotype, such as plant height, leaf number, chlorophyll production, and stomatal number and structure, whereas, red LED light lead to a decreased phenotypes but increase the curculin gene expression.

Network pharmacology and experimental validation methods to reveal the active compounds and hub targets of Curculigo orchioides Gaertn in rheumatoid arthritis.

BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disease that can lead to joint destruction and deformity. Curculigo orchioides Gaertn (CO) was previously revealed to play a significant role in RA treatment. However, the main active ingredients and molecular mechanisms of CO in regulating RA are still unclear. METHODS: The active ingredients of CO were obtained from the Traditional Chinese Medicine Systems Pharmacology database and published literature. The targets corresponding to these compounds and the targets linked to RA were collected from public databases. The "ingredient-target" and "protein-protein interaction" networks were constructed to screen the main active ingredients and hub targets of CO in the treatment of RA. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment assays were used to elucidate the potential pharmacological mechanism of CO in RA. Molecular docking was performed to detect the binding between the main active ingredients and hub targets. Collagen-induced arthritis rats were used to validate the hub targets of CO against RA. RESULTS: Network pharmacological topology analysis showed that caffeine, 2,4-dichloro-5-methoxy-3-methylphenol, curculigoside, orcinol glucoside, and orcin were the main active ingredients of CO, and matrix metalloproteinase 9 (MMP9), transcription factor AP-1 (JUN), prostaglandin-endoperoxide synthase 2 (PTGS2), brain-derived neurotrophic factor, and receptor-type tyrosine-protein phosphatase C were the hub targets of CO for RA treatment. Molecular docking revealed that curculigoside and orcinol glucoside had effective binding potential with MMP9, JUN, and PTGS2, respectively. In vivo experiments demonstrated that CO alleviated RA symptoms and inhibited the expression of MMP9, JUN, and PTGS2 proteins. CONCLUSIONS: Our study demonstrates the main active ingredients and potential targets of CO against RA, laying an experimental foundation for the development and application of CO as an anti-RA drug.

Pharmacognostical and Phytochemical Studies and Biological Activity of Curculigo latifolia Plant Organs for Natural Skin-Whitening Compound Candidate.

Curculigo latifolia (family Amaryllidaceae) is used empirically for medicinal purposes. It is distributed throughout Asian countries, especially Indonesia. This study aimed at standardizing the C. latifolia plant, analyzing its phytochemical profile, and evaluating its pharmacological effects. The powder from each organ (root, stem, and leaves) was standardized organoleptically and microscopically. Samples were extracted by graded maceration using hexane, ethyl acetate, and ethanol. The extracts were determined for total phenolic content (TPC) and total flavonoid content (TFC). Antioxidant (radical scavenging and metal ion reduction) and antityrosinase activities were determined by spectrophotometric methods. Extracts were analysed for phytochemical profiles by LC-ESI-MS. The highest TPC and TFC were found in the ethanolic extract of the root organ (68.63 ± 2.97 mg GAE/g) and the ethyl acetate extract of the stem (14.33 ± 0.71 mg QE/g extract). High antioxidant activities were found in the ethanolic root extract (20.42 ± 0.33 µg/mL) and ethanolic stem extract (45.65 ± 0.77 µg/mL) by DPPH• and NO• assays, respectively. The ion reduction activity (by CUPRAC assay) was most significant in the ethyl acetate stem extract (390.42 ± 14.49 µmol GAEAC/g extract). Ethanolic root extract was the most active in inhibiting tyrosinase (IC50 value of 108.5 µg/mL). The correlation matrix between TPC and antioxidant activities showed a moderate to robust correlation, whereas the TPC and antityrosinase activity showed a robust correlation. The TFC and antioxidant or antityrosinase activities showed a weak to moderate correlation. The LC-ESI-MS data identified major phenols in the active extracts, including methyl 3-hydroxy-4-methoxy-benzoate, quercetin, 4-O-caffeoylquinic acid-1, and curculigoside. Overall, this study suggests that extracts from the C. latifolia plant offer potent antioxidant and antityrosinase activities, allowing them to be used as natural antioxidants and candidates for skin-lightening compounds.

Molecular networking-assisted isolation of chlorophenolic glycosides from the rhizomes of Curculigo orchioides and their inhibitory effect on α-glucosidase.

Molecular networking analysis and in silico tools, such as Network Annotation Propagation (NAP) and MolNetEnhancer, were applied to explore bioactive constituents present in the ethyl acetate-soluble fraction of the rhizomes of Curculigo orchioides. Among the molecular networks, the most abundant cluster was classified as a phenolic glycoside using the ClassyFire module of MolNetEnhancer. Further, the major node in this cluster was accurately predicted as curculigine A using the in silico fragment analysis tool, NAP. Six undescribed chlorophenolic glycosides (1-6) and 11 known phenolic glycosides were isolated, using molecular networking-assisted isolation methods, and their structures were elucidated using 1D, 2D-NMR and HRESIMS. In particular, the structures of the isolated chlorophenolic glycosides, which have non-protonated aromatic rings, were determined using various NMR experiments, such as 1D-selective NOE, ROESY, and LR-HMBC, and acid hydrolysis. All isolated compounds were examined to determine their inhibitory effects on α-glucosidase and compounds 3, 8, 10, 11, 13, 14, and 16 revealed the IC50 values ranging from 19.6 to 35.5 µM. Their structure-activity relationships were also evaluated based on the analysis of their inhibitory effects and performance of molecular docking simulations.

Curculigo orchioides polysaccharide COP70-1 stimulates osteogenic differentiation of MC3T3-E1 cells by activating the BMP and Wnt signaling pathways.

The crude polysaccharide CO70 isolated from Curculigo orchioides could alleviate ovariectomy-induced osteoporosis in rats. To clarify the bioactive components, a new heteropolysaccharide (COP70-1) was purified from CO70 in this study, which was consisted of ß-D-Manp-(1→, →4)-α-D-Glcp-(1→, →4)-ß-D-Manp-(1→, →3,4)-ß-D-Manp-(1→, →4,6)-ß-D-Manp-(1→, and →4,6)-α-D-Galp-(1→. COP70-1 significantly promoted the osteoblastic differentiation of MC3T3-E1 cells through improving alkaline phosphatase activity, the deposition of calcium as well as up-regulating the expression of osteogenic markers (RUNX2, OSX, BSP, OCN, and OPN). Furthermore, COP70-1 stimulated the expression of critical transcription factors of the BMP and Wnt pathways, including BMP2, p-SMAD1, active-ß-catenin, p-GSK-3ß, and LEF-1. In addition, LDN (BMP pathway inhibitor) and DKK-1 (Wnt pathway inhibitor) suppressed the COP70-1-induced osteogenic differentiation of MC3T3-E1 cells. Therefore, COP70-1 was one of the bioactive constituents of C. orchioides for targeting osteoblasts to treat osteoporosis by triggering BMP/Smad and Wnt/ß-catenin pathways.

Decipher the pharmacological mechanisms of raw and wine-processed Curculigo orchioides Gaertn. on bone destruction in rheumatoid arthritis rats using metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Curculigo orchioides Gaertn. (CO), a traditional Chinese herb recorded in Chinese Pharmacopoeia, can nourish kidney yang, strengthen bones, and dispell cold-dampness. Raw CO (rCO) and wine-processed CO (pCO), the main processed products of CO for clinical application, show differences in nourishing kidney yang and ameliorate osteoporosis. However, the difference in efficacy and mechanism of rCO and pCO on bone destruction in rheumatoid arthritis (RA) remain unclear. AIM OF THE STUDY: To compare the pharmacodynamics of rCO and pCO in the treatment of bone destruction in RA and to reveal the potential mechanism by which rCO and pCO exert effects by metabolomics approach. MATERIALS AND METHODS: Ultra-high performance liquid chromatography Q exactive mass spectrometry (UHPLC-Q-Exactive-MS) combined with multivariate data analysis was applied to identify the differential chemical components in rCO and pCO. Collagen-induced arthritis (CIA) rats were orally administrated with different doses of rCO and pCO for 4 weeks. The body weight, paw swelling, arthritis scores, serum inflammatory cytokines concentration, knee tumor necrosis factor (TNF)-α, interleukin (IL)-6 protein levels, and inflammatory cell infiltration were determined to investigate the effects of rCO and pCO on arthritic symptoms and inflammatory responses in CIA rats. The effects of rCO and pCO on bone destruction were assessed using safranin O-fast green and tartrate-resistant acid phosphatase (TRAP) staining, immunohistochemical analysis of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) proteins, and micro-computed tomography (micro-CT) in rats. In addition, metabolomics was performed to explore the mechanism of rCO and pCO against bone destruction in RA. RESULTS: A total of 41 chemical constituents were identified in both rCO and pCO, 9 of which were screened out as discriminatory compounds. According to the pharmacodynamic assays, pCO exhibited a stronger effect than rCO in attenuating the severity of arthritis, reducing inflammation, and inhibiting bone destruction. The metabolomics results showed that pentose phosphate pathway was the key metabolic pathways regulated by rCO, while pCO regulated multiple metabolic pathways including phenylalanine metabolism pathways, phenylalanine, tyrosine and tryptophan biosynthesis, taurine and hypotaurine metabolism, and glycerophospholipid metabolism pathways. CONCLUSION: pCO displayed a better effect on alleviating bone destruction in RA was than rCO. This might be associated with that pCO can decrease inflammation in RA through regulating more metabolism pathways.

In vitro somatic embryo productions from Curculigo orchioides derived gold nanoparticles: Synthesis, characterization, its biomedical applications, and their eco-friendly approaches to degradation of methylene blue under solar light irradiations.

Curculigo orchioides rhizome explants were employed to develop a rapid and effective strategy for increased plant regeneration using somatic embryogenesis. Direct somatic embryo development was shown on rhizome explants cultivated on Murashige and Skoog (MS) making with 2, 4-D (1.0-3.0 mg/L). Rhizome explants cultivated on MS media supplemented with 2.0 mg/L 2, 4-D yielded the highest frequency of embryogenesis (87.5%) and the maximum number of somatic embryos (1596.7/explant). Somatic embryo germination was accomplished using MS media with 2.0 mg/L 6-benzylaminopurine (BAP). With an 80% survival rate, the germination plantlets were acclimated in the greenhouse. The current study is the first evidence of the efficacy of in vitro-produced plants and C. orchioides somatic embryo callus cultures of stable gold nanoparticles. The UV-Vis spectrophotometric absorbance, at 510 nm, revealed the absorption spectra of the AuNPs. The FT-IR revealed functional groups and reaction processes in green AuNP formation. High-resolution transmission electron microscopy (HR-TEM) was used to assess the surface morphology and structure of the AuNPs after their elemental composition was determined using a dispersive energy X-ray (EDAX) spectrum. The average size of AuNPs was around 35 nm in diameter. The crystalline nature of the AuNPs was investigated by X-ray diffraction (XRD). The highest growth inhibition was found for C. orcthioides against Klebsiella pneumoniae (17.5 mm) and Serratia marcescens (16.5 mm). The AuNPs exhibited antioxidant activity against free radicals such as DPPH and ABTS. Furthermore, the cytotoxicity of AuNPs was assessed, and inhibitory concentration (IC50) was 20 µg/mL and 80 µg/mL for breast carcinoma (MDA-MB-231) and Vero cell lines. The degradation of methylene blue measures the photocatalytic activity of the manufactured AuNPs when subjected to visible sunlight (MB). Thus, the result showed a maximum degradation efficiency of MB (84%).

Toxicity Assessment of Curculigo orchioides Leaf Extract Using Drosophila melanogaster: A Preliminary Study.

Curculigo orchioides is used in Indian and Chinese traditional medicinal systems for various health benefits. However, its toxicological effects are mostly unknown. This study assesses the potential toxicity of aqueous leaf (A.L.) extract of C. orchioides using Drosophila melanogaster as an experimental model. Preliminary phytochemical tests were followed by the Fourier transform infrared (FTIR) tests to identify the functional group in the A.L. extract of C. orchioides. Drosophila larvae/adults were exposed to varying concentrations of C. orchioides A.L. extract through diet, and developmental, lifespan, reproduction, and locomotory behaviour assays were carried out to assess the C. orchioides toxicity at organismal levels. The cellular toxicity of A.L. extract was examined by analysing the expression of heat shock protein (hsps), reactive oxygen species (ROS) levels, and cell death. The FTIR analysis showed the presence of functional groups indicating the presence of secondary metabolites like saponins, phenolics, and alkaloids. Exposure to A.L. extract during development resulted in reduced emergence and wing malformations in the emerged fly. Furthermore, a significant reduction in reproductive performance and the organism's lifespan was observed when adult flies were exposed to A.L. extract. This study indicates the adverse effect of C. orchioides A.L. extract on Drosophila and raises concerns about the practice of indiscriminate therapeutic use of plant extracts.

Discrimination of Curculigo orchioides Rhizoma and Curculigo glabrescens Rhizoma using stable isotope and mineral element analyses coupled with chemometrics.

Correct species identification is crucial for ensuring the quality, safety, and efficacy of herbal medicine. Market research indicates that Curculigo glabrescens Rhizoma (CGR) was the major counterfeit of the medicine Curculigo orchioides Rhizoma (COR). To accurately discriminate COR and CGR remains a challenge, and it becomes even more difficult when the herbs have been heavily processed into a powder. In this work, combined with high performance liquid chromatography analysis, a novel component in CGR was discovered, and two stable isotopes (N%, C%, δ15N, δ13C) and nineteen mineral elements were determined along with multivariate statistical analysis to distinguish the authentic COR samples and counterfeit CGR samples. The results showed that there were significant differences between the mean value of N%, δ15N and δ13C according to the botanical origins. In addition, these two species can be differentiated by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) analysis. A linear discriminant analysis (LDA) model with a good classification rate (100%) and cross-validation rate (100%) was established. Hence, stable isotope and mineral element contents combined with chemometrics analysis could be considered as an effective and reliable method for discriminating the source species of COR and CGR.

Structural characterization and anti-osteoporosis activity of two polysaccharides extracted from the rhizome of Curculigo orchioides.

Curculigo orchioides is widely used to treat osteoporosis in China. In this study, we identified the active substances in the crude polysaccharide (CO50) from C. orchioides that had anti-osteoporosis activity in vivo. Two polysaccharides, COP50-1 and COP50-4, were purified from CO50. Based on structural analysis, COP50-1 was composed of α-D-Glcp-(1→, ß-D-Galp-(1→, →4)-α-D-Glcp-(1→, →3,4)-α-D-Glcp-(1→, →4,6)-α-D-Glcp-(1→, →4,6)-ß-D-Manp-(1→, whereas COP50-4 was composed of α-L-Araf-(1→, →2)-α-L-Rhap-(1→, ß-D-Manp-(1→, α-D-Galp-(1→, →2,4)-α-L-Rhap-(1→, →2)-ß-D-Manp-(1→, →4)-α-D-GlcAp-(1→, →3)-α-D-GalAp-(1→, →4,6)-α-D-Galp-(1→, →2,3,6)-ß-D-Manp-(1→, →2,3,5)-α-L-Araf-(1→, →2,5)-α-L-Araf-(1→, →4)-α-D-Glcp-(1→ and →3)-α-D-Galp-(1→. Pharmacological assessment revealed that COP50-1 had no obvious osteogenic activity. However, COP50-4 (0.5 µM) significantly enhanced the differentiation and mineralization of osteoblasts in vitro. Moreover, the effect of COP50-4 was greater than that of 17ß-estradiol. Therefore, COP50-4 may be an effective component of CO50 that has great potential for development as an alternative drug for the treatment of osteoporosis.

Norlignans and phenolics from genus Curculigo protect corticosterone-injured neuroblastoma cells SH-SY5Y by inhibiting endoplasmic reticulum stress-mitochondria pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The plants of genus Curculigo are divided into the Section Curculigo and the Section Capitulata, which are mainly distributed in southeastern and southwestern China. Various ancient chinese books record that these plants were used as an important herb for tonifying kidney yang. Traditional Chinese medicine often draws on this property to treat depression syndrome. Thus genus Curculigo has potential for the treatment of neurodegenerative diseases (ND). The study showed that phenolics were the main characteristic components of plants in the Section Curculigo, represented by orcinol glucoside and curculigoside; the norlignans, with Ph-C5-Ph as the basic backbone, were the main characteristic components of the Section Capitulata. However, there is a lack of sufficient scientific evidence as to whether these two types of ingredients have neuroprotective effects. AIM OF THE STUDY: To determine the neuroprotective effects of phenolics and norlignans in genus Curculigo on human neuroblastoma cells SH-SY5Y. To discuss their structure-activity relationship and screen for compounds with high activity and neuroprotective effects. To reveal that the amelioration of endoplasmic reticulum (ER) stress by two classes of compounds is mediated by the PERK/eIF2α/ATF4 pathway. MATERIALS AND METHODS: The cytotoxicity of 17 compounds was assayed by MTT. SH-SY5Y cells were damaged by corticosterone (Cort) (200 µM) for 24 h and then co-administered with 17 compounds (0.1-100 µM) and Cort (200 µM) for 24 h. Cell survival was determined by MTT assay. Apoptosis rate, mitochondrial membrane potential (MMP) and intracellular reactive oxygen species (ROS) levels were detected using flow cytometry. Intracellular Ca2+ levels were detected using a fluorescent probe. Cellular mitochondrial and ER damage was observed using transmission electron microscopy (TEM). ER stress and apoptotic pathway-related proteins (BiP, CHOP, cleaved caspase-3, cleaved caspase-9, Bax/Bcl-2), and the expression level of PERK/eIF2α/ATF4 pathway was measured via western blot (WB). RESULTS: The experimental data showed that Cort treatment of SH-SY5Y cells resulted in decreased cell survival and increased apoptosis, mitochondrial depolarization, ROS, and intracellular Ca2+ levels. The co-action of 17 compounds and Cort for a period of time significantly increased cell survival. Compounds 3, 7, 12, 13 also reduced apoptosis rate, mitochondrial depolarization, ROS and intracellular Ca2+ levels in the subsequent experiments. In addition, TEM observed that Cort caused mitochondrial and ER damage, and the damage was improved after treatment. WB analysis obtained that Cort increased the expression of apoptotic and ER stress-related proteins and activated pathway expression. However, in the presence of compounds 3, 7, 12, 13, the expression of BiP, CHOP, cleaved caspase-3, cleaved caspase-9, and Bax/Bcl-2 was significantly reduced, and the phosphorylation of PERK and eIF2α and the expression of ATF4 were inhibited. CONCLUSION: This study found that one phenolic (3) and three norlignans (7, 12, 13) from genus Curculigo have significant neuroprotective effects. The results of the structure-activity relationship indicated that the glucosyl polymeric norlignans and the phenolics with benzoic acid as the parent nucleus were more active. The neuroprotective effect of three norlignans is the latest discovery. This finding has important research value in the field of prevention and treatment of neurodegenerative diseases.

GC-MS analysis of curculigo orchiodes and medicinal herbs with cytotoxic, hepatoprotective attributes of ethanolic extract from Indian origin.

OBJECTIVES: Liver illnesses are a major public health issue all over the world. Medicinal plants constituents a viable alternative for the development of phytopharmaceuticals with hepatoprotective activity in order to solve some of these health-related problems. The present study is focused on the phytochemical and biological investigation on Indian traditional medicinal plant extracts, for their cytotoxic and hepatoprotective activity. The isolated compounds showed the presence of phenolic constituents which lead to cytotoxicity and hepatoprotective activity of medicinal plant. Cancer causes about 13% of all human deaths in 2007 (7.6 million) (American Cancer Society and WHO December 2006-07). The American Cancer Society estimates that 12,990 new cases of cervical cancer will be diagnosed in the United States year 2016. Cancer-related deaths are expected to increase, with an estimated 11.4 million deaths in 2030. METHODS: The ethanolic extracts of Centella asiatica, Myristica fragrans, Trichosanthes palmata, Woodfordia fruticosa, Curculigo orchioides were evaluated against HEP-G2 cell lines for hepatoprotective activity and Curculigo orchioides was further promoted for the isolation of secondary metabolites based on inhibitory concentration. RESULTS: The ethanolic extracts of C. asiatica, M. fragrans, T. palmata, W. fruticosa, Curculigo orchioides shown significant cytotoxic activity (IC50≤100 µg/mL). The plant extracts also shown significant hepatoprotective activity in a dose dependent manner when tested against HEP-G2 cell lines and cytotoxicity studies against HeLa and HEP-G2 cells. CONCLUSIONS: The extract of Curculigo orchiodes rhizome showed significant cytotoxicity results. Hence the Curculigo orchiodes rhizome was selected for further phytochemical studies to isolate active compounds and their Characterization by GCMS.

Phytochemistry and Pharmacological Activity of Plants of Genus Curculigo: An Updated Review Since 2013.

The genus Curculigo, as a folk herbal medicine, has been used for many years in China, treating impotence, limb limpness, and arthritis of the lumbar and knee joints. The last systematic review of the genus Curculigo was written in 2013, scientifically categorizing the phytochemistry and biological activities. Hitherto, the original compounds and their pharmacological activities were presented as the development of this genus, but there is not an updated review. To conclude the progression of the genus Curculigo, we collected the new literature published from 2013 to 2021 in PubMed, Web of Science, Google Scholar databases, and the Chinese National Knowledge Infrastructure. The novel chlorophenolic glucosides, curculigine, phenolic glycosides, orcinosides and polysaccharides were isolated from Curculigo. The new analyzing methods were established to control the quality of Curculigo as a herbal medicine. In addition, the pharmacological effects of Curculigo focused on anti-diabetes, antibacterial, anti-inflammatory, osteoporosis, antioxidation, etc. The antitumor and neuroprotective activities were newly explored in recent years. The application of herbal medicine was gradually developed in scientific methods. The medicinal value of the genus Curculigo needs to further investigate its pharmacological mechanisms. This new review offers more insights into the exploitation of the pharmacological value of the genus Curculigo.

De novo transcriptome analysis and comparative expression profiling of genes associated with the taste-modifying protein neoculin in Curculigo latifolia and Curculigo capitulata fruits.

BACKGROUND: Curculigo latifolia is a perennial plant endogenous to Southeast Asia whose fruits contain the taste-modifying protein neoculin, which binds to sweet receptors and makes sour fruits taste sweet. Although similar to snowdrop (Galanthus nivalis) agglutinin (GNA), which contains mannose-binding sites in its sequence and 3D structure, neoculin lacks such sites and has no lectin activity. Whether the fruits of C. latifolia and other Curculigo plants contain neoculin and/or GNA family members was unclear. RESULTS: Through de novo RNA-seq assembly of the fruits of C. latifolia and the related C. capitulata and detailed analysis of the expression patterns of neoculin and neoculin-like genes in both species, we assembled 85,697 transcripts from C. latifolia and 76,775 from C. capitulata using Trinity and annotated them using public databases. We identified 70,371 unigenes in C. latifolia and 63,704 in C. capitulata. In total, 38.6% of unigenes from C. latifolia and 42.6% from C. capitulata shared high similarity between the two species. We identified ten neoculin-related transcripts in C. latifolia and 15 in C. capitulata, encoding both the basic and acidic subunits of neoculin in both plants. We aligned these 25 transcripts and generated a phylogenetic tree. Many orthologs in the two species shared high similarity, despite the low number of common genes, suggesting that these genes likely existed before the two species diverged. The relative expression levels of these genes differed considerably between the two species: the transcripts per million (TPM) values of neoculin genes were 60 times higher in C. latifolia than in C. capitulata, whereas those of GNA family members were 15,000 times lower in C. latifolia than in C. capitulata. CONCLUSIONS: The genetic diversity of neoculin-related genes strongly suggests that neoculin genes underwent duplication during evolution. The marked differences in their expression profiles between C. latifolia and C. capitulata may be due to mutations in regions involved in transcriptional regulation. Comprehensive analysis of the genes expressed in the fruits of these two Curculigo species helped elucidate the origin of neoculin at the molecular level.

New chlorophenolic glycoside from Curculigo orchioides and their activities on 5α-reductase.

Two new chlorophenolic glucosides curculigines P (1) and Q (2), together with seven known compounds (3-9), were isolated from the dried rhizomes of Curculigo orchioides. Their structures were determined by spectroscopic methods including 1 D, 2 D NMR and MS. All the isolated compounds were evaluated on 5α-reductase activity by a HaCaT-based bioassay. Compounds 1-9 showed varying degrees of inhibiting activity against 5α-reductase, while compound 1 indicated the most potent inhibitory effect.[Formula: see text].

Inhibitory evaluation of Curculigo latifolia on α-glucosidase, DPP (IV) and in vitro studies in antidiabetic with molecular docking relevance to type 2 diabetes mellitus.

The inhibition of α-glucosidase and DPP enzymes capable of effectively reducing blood glucose level in the management of type 2 diabetes. The purpose of the present study is to evaluate the inhibitory potential of α-glucosidase and DPP (IV) activity including with the 2-NBDG uptake assay and insulin secretion activities through in vitro studies. The selected of active compounds obtained from the screening of compounds by LC-MS were docked with the targeted enzyme that involved in the mechanism of T2DM. From the results, root extracts displayed a better promising outcome in α-glucosidase (IC50 2.72 ± 0.32) as compared with the fruit extracts (IC50 3.87 ± 0.32). Besides, root extracts also displayed a better activity in the inhibition of DPP (IV), enhance insulin secretion and glucose uptake activity. Molecular docking results revealing that phlorizin binds strongly with α-glucosidase, DPP (IV) and Insulin receptor (IR) enzymes with achieving the lowest binding energy value. The present work suggests several of the compounds have the potential that contribute towards inhibiting α-glucosidase and DPP (IV) and thus effective in lowering post-prandial hyperglycaemia.

Prediction of Targets of Curculigoside A in Osteoporosis and Rheumatoid Arthritis Using Network Pharmacology and Experimental Verification.

PURPOSE: Network pharmacology is considered to be the next-generation drug development model that uses bioinformatics to predict and identify multiple drug targets and interactions in diseases. Here, network pharmacology was used to investigate the mechanism by which Curculigoside A (CA) acts in rheumatoid arthritis (RA) and osteoporosis. METHODS: First, TCMSP and SwissADME were applied to predict the druggability of CA. Then, potential targets were identified from overlapping data in SwissTarget and TargetNet, and targets were analyzed using Genemania and DAVID6.8 to obtain information about the GO and KEGG pathways. Ultimately, the drug-target-pathway network was identified after using Cytoscape 3.0 for visualization. Besides, qPCR was used to validate the predicted five major genes targets (EGFR, MAP2K1, MMP2, FGFR1, and MCL1). RESULTS: The results of TCMSP and SwissADME demonstrated that CA exhibits good druggability; 26 potential protein targets were classified by SwissTarget and TargetNet. The results of Genemania and DAVID6.8 indicated that CA probably caused anti-osteoporosis and anti-RA effects by regulating some biological pathways, especially nitrogen metabolism, estrogen signaling pathway, Rap1 signaling pathway, and PI3K/Akt signaling pathway. Besides, the result of Cytoscape 3.0 showed that the 26 targets participate in osteoporosis and RA-related pathways, metabolism, and other physiological processes. In vitro induced inflammation cell model experiments, the qPCR results showed that CA pretreatment significantly decreased the expression of EGFR, MAP2K1, MMP2, FGFR1, and MCL1 genes. CONCLUSION: These results suggested that network pharmacology may provide possible mechanism of how CA exerts therapeutic effects in osteoporosis and RA.

Structural Elucidation and Total Synthesis of Three 9-Norlignans from Curculigo capitulata.

Capitulactones A-C, three unprecedented 9-norlignans featuring a unique 3,5-dihydrofuro[2,3- d]oxepin-7(2 H)-one scaffold, were isolated from the roots of Curculigo capitulata. Their structures with absolute configurations were unambiguously established by a combination of spectroscopic data, ECD analysis, and total synthesis. Biomimetic total syntheses of three pairs of the corresponding enantiomers were achieved in 9-10 steps with overall yields of 14.8, 12.7, and 10.3%, respectively. Notably, the unique scaffold of the common western hemisphere of the molecules was constructed by using the oxidation-reduction strategy from benzodihydrofuran.

Structural elucidation and anti-osteoporosis activities of polysaccharides obtained from Curculigo orchioides.

Curculigo orchioides, is a traditional Chinese medicine, is used in strengthening tendons and bones. We evaluated the anti-osteoporosis activity of the crude polysaccharide (CO70) isolated from the rhizomes of C. orchioides in ovariectomized rats. CO70 showed excellent anti-osteoporosis activity comparable to that of 17ß-estradiol. To explore the constituents responsible for the anti-osteoporosis activity of CO70, a novel homogeneous heteropolysaccharide, COP70-3, was isolated and purified from CO70. COP70-3 has a main backbone chain of (1→5)-linked α-L-Araf, (1,3→5)-linked α-L-Araf, (1→6)-linked ß-D-Galp, (1→4)-linked ß-D-Manp, (1,2→5)-linked α-L-Araf, (1→3)-linked ß-L-Rhap, (1, 3→6)-linked ß-D-Manp, (1→3)-linked α-D-GalpA, (1,3→6)-linked ß-D-Galp and (1→6)-linked α-D-Glcp residues. Furthermore, 1.87 nM COP70-3 obviously promoted the differentiation of MC3T3-E1 cells, while 0.94 and 1.87 nM COP70-3 significantly improved the osteogenic mineralization rate. These data indicate that COP70-3 has favorable anti-osteoporosis activity in vitro.