摘要
Antrodia cinnamomea 牛樟芝(Syn.Antrodia camphorata and Taiwanofungus camphorata) is a precious edible fungus endemic to Taiwan that has long been used as a folk remedy for treating various diseases including liver diseases,hypertension,abdominal pain,and cancer.To investigate the antiinflammation activity of A.cinnampmea,a lipopolysaccaride(LPS)-challenged ICR mouse acute inflammation model and a LPS-induced macrophage model were used.Ethanol extract of A.cinnampmea significantly inhibited expression of i NOS and COX-2 in the liver of LPS-challenged acute inflammatory mice.The ethyl acetate fraction and its isolated compound,antrocamphin A,significantly suppressed nitrite/nitrate concentration in LPS-challenged RAW 264.7 cells.Antrocamphin A showed potent anti-inflammatory activity by suppressing pro-inflammatory molecule release via the down-regulation of i NOS and COX-2 expression through the NF-κB pathway.This study,therefore,first demonstrates the bioactive compound of A.cinnampmea and illustrates the mechanism by which it confers its anti-inflammatory activity(Hsieh et al.,2010).Besides the antiinflammation activity study,the hepatoprotective efficacy of antroquinonol and ethanolic extracts of mycelia of A.cinnamomea(EMAC) in vitro and in vivo by us.Based on the results obtained in our study,antroquinonol pretreatment significantly inhibited ethanol-induced AST,ALT,ROS,NO,MDA production and GSH depletion in Hep G2 cells.Western blot and RT-PCR analysis showed that antroquinonol enhanced Nrf-2 activation and its downstream antioxidant gene HO-1 via MAPK pathway.This mechanism was then confirmed in vivo in an acute ethanol intoxicated mouse model:serum ALT and AST production,hepatocellular lipid peroxidation and GSH depletion was prevented by EMAC in a dose-dependent manner.EMAC significantly enhanced HO-1 and Nrf-2 activation via MAPKs consistent with in vitro studies.Ethanol-induced hepatic swelling and hydropic degeneration of hepatocytes was significantly inhibited by EMAC in a dose-dependent manner.These results provide a scientific basis for the hepatoprotective effects of A.cinnamomea.Data also imply that antroquinonol,a potent bioactive compound may be responsible for the hepatoprotective activity of A.cinnamomea.This study highly supported our traditional knowledge that A.cinnamomea as a potential candidate for the treatment of alcoholic liver diseases(Senthil Kumar et al.,2011).On the other hand,an index of 13 representative metabolites from the ethanol extract of A.cinnamomea fruiting body was established for use in quality evaluation.Most of the index compounds selected,particularly the ergostane-type triterpenoids and polyacetylenes,possess good antiinflammation activity.A comparison of the metabolite profiles of different ethanol extracts from A.cinnamomea strains showed similar metabolites when the strains were grown on the original host wood(Cinnamomum kanehirai) and harvested after the same culture time period(9 months).Furthermore,the amounts of typical ergostane-type triterpenoids in A.cinnamomea increased with culture age.Culture substrates also influenced metabolite synthesis;with the same culture age,A.cinnamomea grown on the original host wood produced a richer array of metabolites than A.cinnamomea cultured on other wood species.We conclude that analysis of a fixed group of compounds including triterpenoids,benzolics,and polyacetylenes constitutes a suitable,reliable system to evaluate the quality of ethanol extract from A.cinnamomea fruiting bodies.The evaluation system established in this study may provide a platform for analysisof the products of A.cinnamomea(Lin et al.,2011).Besides fruiting bodies,we selected an A.cinnamomea mycelium health food product,which was produced by solid-state culture and certificate by Taiwan's Department of Health as "National Health Food",as the target for investigation.14 representative metabolites of A.cinnamomea mycelium(EMAC) were selected as index compounds to establish the metabolite profile for evaluation of EMAC product quality.It was also demonstrated that EMAC administration significantly reduced liver inflammation and serum oxidative stress in vivo.4-Acetylantroquinonol B obtained by a bioactivity-guided fractionation from EMAC was able to not only inhibit LPS-induced nitric oxide formation in macrophages but also protect against ethanol-induced oxidative stress in liver cells.The results suggest this A.cinnamomea product might be a potent antioxidative and anti-inflammatory supplement for chemoprevention(Wang et al.,2013).Recently,we found that Antrodin C(ADC),a maleimide derivative isolated from A.cinnamomea health food product inhibits TGF-β1-induced Epithelial-to-mesenchymal transition(EMT) and breast cancer cell metastasis in vitro.Our data suggested that ADC attenuates the TGF-β1-induced EMT,migration and invasion of human breast carcinoma through the suppression of Smad2/3 and β-catenin signaling pathways(Senthil Kumar et al.,2015).The similar activity was also approved for 2,3,5-trimethoxy-4-cresol(TMC),which was isolated from solid-state cultured mycelium of A.cinnamomea(strain no.LEACS-002).We demonstrated that TMC effectively suppresses movement,migration and invasion of lung cancer cells,and achieves an anti-cancer metastasis effect(Lin et al.,2015).
Antrodia cinnamomea 牛樟芝(Syn.Antrodia camphorata and Taiwanofungus camphorata) is a precious edible fungus endemic to Taiwan that has long been used as a folk remedy for treating various diseases including liver diseases,hypertension,abdominal pain,and cancer.To investigate the antiinflammation activity of A.cinnampmea,a lipopolysaccaride(LPS)-challenged ICR mouse acute inflammation model and a LPS-induced macrophage model were used.Ethanol extract of A.cinnampmea significantly inhibited expression of i NOS and COX-2 in the liver of LPS-challenged acute inflammatory mice.The ethyl acetate fraction and its isolated compound,antrocamphin A,significantly suppressed nitrite/nitrate concentration in LPS-challenged RAW 264.7 cells.Antrocamphin A showed potent anti-inflammatory activity by suppressing pro-inflammatory molecule release via the down-regulation of i NOS and COX-2 expression through the NF-κB pathway.This study,therefore,first demonstrates the bioactive compound of A.cinnampmea and illustrates the mechanism by which it confers its anti-inflammatory activity(Hsieh et al.,2010).Besides the antiinflammation activity study,the hepatoprotective efficacy of antroquinonol and ethanolic extracts of mycelia of A.cinnamomea(EMAC) in vitro and in vivo by us.Based on the results obtained in our study,antroquinonol pretreatment significantly inhibited ethanol-induced AST,ALT,ROS,NO,MDA production and GSH depletion in Hep G2 cells.Western blot and RT-PCR analysis showed that antroquinonol enhanced Nrf-2 activation and its downstream antioxidant gene HO-1 via MAPK pathway.This mechanism was then confirmed in vivo in an acute ethanol intoxicated mouse model:serum ALT and AST production,hepatocellular lipid peroxidation and GSH depletion was prevented by EMAC in a dose-dependent manner.EMAC significantly enhanced HO-1 and Nrf-2 activation via MAPKs consistent with in vitro studies.Ethanol-induced hepatic swelling and hydropic degeneration of hepatocytes was significantly inhibited by EMAC in a dose-dependent manner.These results provide a scientific basis for the hepatoprotective effects of A.cinnamomea.Data also imply that antroquinonol,a potent bioactive compound may be responsible for the hepatoprotective activity of A.cinnamomea.This study highly supported our traditional knowledge that A.cinnamomea as a potential candidate for the treatment of alcoholic liver diseases(Senthil Kumar et al.,2011).On the other hand,an index of 13 representative metabolites from the ethanol extract of A.cinnamomea fruiting body was established for use in quality evaluation.Most of the index compounds selected,particularly the ergostane-type triterpenoids and polyacetylenes,possess good antiinflammation activity.A comparison of the metabolite profiles of different ethanol extracts from A.cinnamomea strains showed similar metabolites when the strains were grown on the original host wood(Cinnamomum kanehirai) and harvested after the same culture time period(9 months).Furthermore,the amounts of typical ergostane-type triterpenoids in A.cinnamomea increased with culture age.Culture substrates also influenced metabolite synthesis;with the same culture age,A.cinnamomea grown on the original host wood produced a richer array of metabolites than A.cinnamomea cultured on other wood species.We conclude that analysis of a fixed group of compounds including triterpenoids,benzolics,and polyacetylenes constitutes a suitable,reliable system to evaluate the quality of ethanol extract from A.cinnamomea fruiting bodies.The evaluation system established in this study may provide a platform for analysisof the products of A.cinnamomea(Lin et al.,2011).Besides fruiting bodies,we selected an A.cinnamomea mycelium health food product,which was produced by solid-state culture and certificate by Taiwan's Department of Health as "National Health Food",as the target for investigation.14 representative metabolites of A.cinnamomea mycelium(EMAC) were selected as index compounds to establish the metabolite profile for evaluation of EMAC product quality.It was also demonstrated that EMAC administration significantly reduced liver inflammation and serum oxidative stress in vivo.4-Acetylantroquinonol B obtained by a bioactivity-guided fractionation from EMAC was able to not only inhibit LPS-induced nitric oxide formation in macrophages but also protect against ethanol-induced oxidative stress in liver cells.The results suggest this A.cinnamomea product might be a potent antioxidative and anti-inflammatory supplement for chemoprevention(Wang et al.,2013).Recently,we found that Antrodin C(ADC),a maleimide derivative isolated from A.cinnamomea health food product inhibits TGF-β1-induced Epithelial-to-mesenchymal transition(EMT) and breast cancer cell metastasis in vitro.Our data suggested that ADC attenuates the TGF-β1-induced EMT,migration and invasion of human breast carcinoma through the suppression of Smad2/3 and β-catenin signaling pathways(Senthil Kumar et al.,2015).The similar activity was also approved for 2,3,5-trimethoxy-4-cresol(TMC),which was isolated from solid-state cultured mycelium of A.cinnamomea(strain no.LEACS-002).We demonstrated that TMC effectively suppresses movement,migration and invasion of lung cancer cells,and achieves an anti-cancer metastasis effect(Lin et al.,2015).
引文
Hsieh Y.-H.,F.-H.Chu,Y.-S.Wang,S.-C.Chien,S.-T.Chang,J.-F.Shaw,C.-Y.Chen,W.-W.Hsiao,Y.-H Kuo*,S.-Y.Wang*.2010.Antrocamphin A,an anti-inflammatory principal from the fruiting body of Taiwanofungus camphoratus,and its Mechanisms.J.Agric.Food Chem.58(5):3153-3158.
Lin C.-C.,K.J.Senthil Kumar,J.-W.Liao,Y.-H.Kuo and S.-Y.Wang*2015.Genotoxic,teratotoxic and oral toxic assessments of Antrodia cinnamomea health food product(Leader Deluxe Antrodia cinnamomea).Toxicolg Reports2(2015):1409-1417.
Lin T.-Y.,C.-Y.Chen,S.-C.Chien,W.-W.Hsiao,F.-H.Chu,W.-H.Li,C.-C.Lin,J.-F.Shaw,S.-Y.Wang*2011.Metabolite profiles for Antrodia cinnamomea fruiting bodies harvested at different culture ages and substrates from different wood.J.Agric.and Food Chem.59:7626-7635.
Lin,C.-C.,C.-C.Chen·Y.-H.Kuo,J.-T Kuo,K.J.Senthil Kumar,S.-Y.Wang*2015.2,3,5-Trimethoxy-4-cresol,an anti-metastatic constituent from the solid-state cultured mycelium of Antrodia cinnamomea and its mechanism.J Nat Med69:513-521.
Senthil Kumar,K.J.,M.,Gokila Vani,P.-J.Chueh,J.-L.Mau,S.-Y.Wang*2015.Antrodin C Inhibits epithelial-to-mesenchymal transition and metastasis of breast cancer cells via suppression of smad2/3 andβ-catenin signaling pathways.PLo S One 10(2):e0117111.
Wang H.-C.,F.-H.Chu,S.-C.Chian,J.-W.Liao,H.-W.Hsieh,W.-H.Li,C.C.Lin,J.-F.Shaw,Y.-H.Kuo,S.Y.Wang*2013.Establishment of the metabolite profile for an Antrodia cinnamomea health food product and investigation of its chemoprevention activity.J.Agric.Food Chem.61(36):8556-8564.