2-Butoxytetrahydrofuran, Isolated from Holothuria scabra, Attenuates Aggregative and Oxidative Properties of α-Synuclein and Alleviates Its Toxicity in a Transgenic Caenorhabditis elegans Model of Parkinson's Disease.

Aggregative α-synuclein and incurring oxidative stress are pivotal cascading events, leading to dopaminergic (DAergic) neuronal loss and contributing to clinical manifestations of Parkinson's disease (PD). Our previous study demonstrated that 2-butoxytetrahydrofuran (2-BTHF), isolated from Holothuria scabra (H. scabra), could inhibit amyloid-ß aggregation and its ensuing toxicity, which leads to Alzheimer's disease. In the present study, we found that 2-BTHF also attenuated the aggregative and oxidative activities of α-synuclein and lessened its toxicity in a transgenic Caenorhabditis elegans (C. elegans) PD model. Such worms treated with 100 µM of 2-BTHF showed substantial reductions in α-synuclein accumulation and DAergic neurodegeneration. Mechanistically, 2-BTHF, at this concentration, significantly decreased aggregation of monomeric α-synuclein and restored locomotion and dopamine-dependent behaviors. Molecular docking exhibited potential bindings of 2-BTHF to HSF-1 and DAF-16 transcription factors. Additionally, 2-BTHF significantly increased the mRNA transcripts of genes encoding proteins involved in proteostasis, including the molecular chaperones hsp-16.2 and hsp-16.49, the ubiquitination/SUMOylation-related ubc-9 gene, and the autophagy-related genes atg-7 and lgg-1. Transcriptomic profiling revealed an additional mechanism of 2-BTHF in α-synuclein-expressing worms, which showed upregulation of PPAR signaling cascades that mediated fatty acid metabolism. 2-BTHF significantly restored lipid deposition, upregulated the fat-7 gene, and enhanced gcs-1-mediated glutathione synthesis in the C. elegans PD model. Taken together, this study demonstrated that 2-BTHF could abrogate aggregative and oxidative properties of α-synuclein and attenuate its toxicity, thus providing a possible therapeutic application for the treatment of α-synuclein-induced PD.

Holothuria scabra extracts confer neuroprotective effect in C. elegans model of Alzheimer's disease by attenuating amyloid-ß aggregation and toxicity.

Background and aim: Alzheimer's disease (AD) is the most common aged-related neurodegenerative disorder that is associated with the toxic amyloid-ß (Aß) aggregation in the brain. While the efficacies of available drugs against AD are still limited, natural products have been shown to possess neuroprotective potential for prevention and therapy of AD. This study aimed to investigate the neuroprotective effects of H. scabra extracts against Aß aggregation and proteotoxicity in C. elegans model of Alzheimer's diseases. Experimental procedure: Whole bodies (WB) and body wall (BW) of H. scabra were extracted and fractionated into ethyl acetate (WBEA, BWEA), butanol (WBBU, BWBU), and ethanol (BWET). Then C. elegans AD models were treated with these fractions and investigated for Aß aggregation and polymerization, biochemical and behavioral changes, and level of oxidative stress, as well as lifespan extension. Results and conclusion: C. elegans AD model treated with H. scabra extracts, especially triterpene glycoside-rich ethyl acetate and butanol fractions, exhibited significant reduction of Aß deposition. These H. scabra extracts also attenuated the paralysis behavior and improved the neurological defects in chemotaxis caused by Aß aggregation. Immunoblot analysis revealed decreased level of Aß oligomeric forms and the increased level of Aß monomers after treatments with H. scabra extracts. In addition, H. scabra extracts reduced reactive oxygen species and increased the mean lifespan of the treated AD worms. In conclusion, this study demonstrated strong evidence of anti-Alzheimer effects by H. scabra extracts, implying that these extracts can potentially be applied as natural preventive and therapeutic agents for AD. Taxonomy classification by EVISE: Alzheimer's disease, Neurodegenerative disorder, Traditional medicine, Experimental model systems, Molecular biology.

Neuroprotective effects of a medium chain fatty acid, decanoic acid, isolated from H. leucospilota against Parkinsonism in C. elegans PD model.

Sea cucumbers are marine organism that have long been used for food and traditional medicine in Asian countries. Recently, we have shown that ethyl acetate fraction (HLEA) of the crude extract of the black sea cucumber, Holothuria leucospilota, could alleviate Parkinsonism in Caenorhabditis elegans PD models. In this study, we found that the effective neuroprotective activity is attributed to HLEA-P1 compound chemically isolated and identified in H. leucospilota ethyl acetate. We reported here that HLEA-P1 could attenuate DAergic neurodegeneration, improve DAergic-dependent behaviors, reduce oxidative stress in 6-OHDA-induced C. elegans. In addition, HLEA-P1 reduced α-synuclein aggregation, improved behavior deficit and recovered lipid deposition in transgenic C. elegans overexpressing α-synuclein. We also found that HLEA-P1 activates nuclear localization of DAF-16 transcription factor of insulin/IGF-1 signaling (IIS) pathway. Treatment with 25 µg/ml of HLEA-P1 upregulated transcriptional activity of DAF-16 target genes including anti-oxidant genes (such as sod-3) and small heat shock proteins (such as hsp16.1, hsp16.2, and hsp12.6) in 6-OHDA-induced worms. In α-synuclein-overexpressed C. elegans strain, treatment with 5 µg/ml of HLEA-P1 significantly activated mRNA expression of sod-3 and hsp16.2. Chemical analysis demonstrated that HLEA-P1 compound is decanoic acid/capric acid. Taken together, our findings revealed that decanoic acid isolated from H. leucospilota exerts anti-Parkinson effect in C. elegans PD models by partly modulating IIS/DAF-16 pathway.

Neurorescue Effects of Frondoside A and Ginsenoside Rg3 in C. elegans Model of Parkinson's Disease.

Parkinson's disease (PD) is a currently incurable neurodegenerative disorder characterized by the loss of dopaminergic (DAergic) neurons in the substantia nigra pars compacta and α-synuclein aggregation. Accumulated evidence indicates that the saponins, especially from ginseng, have neuroprotective effects against neurodegenerative disorders. Interestingly, saponin can also be found in marine organisms such as the sea cucumber, but little is known about its effect in neurodegenerative disease, including PD. In this study, we investigated the anti-Parkinson effects of frondoside A (FA) from Cucumaria frondosa and ginsenoside Rg3 (Rg3) from Panax notoginseng in C. elegans PD model. Both saponins were tested for toxicity and optimal concentration by food clearance assay and used to treat 6-OHDA-induced BZ555 and transgenic α-synuclein NL5901 strains in C. elegans. Treatment with FA and Rg3 significantly attenuated DAergic neurodegeneration induced by 6-OHDA in BZ555 strain, improved basal slowing rate, and prolonged lifespan in the 6-OHDA-induced wild-type strain with downregulation of the apoptosis mediators, egl-1 and ced-3, and upregulation of sod-3 and cat-2. Interestingly, only FA reduced α-synuclein aggregation, rescued lifespan in NL5901, and upregulated the protein degradation regulators, including ubh-4, hsf-1, hsp-16.1 and hsp-16.2. This study indicates that both FA and Rg3 possess beneficial effects in rescuing DAergic neurodegeneration in the 6-OHDA-induced C. elegans model through suppressing apoptosis mediators and stimulating antioxidant enzymes. In addition, FA could attenuate α-synuclein aggregation through the protein degradation process.

Diterpene glycosides from Holothuria scabra exert the α-synuclein degradation and neuroprotection against α-synuclein-Mediated neurodegeneration in C. elegans model.

ETHNOPHARMACOLOGICAL RELEVANCE: Holothuria (Metriatyla) scabra Jaeger (H. scabra), sea cucumber, is the marine organism that has been used as traditional food and medicine to gain the health benefits since ancient time. Although our recent studies have shown that crude extracts from H. scabra exhibited neuroprotective effects against Parkinson's disease (PD), the underlying mechanisms and bioactive compounds are still unknown. AIM OF THE STUDY: In the present study, we examined the efficacy of purified compounds from H. scabra and their underlying mechanism on α-synuclein degradation and neuroprotection against α-synuclein-mediated neurodegeneration in a transgenic Caenorhabditis elegans PD model. MATERIAL AND METHODS: The H. scabra compounds (HSEA-P1 and P2) were purified and examined for their toxicity and optimal dose-range by food-clearance and lifespan assays. The α-synuclein degradation and neuroprotection against α-synuclein-mediated neurodegeneration were determined using transgenic C. elegans model, Punc-54::α-syn and Pdat-1:: α-syn; Pdat-1::GFP, respectively, and then further investigated by determining the behavioral assays including locomotion rate, basal slowing rate, ethanol avoidance, and area-restricted searching. The underlying mechanisms related to autophagy were clarified by quantitative PCR and RNAi experiments. RESULTS: Our results showed that HSEA-P1 and HSEA-P2 significantly diminished α-synuclein accumulation, improved motility deficits, and recovered the shortened lifespan. Moreover, HSEA-P1 and HSEA-P2 significantly protected dopaminergic neurons from α-synuclein toxicity and alleviated dopamine-associated behavioral deficits, i.e., basal slowing, ethanol avoidance, and area-restricted searching. HSEA-P1 and HSEA-P2 also up-regulated autophagy-related genes, including beclin-1/bec-1, lc-3/lgg-1, and atg-7/atg-7. RNA interference (RNAi) of these genes in transgenic α-synuclein worms confirmed that lc-3/lgg-1 and atg-7/atg-7 were required for α-synuclein degradation and DAergic neuroprotection activities of HSEA-P1 and HSEA-P2. NMR and mass spectrometry analysis revealed that the HSEA-P1 and HSEA-P2 contained diterpene glycosides. CONCLUSION: These findings indicate that diterpene glycosides extracted from H. scabra decreases α-synuclein accumulation and protects α-synuclein-mediated DAergic neuronal loss and its toxicities via lgg-1 and atg-7.

Downregulation of eEF1A/EFT3-4 Enhances Dopaminergic Neurodegeneration After 6-OHDA Exposure in C. elegans Model.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the aggregation of α-synuclein protein and selective death of dopaminergic (DA) neurons in the substantia nigra of the midbrain. Although the molecular pathogenesis of PD is not completely understood, a recent study has reported that eukaryotic translation elongation factor 1 alpha (eEF1A) declined in the PD-affected brain. Therefore, the roles of eEF1A1 and eEF1A2 in the prevention of DA neuronal cell death in PD are aimed to be investigated. Herein, by using Caenorhabditis elegans as a PD model, we investigated the role of eft-3/eft-4, the worm homolog of eEF1A1/eEF1A2, on 6-hydroxydopamine (6-OHDA)-induced DA neuron degeneration. Our results demonstrated that the expressions of eft-3 and eft-4 were decreased in the 6-OHDA-induced worms. RNA interference (RNAi) of eft-3 and eft-4 resulted in dramatic exacerbation of DA neurodegeneration induced by 6-OHDA, as well as aggravated the food-sensing behavior, ethanol avoidance, and decreased lifespan when compared with only 6-OHDA-induced worms. Moreover, downregulation of eft-3/4 in 6-OHDA-induced worms suppressed the expression of the anti-apoptotic genes, including PI3K/age-1, PDK-1/pdk-1, mTOR/let-363, and AKT-1,2/akt-1,2, promoting the expression of apoptotic genes such as BH3/egl-1 and Caspase-9/ced-3. Collectively, these findings indicate that eEF1A plays an important role in the 6-OHDA-induced neurodegeneration through the phosphatidylinositol 3-kinase (PI3K)/serine/threonine protein kinase (Akt)/mammalian target of rapamycin (mTOR) pathway and that eEF1A isoforms may be a novel and effective pro-survival factor in protective DA neurons against toxin-induced neuronal death.

Saponin-enriched extracts from body wall and Cuvierian tubule of Holothuria leucospilota reduce fat accumulation and suppress lipogenesis in Caenorhabditis elegans.

BACKGROUND: Saponins have been shown to possess many pharmacological properties, including altered fat metabolism. The black sea cucumber, Holothuria leucospilota, is a marine animal that contains a specialized organ called a Cuvierian tubule that produces and secrete the bioactive saponins into the tubules and body wall. Therefore, the aims of this study are to investigate the anti-obesity effect of saponins extracted from body wall and Cuvierian tubules of H. leucospilota. RESULTS: The butanol extracts of H. leucospilota body wall and Cuvierian tubules containing high amounts of saponins significantly reduced fat deposition and triglyceride levels in Caenorhabditis elegans fed with 50 mmol L-1 glucose. Moreover, the saponin-enriched extracts of H. leucospilota significantly restored the lifespan of 2% glucose-fed worms (18.71%). Green fluorescence protein-labeled sbp-1 gene expression and nuclear translocation of daf-16 were also significantly decreased in H. leucospilota treatment. The saponin-enriched extracts downregulated the messenger RNA expressions of genes involved in fat storage and metabolism, including sbp-1, cebp, and daf-16 but upregulated the expression of nhr-49 gene. CONCLUSION: Our results suggest that H. leucospilota-derived saponins may mediate the reduction of glucose-induced fat accumulation through sbp-1, cebp, daf-16 and nhr-9 pathways. Therefore, the H. leucospilota extracts could be used as nutraceuticals for anti-obesity prevention. © 2019 Society of Chemical Industry.

Anti-Parkinson activity of bioactive substances extracted from Holothuria leucospilota.

Parkinson's disease (PD) is a well-known neurodegenerative disorder characterized by dopaminergic (DA) neuron loss and α-synuclein aggregation. Recent study revealed that the extracts from sea cucumber, Holothuroidea spp., exhibited neuroprotective and lifespan extension effects in Caenorhabditis elegans model. Interestingly, the black sea cucumber, Holothuria leucospilota, possesses body wall and a specialized organ called cuvierian tubules containing high amount of bioactive compounds. In this study, the neuroprotective effects of the body wall (BW) and cuvierian tubules (CT) from this sea cucumber against PD were evaluated using C. elegans as a model. H. leucospilota were extracted using ethanol (ET), ethyl acetate (EA), butanol (BU) and aqueous (AQ) fractions. Extracts from these fractions were used to treat the 6-OHDA-induced BZ555 and α-synuclein expressing NL5901 strains of C. elegans. Treatment with ET, EA, BU and AQ fractions of H. leucospilota extracts could significantly prevent degeneration of DA neurons in 6-OHDA-induced worms, improve food-sensing behavior mediated by DA neurons, and up-regulate cat-2 and sod-3 gene expressions. These results indicate the neuroprotective activity of the extracts which may be attributed to the anti-oxidant activity of the bioactive compounds. Moreover, α-synuclein aggregation was significantly reduced together with the recovery of lipid deposition upon the treatment with H. leucospilota extracts. In addition, treatment with H. leucospilota extracts was able to increase the lifespan of 6-OHDA-induced N2. NMR analysis revealed the major chemical components in the effective EA fractions were terpenoids, steroids, saponins, and glycosides. In summary, H. leucospilota extracts exhibited anti-Parkinson effect in both toxin-induced and transgenic C. elegans models of PD. Further study will be performed to elucidate the most effective anti-PD molecules which will lead to the development of anti-PD drug.

Holothuria scabra extracts possess anti-oxidant activity and promote stress resistance and lifespan extension in Caenorhabditis elegans.

Holothuria scabra is a sea cucumber that is mostly found in the Indo-Pacific region including Thailand. Extracts from many sea cucumbers possess pharmacological activities proposed to benefit human health. In this study, we investigated the anti-oxidant and anti-ageing activities of extracts from H. scabra by using Caenorhabditis elegans as a model organism. Parts of H. scabra were solvent-extracted and divided into nine fractions including whole body-hexane (WBHE), whole body-ethyl acetate (WBEA), whole body-butanol (WBBU), body wall-hexane (BWHE), body wall-ethyl acetate (BWEA), body wall-butanol (BWBU), viscera-hexane (VIHE), viscera-ethyl acetate (VIEA), and viscera-butanol (VIBU). All fractions of the extracts were tested for anti-oxidant activities by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays and for anti-ageing effects by lifespan assays using C. elegans as a model. The results showed anti-oxidant properties in all fractions with the highest activity shown by the DPPH assay in WBBU (EC50 = 3.12 ±â€¯0.09 mg/ml), and by the ABTS assay in WBHE (EC50 = 0.31 ±â€¯0.10 mg/ml). In lifespan assays the highest anti-ageing effect was detected in WBBU- and BWEA-treated C. elegans with increased mean lifespans of 8.12% and 4.77%, respectively. Furthermore, WBBU and BWEA-treated C. elegans exhibited significantly higher resistance against heat shock and paraquat-induced oxidative stresses than controls. By using LC-MS/MS, both extracts were characterized to contain triterpene glycosides as the main bioactive components. To explore mechanisms of H. scabra extracts on longevity and stress resistance, worms with genetic mutations in anti-ageing pathways were analyzed and showed that WBBU and BWEA did not prolong the lifespan of daf-16, age-1, sir-2.1, jnk-1, sek-1, and osr-1 mutants, suggesting that these genetic pathways are involved in mediating the anti-ageing effects of the H. scabra extracts. Moreover, WBBU and BWEA enhanced the nuclear translocation of the FoxO/DAF-16 transcription factor, and increased mRNA expression of this gene and its downstream targets sod-3, hsp12.3, and hsp16.2. In conclusion, this study strongly demonstrates anti-oxidant and anti-ageing properties of H. scabra extracts containing triterpene glycosides, which, in the C. elegans model, may be mediated via the insulin/IGF-1 signaling (IIS)-DAF-16 pathway.

Holothuria scabra extracts exhibit anti-Parkinson potential in C. elegans: A model for anti-Parkinson testing.

OBJECTIVES: Parkinson's disease (PD) is associated with aggregation of α-synuclein and selective death of dopaminergic (DA) neurons in the substantia nigra, thereby leading to cognitive and motor impairments. Nowadays, the drugs commonly used for PD treatment, such as levodopa, provide only symptomatic relief. Therefore, seeking new drugs against PD, especially from plants and marine organisms, is one of the major research areas to be explored. This study aimed to investigate the anti-Parkinson activity of the extracts from the sea cucumber, Holothuria scabra, by using Caenorhabditis elegans as a model. METHODS: H. scabra was solvent-extracted and subdivided into six fractions including whole body-ethyl acetate (WBEA), body wall-ethyl acetate (BWEA), viscera-ethyl acetate (VIEA), whole body-butanol (WBBU), body wall-butanol (BWBU), and viscera-butanol (VIBU). The extracts were tested in C. elegans BZ555 strain expressing the green fluorescent protein (GFP) specifically in the DA neurons and NL5901 strain expressing human α-synuclein in the muscle cells. RESULTS: WBEA, BWEA, and WBBU fractions of H. scabra extracts at 500 µg/ml significantly attenuated DA neuron-degeneration induced by selective cathecholamine neurotoxin 6-hydroxydopamine (6-OHDA) in the BZ555 strain. Moreover, the extracts also reduced α-synuclein aggregation and restored lipid content in NL5901, as well as improved food-sensing behavior and prolonged lifespan in the 6-OHDA-treated wild-type strain. DISCUSSION: The study indicated that the H. scabra extracts have anti-Parkinson potential in the C. elegans model. These findings encourage further investigations on using the H. scabra extract, as well as its active constituent compounds, as a possible preventive and/or therapeutic intervention against PD.

Electrospun cellulose acetate doped with astaxanthin derivatives from Haematococcus pluvialis for in vivo anti-aging activity.

This research aims to study the release, in vivo anti-aging activity against Caenorhabditis elegans and stability of astaxanthins in a crude acetone extract of Haematococcus pluvialis from electrospun cellulose acetate (CA) nanofibers. The content and 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging activity of astaxanthins in the crude extract were also determined. The content of astaxanthins was reported in terms of total carotenoid content (TCC) and found to be 10.75 ± 0.16 mg gcE-1. IC50 of DPPH radical scavenging activity for astaxanthins was 233.33 ± 4.18 µg mL-1. It has been well known that astaxanthins are very unstable under environmental conditions, so the electrospinning technique was used to enhance their stability. In order to fabricate CA nanofibers containing a crude acetone extract of H. pluvialis, various solvent systems and percent loading of the crude acetone extract were studied. The optimal solvent system for fabrication of CA nanofibers was the acetone/dimethylformamide (DMF) system (2 : 1 v/v) with incorporation of 0.25% v/v Tween80, resulting in good morphology of CA nanofibers with av. 420 nm diameter. The loading efficiency (%) of the crude astaxanthins extract was 5% w/w of CA. With regard to the results of the in vivo oxidative stress assay, C. elegans pre-treated with 200 µg mL-1 of the crude extract had a survival percent of 56 after administration of 250 mM of paraquat for 8 h. Under phosphate-buffered saline (pH 7.4) containing 10% v/v acetone, the release of astaxanthins from the CA nanofibers loaded with the crude extract exhibited a prolonged profile. The stability of astaxanthins in electrospun CA nanofibers was examined using the freeze-thaw cycle testing through a DPPH radical scavenging assay. It was found that their stability was significantly different (P < 0.05) after the 12th freeze-thaw cycle compared with the crude extract.