OBJECTIVE Glycyrrhetinic acid(GA),one of the main bioactive constituents of the famous Chinese medicinal herb Glycyrrhizauralensis Fisch,presents potent cytotoxicity to various cancer cells both in vitro and in vivo.H...OBJECTIVE Glycyrrhetinic acid(GA),one of the main bioactive constituents of the famous Chinese medicinal herb Glycyrrhizauralensis Fisch,presents potent cytotoxicity to various cancer cells both in vitro and in vivo.Herein,we′d like to determine whether GA triggers autophagy in non-small cell lung cancer cells and the mechanisms involved.METHODS Cell proliferation was determined by MTT assay and colony formation.AnnexinⅤ/PI staining,Hoechst33342 staining,and Western blotting were used to detect GA-induced apoptosis.GA-induced autophagy was measured by expression of the lipid modification of light chain-3(LC3)and transfected with GFP-LC3 or GFP-RFP-LC3 plasmid.Pharmacological regulators,siRNA,and plasmid transfection were used to study the mechanisms of GA-triggered autophagy.RESULTS GA inhibited cell proliferation and induced apoptosis in a concentrationdependent manner in non-small cell lung cancer A549 cells.GA induced autophagyas evidenced by up-regulation of LC3-Ⅱexpression when combined treatment with chloroquine and induction of the red punta after GFP-RFP-LC3 plasmid transfection.Knockdown of autophagy related proteins(ATG)7,ATG 5,or beclin 1by siRNA,the expression of LC3-Ⅱand GFP-LC3 punt atriggered by GA were decreased.Furthermore,the c-Jun N-terminal kinase(JNK)pathways were activated after treatment with GA,and pretreatment with JNK inhibitor SP600125 or silence of JNK pathway by siRNA of JNK or c-jun obviously reduced GA-induced LC3-Ⅱexpression and GFP-LC3 punta formation.GA also stimulate dendoplasmic reticulum stress response by triggering inositol-requiring enzyme 1α(IRE1α)pathway,and knockdown of IRE1αinhibited the activation of JNK pathway and autophagy induced by GA.In addition,GA-induced cell proliferative inhibition and apoptosis were both enhanced when silence of autophagy as well as JNK pathway.CONCLUSION Our study demonstrated,for the first time,that GA induced a cytoprotective autophagy in non-small cell lung cancer cells by activating the IRE1α-JNK pathway,which might decreased the anti-cancer effects of GA.展开更多
OBJECTIVE Rheumatoid arthritis(RA)is the most common inflammatory autoimmune disease,affecting around 1% of the world population.Toll-like receptor 4(TLR4)signalling has been found to be involved in the pathogenesis o...OBJECTIVE Rheumatoid arthritis(RA)is the most common inflammatory autoimmune disease,affecting around 1% of the world population.Toll-like receptor 4(TLR4)signalling has been found to be involved in the pathogenesis of RA.It is a potential therapeutic target for RA treatment.A herbal formula(RL)consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos has traditionally been used in treating various inflammatory disorders.In this study,we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis(CIA)in rats and investigate the involvement of TLR4 signaling in the mode of action of RL in vivo and in vitro.METHODS In vivo anti-arthritic efficacy was evaluated using CIA rats induced by bovine typeⅡ collagen.The treatment groups were treated with various concentrations of RL or positive control indomethacin for 35 d.Clinical signs(hind paw volume and arthritis severity scores),changes in serum inflammatory mediators,histological and radiographic changes of joints were investigated.Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats.The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats,murine RAW264.7macrophages and human THP-1 monocytic cells.RESULTS The severity of arthritis in the CIA rats was significantly attenuated by RL.Histological score and radiographic score were efficiently improved by RL.RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats.RL significantly inhibited the production of various pro-inflammatory mediators,the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines.CONCLUSION RL possesses anti-arthritic effect on collagen-induced arthritis in rats.The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum.The inhibition of the TLR4/TAK1/NF-κB and TLR4/TAK1/MAPK pathways participate in the anti-arthritic effects of RL.This provides a pharmacological justification for the use of RL in the control of various arthritic diseases.Further investigation should be done to develop RL into a modern anti-arthritic agent.展开更多
Spectators who have watched Harry Potter movie series must be fascinated by Hogwarts Castle- the assembly hall hanging hundreds of chandeliers and countless steps of the spiral staircases, exaggerated by animation tec...Spectators who have watched Harry Potter movie series must be fascinated by Hogwarts Castle- the assembly hall hanging hundreds of chandeliers and countless steps of the spiral staircases, exaggerated by animation technologies, show viewers a horrible and fabulous world of wizardry.展开更多
基金The project supported by Science and Technology Development Fund,Macao S.A.R(FDCT)(074/2012/A)the Research Fund of University of Macao(MRG013/WYT/2013/ICMS,MYRG2015-00101-ICMS-QRCMand CPG2014-00012-ICMS)
文摘OBJECTIVE Glycyrrhetinic acid(GA),one of the main bioactive constituents of the famous Chinese medicinal herb Glycyrrhizauralensis Fisch,presents potent cytotoxicity to various cancer cells both in vitro and in vivo.Herein,we′d like to determine whether GA triggers autophagy in non-small cell lung cancer cells and the mechanisms involved.METHODS Cell proliferation was determined by MTT assay and colony formation.AnnexinⅤ/PI staining,Hoechst33342 staining,and Western blotting were used to detect GA-induced apoptosis.GA-induced autophagy was measured by expression of the lipid modification of light chain-3(LC3)and transfected with GFP-LC3 or GFP-RFP-LC3 plasmid.Pharmacological regulators,siRNA,and plasmid transfection were used to study the mechanisms of GA-triggered autophagy.RESULTS GA inhibited cell proliferation and induced apoptosis in a concentrationdependent manner in non-small cell lung cancer A549 cells.GA induced autophagyas evidenced by up-regulation of LC3-Ⅱexpression when combined treatment with chloroquine and induction of the red punta after GFP-RFP-LC3 plasmid transfection.Knockdown of autophagy related proteins(ATG)7,ATG 5,or beclin 1by siRNA,the expression of LC3-Ⅱand GFP-LC3 punt atriggered by GA were decreased.Furthermore,the c-Jun N-terminal kinase(JNK)pathways were activated after treatment with GA,and pretreatment with JNK inhibitor SP600125 or silence of JNK pathway by siRNA of JNK or c-jun obviously reduced GA-induced LC3-Ⅱexpression and GFP-LC3 punta formation.GA also stimulate dendoplasmic reticulum stress response by triggering inositol-requiring enzyme 1α(IRE1α)pathway,and knockdown of IRE1αinhibited the activation of JNK pathway and autophagy induced by GA.In addition,GA-induced cell proliferative inhibition and apoptosis were both enhanced when silence of autophagy as well as JNK pathway.CONCLUSION Our study demonstrated,for the first time,that GA induced a cytoprotective autophagy in non-small cell lung cancer cells by activating the IRE1α-JNK pathway,which might decreased the anti-cancer effects of GA.
基金The project supported by Science,Technology and Innovation Commission of Shenzhen(JCYJ20120829154222473and JCYJ20140807091945050)Research Grants Council of Hong Kong(HKBU 262512)+1 种基金Food and Health Bureau of Hong Kong(HMRF 11122521)Hong Kong Baptist University(FRG2/14-15/056,FRG1/14-15/061)
文摘OBJECTIVE Rheumatoid arthritis(RA)is the most common inflammatory autoimmune disease,affecting around 1% of the world population.Toll-like receptor 4(TLR4)signalling has been found to be involved in the pathogenesis of RA.It is a potential therapeutic target for RA treatment.A herbal formula(RL)consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos has traditionally been used in treating various inflammatory disorders.In this study,we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis(CIA)in rats and investigate the involvement of TLR4 signaling in the mode of action of RL in vivo and in vitro.METHODS In vivo anti-arthritic efficacy was evaluated using CIA rats induced by bovine typeⅡ collagen.The treatment groups were treated with various concentrations of RL or positive control indomethacin for 35 d.Clinical signs(hind paw volume and arthritis severity scores),changes in serum inflammatory mediators,histological and radiographic changes of joints were investigated.Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats.The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats,murine RAW264.7macrophages and human THP-1 monocytic cells.RESULTS The severity of arthritis in the CIA rats was significantly attenuated by RL.Histological score and radiographic score were efficiently improved by RL.RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats.RL significantly inhibited the production of various pro-inflammatory mediators,the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines.CONCLUSION RL possesses anti-arthritic effect on collagen-induced arthritis in rats.The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum.The inhibition of the TLR4/TAK1/NF-κB and TLR4/TAK1/MAPK pathways participate in the anti-arthritic effects of RL.This provides a pharmacological justification for the use of RL in the control of various arthritic diseases.Further investigation should be done to develop RL into a modern anti-arthritic agent.
文摘Spectators who have watched Harry Potter movie series must be fascinated by Hogwarts Castle- the assembly hall hanging hundreds of chandeliers and countless steps of the spiral staircases, exaggerated by animation technologies, show viewers a horrible and fabulous world of wizardry.
