OBJECTIVE Brain inflammation plays an important role in the pathophysiology of brain ischemicstroke,psychiatric and neurological diseases.During brain inflammation,microglia cells are activated and show pro-inflammato...OBJECTIVE Brain inflammation plays an important role in the pathophysiology of brain ischemicstroke,psychiatric and neurological diseases.During brain inflammation,microglia cells are activated and show pro-inflammatory M1 and anti-inflammatory M2 phenotypes,producing neurotoxic molecules and neurotrophic factors,respectively.We have previously discovered a novel natural product compound 3c exhibiting antiinflammatory effects in microglia cells,but the underlying mechanisms and its beneficial effects on brain inflammation and brain ischemia are unknown.METHODS The gene expression of M1 markers and M2 markers was measured by RT-PCR.The AMPK phosphorylation level and M2 marker CD206 protein expression were determined by Western blotting.TNFαrelease was measured by ELISA.The gene knowdown was performed by the si RNA transfection experiment.The LPS-induced brain inflammation mouse model and transient middle cerebral artery occlusion(t MCAO)stroke model were used.RESULTS We found that compound 3c inhibited M1polarization and promoted M2 polarization in LPS-stimulated BV2 and primary microglia cells,and these effects are mediated by Ca MKKβ/AMPK/JNK signaling pathway.Furthermore,compound 3c prevented M1 gene expression and enhanced M2 gene expression in a mouse model of LPS-induced neuroinflammation,and reduced the LPS-induced sickness behavior.In addition,compound 3c significantly reduced infarct volume,improved the neurological deficit,and reduced neuroinflammation in rats with acute focal cerebral ischemia.CONCLUSION Our results indicate that natural product compound 3c suppresses microglia activation by promoting M2 polarization and may provide a novel therapeutic approach to treat brain ischemic stroke associated with enhanced brain inflammation.展开更多
Several patient groups,particularly those with chronic illnesses such as cancer,cardiovascular disease,hepatitis C,and HIV/AIDS,often supplement their pharmacotherapeutic regimens with botanical natural products(NPs),...Several patient groups,particularly those with chronic illnesses such as cancer,cardiovascular disease,hepatitis C,and HIV/AIDS,often supplement their pharmacotherapeutic regimens with botanical natural products(NPs),raising concern for adverse NP-drug interactions.Like drug-drug interactions,common mechanisms underlying pharmacokinetic NP-drug interactions include induction and inhibition of drug metabolizing enzymes and transporters,leading toaltered systemic drug concentrations and potentially,suboptimal therapeutic effects.However,unlike for drug-drug interactions,rigorous guidelines for assessing the risk of NP-drug interactions are non-existent.Establishing such guidelines for NP-drug interactions poses challenges beyond those for drug-drug interactions because NPs are inherently complex mixtures that vary substantially in phytochemical composition.The National Center for Complementary and Integrative Health created the Center of Excellence for Natural Product-Drug Interaction(Na PDI)Research in September,2015.The mission of the Na PDI Center is to provide leadership in the identification,evaluation,and dissemination of potential clinically significant pharmacokinetic NP-drug interactions.A key deliverable of the Center is a set of Recommended Approaches to guide researchers in the proper conduct of NP-drug interaction studies.These approaches will be based on results generated from a series of Interaction Projects that will examine four methodically selected NPs as precipitants of metabolism-and/or transporter-mediated interactions with clinically relevant object drugs.Three of these NPs-green tea,goldenseal,and cannabinoids-have been advanced to Interaction Projects that include human mechanistic in vitro studies,physiologically-based pharmacokinetic modeling and simulation,and clinical studies.Key data generated from the Interaction Projects are being entered into a data repository,which will be disseminated to researchers via a public access portal.Collectively,the efforts of the Na PDI Center should lead to improved design of future NP-drug interaction research and ultimately,improved decisions on the optimal management of clinically relevant interactions.展开更多
OBJECTIVE The invasive sea lamprey(Petromyzon marinus)has devastated the ecosystem of the Laurentian Great Lakes.Application of pheromones to manipulate adult sea lamprey behavior is among the options considered for a...OBJECTIVE The invasive sea lamprey(Petromyzon marinus)has devastated the ecosystem of the Laurentian Great Lakes.Application of pheromones to manipulate adult sea lamprey behavior is among the options considered for alternative sea lamprey control techniques.The male sea lamprey sex pheromone is hypothesized to be possess multiple functions through actions of multiple components,some of which have yet to be characterized.Our objective is to isolate and characterize the bioactive components from water conditioned with sexually mature male sea lamprey.METHODS The water conditioned with sexually mature male sea lamprey was extracted by solid phase extraction and concentrated in vacuo.The compounds were isolated by liquid chromatography and elucidated by spectrometry and spectroscopy.Their biological activities were evaluated by electro-olfactogram recordings and two-choice maze behavioral assays.RESULTS Five novel bile salts,petromyzene A and B and petromyzone A-C,have been characterized.Petromyzene A and B featured either a unique,rearranged side chain or a rare cis-11,12-diol on the steroidal B-ring.Petromyzone A-C represented three novel highly oxidized sulfated bile alcohols possessing different hydroxylation,oxidation,and double bond patterns,which exemplify the chemical diversity of bile salts.These five bile salts were potent odorants that stimulated the adult sea lamprey olfactory epithelium in a concentration dependent manner and showed detection thresholds between 10–13mol·L^(-1) and 10^(–11)mol·L^(-1)(paired t-test,P<0.05).Experiments in the two-choice maze showed that all isolated compounds induced behavioral responses in ovulated females.CONCLUSION The five novel compounds are likely additional components of pheromones released by sexually mature male sea lamprey,and may provide useful behavioral manipulation tools to be implemented with the integrated management of the destructive and invasive sea lamprey in the Laurentian Great Lakes.展开更多
Countless studies have been devoted to the scientific evaluation of the safety and/or efficacy of botanical natural products.Investigators involved in such studies face a unique set of challenges.Natural products diff...Countless studies have been devoted to the scientific evaluation of the safety and/or efficacy of botanical natural products.Investigators involved in such studies face a unique set of challenges.Natural products differ from their pharmaceutical counterparts in that they are typically complex mixtures,for which the identities and quantities of components present are not known.To further complicate matters,the composition of these mixtures will vary depending on source material and method of preparation.Investigators conducting clinical trials with complex botanical natural products must choose from a myriad of potential preparations,which may vary greatly in composition.In making such decisions,it is extremely useful to know which components of the mixture are most likely to be responsible for its purported biological activity(the"active constituents").The gold standard approach for identifying active constituents of botanical natural products is bioassay-guided fractionation,in which the mixture is subjected to successive rounds of purification and bioassays until an active compound is identified.Bioassay guided fractionation has historically played a critical role in drug discovery,but is,nonetheless,fraught with challenges.The process is biased towards the most abundant and easily isolatable mixture components,which may not be the most biologically active.Furthermore,if multiple compounds contribute either additively,antagonistically,or synergistically to the observed biological activity of the mixture,activity may be lost upon isolation.As a complementary strategy to bioassay-guided fractionation,our research group has developed untargeted metabolomics strategies to aid in the identification of bioactive mixture components.These strategies involve profiling botanical mixtures using ultraperformance chromatography coupled to high resolving power mass spectrometry.The resulting chemical data is then integrated with biological assay data using biochemometric data analysis strategies.Several case studies will be presented illustrating how this approach can be applied,including for the identification of compounds from the botanical green(Camellia sinensis)that inhibit drug metabolizing enzymes.Such studies are being conducted as part of the Center for Excellence in Natural Product Drug Interaction Studies(Na PDI),which is supported by a cooperative agreement with the National Center for Complementary and Integrative Health,a component of the National Institutes of Health.展开更多
文摘OBJECTIVE Brain inflammation plays an important role in the pathophysiology of brain ischemicstroke,psychiatric and neurological diseases.During brain inflammation,microglia cells are activated and show pro-inflammatory M1 and anti-inflammatory M2 phenotypes,producing neurotoxic molecules and neurotrophic factors,respectively.We have previously discovered a novel natural product compound 3c exhibiting antiinflammatory effects in microglia cells,but the underlying mechanisms and its beneficial effects on brain inflammation and brain ischemia are unknown.METHODS The gene expression of M1 markers and M2 markers was measured by RT-PCR.The AMPK phosphorylation level and M2 marker CD206 protein expression were determined by Western blotting.TNFαrelease was measured by ELISA.The gene knowdown was performed by the si RNA transfection experiment.The LPS-induced brain inflammation mouse model and transient middle cerebral artery occlusion(t MCAO)stroke model were used.RESULTS We found that compound 3c inhibited M1polarization and promoted M2 polarization in LPS-stimulated BV2 and primary microglia cells,and these effects are mediated by Ca MKKβ/AMPK/JNK signaling pathway.Furthermore,compound 3c prevented M1 gene expression and enhanced M2 gene expression in a mouse model of LPS-induced neuroinflammation,and reduced the LPS-induced sickness behavior.In addition,compound 3c significantly reduced infarct volume,improved the neurological deficit,and reduced neuroinflammation in rats with acute focal cerebral ischemia.CONCLUSION Our results indicate that natural product compound 3c suppresses microglia activation by promoting M2 polarization and may provide a novel therapeutic approach to treat brain ischemic stroke associated with enhanced brain inflammation.
文摘Several patient groups,particularly those with chronic illnesses such as cancer,cardiovascular disease,hepatitis C,and HIV/AIDS,often supplement their pharmacotherapeutic regimens with botanical natural products(NPs),raising concern for adverse NP-drug interactions.Like drug-drug interactions,common mechanisms underlying pharmacokinetic NP-drug interactions include induction and inhibition of drug metabolizing enzymes and transporters,leading toaltered systemic drug concentrations and potentially,suboptimal therapeutic effects.However,unlike for drug-drug interactions,rigorous guidelines for assessing the risk of NP-drug interactions are non-existent.Establishing such guidelines for NP-drug interactions poses challenges beyond those for drug-drug interactions because NPs are inherently complex mixtures that vary substantially in phytochemical composition.The National Center for Complementary and Integrative Health created the Center of Excellence for Natural Product-Drug Interaction(Na PDI)Research in September,2015.The mission of the Na PDI Center is to provide leadership in the identification,evaluation,and dissemination of potential clinically significant pharmacokinetic NP-drug interactions.A key deliverable of the Center is a set of Recommended Approaches to guide researchers in the proper conduct of NP-drug interaction studies.These approaches will be based on results generated from a series of Interaction Projects that will examine four methodically selected NPs as precipitants of metabolism-and/or transporter-mediated interactions with clinically relevant object drugs.Three of these NPs-green tea,goldenseal,and cannabinoids-have been advanced to Interaction Projects that include human mechanistic in vitro studies,physiologically-based pharmacokinetic modeling and simulation,and clinical studies.Key data generated from the Interaction Projects are being entered into a data repository,which will be disseminated to researchers via a public access portal.Collectively,the efforts of the Na PDI Center should lead to improved design of future NP-drug interaction research and ultimately,improved decisions on the optimal management of clinically relevant interactions.
文摘OBJECTIVE The invasive sea lamprey(Petromyzon marinus)has devastated the ecosystem of the Laurentian Great Lakes.Application of pheromones to manipulate adult sea lamprey behavior is among the options considered for alternative sea lamprey control techniques.The male sea lamprey sex pheromone is hypothesized to be possess multiple functions through actions of multiple components,some of which have yet to be characterized.Our objective is to isolate and characterize the bioactive components from water conditioned with sexually mature male sea lamprey.METHODS The water conditioned with sexually mature male sea lamprey was extracted by solid phase extraction and concentrated in vacuo.The compounds were isolated by liquid chromatography and elucidated by spectrometry and spectroscopy.Their biological activities were evaluated by electro-olfactogram recordings and two-choice maze behavioral assays.RESULTS Five novel bile salts,petromyzene A and B and petromyzone A-C,have been characterized.Petromyzene A and B featured either a unique,rearranged side chain or a rare cis-11,12-diol on the steroidal B-ring.Petromyzone A-C represented three novel highly oxidized sulfated bile alcohols possessing different hydroxylation,oxidation,and double bond patterns,which exemplify the chemical diversity of bile salts.These five bile salts were potent odorants that stimulated the adult sea lamprey olfactory epithelium in a concentration dependent manner and showed detection thresholds between 10–13mol·L^(-1) and 10^(–11)mol·L^(-1)(paired t-test,P<0.05).Experiments in the two-choice maze showed that all isolated compounds induced behavioral responses in ovulated females.CONCLUSION The five novel compounds are likely additional components of pheromones released by sexually mature male sea lamprey,and may provide useful behavioral manipulation tools to be implemented with the integrated management of the destructive and invasive sea lamprey in the Laurentian Great Lakes.
文摘Countless studies have been devoted to the scientific evaluation of the safety and/or efficacy of botanical natural products.Investigators involved in such studies face a unique set of challenges.Natural products differ from their pharmaceutical counterparts in that they are typically complex mixtures,for which the identities and quantities of components present are not known.To further complicate matters,the composition of these mixtures will vary depending on source material and method of preparation.Investigators conducting clinical trials with complex botanical natural products must choose from a myriad of potential preparations,which may vary greatly in composition.In making such decisions,it is extremely useful to know which components of the mixture are most likely to be responsible for its purported biological activity(the"active constituents").The gold standard approach for identifying active constituents of botanical natural products is bioassay-guided fractionation,in which the mixture is subjected to successive rounds of purification and bioassays until an active compound is identified.Bioassay guided fractionation has historically played a critical role in drug discovery,but is,nonetheless,fraught with challenges.The process is biased towards the most abundant and easily isolatable mixture components,which may not be the most biologically active.Furthermore,if multiple compounds contribute either additively,antagonistically,or synergistically to the observed biological activity of the mixture,activity may be lost upon isolation.As a complementary strategy to bioassay-guided fractionation,our research group has developed untargeted metabolomics strategies to aid in the identification of bioactive mixture components.These strategies involve profiling botanical mixtures using ultraperformance chromatography coupled to high resolving power mass spectrometry.The resulting chemical data is then integrated with biological assay data using biochemometric data analysis strategies.Several case studies will be presented illustrating how this approach can be applied,including for the identification of compounds from the botanical green(Camellia sinensis)that inhibit drug metabolizing enzymes.Such studies are being conducted as part of the Center for Excellence in Natural Product Drug Interaction Studies(Na PDI),which is supported by a cooperative agreement with the National Center for Complementary and Integrative Health,a component of the National Institutes of Health.
