Background:Previously,we showed that total flavonoids from astragalus(TFA)had beneficial effects against transforming growth factor(TGF)-β1-mediated fibrosis,but whether these effects involved autophagy is not known....Background:Previously,we showed that total flavonoids from astragalus(TFA)had beneficial effects against transforming growth factor(TGF)-β1-mediated fibrosis,but whether these effects involved autophagy is not known.We attempt to explore the effects of TFA on autophagy in an animal model of idiopathic pulmonary fibrosis(IPF)induced by bleomycin,and to look for TFA components that may have an effect on autophagy.Methods:C57BL/6 mice were randomized to the sham group(SG),model group,low-dose TFA group(LDTG),and high-dose of TFA group(HDTG).The A549 cell line was treated with the TFA components including formononetin,calycosin,isorhamnetin,kaempferol,and quercetin.Lung tissues and cells were examined by histology,immunohistochemistry(anti-TGF-β1,α-smooth muscle actin(SMA),and cadherin),immunofluorescence(microtubule-associated protein light chain 3(LC3)),hydroxyproline content,and immunoblotting(smad3,smad7,phosphoinositide 3-kinase(PI3K),α-SMA,E-cadherin,beclin-1,and LC3-Ⅱ).Results:In vivo,TFA inhibited TGF-β1 expression and decreased collagen content in lung tissues induced by bleomycin.TFA increased autophagy following suppression of the smad pathway.In vitro,quercetin inhibited the epithelial-mesenchymal transition(EMT)of A549 cells induced by TGF-β1 through suppression of the smad pathway.Autophagy was also increased by quercetin through inhibition of the AKT/mTOR pathway,but without change in PI3K expression.Formononetin,calycosin,isorhamnetin,and kaempferol had no such effects.Conclusion:TFA can alleviate bleomycin-induced PF in C57BL/6 mice via enhanced autophagy.The smad and AKT/mTOR pathways are possibly involved in these effects.Quercetin was the main active compound in TFA.展开更多
To address the significant threat of aflatoxin B1(AFB1)contamination to global food safety,this study innovatively developed a fluorescent aptasensor by synergistically integrating Y-shaped DNA nanostructures with the...To address the significant threat of aflatoxin B1(AFB1)contamination to global food safety,this study innovatively developed a fluorescent aptasensor by synergistically integrating Y-shaped DNA nanostructures with the CRISPR-Cas12a system.The sensor employs an AFB1-specific aptamer(Apt)as a molecular switch,triggering a cascade signal amplification mechanism upon target binding:(1)a strand displacement reactions(SDR)generates Y-shaped DNA assemblies enriched with activation sequence,which precisely activate Cas12a′s transcleavage activity to fragment FAM-labeled signal probes(FAM-sDNA);(2)magnetic beads(MBs)separation enables efficient enrichment of target complexes,while graphene oxide(GO)suppresses background signals via π-π stacking adsorption of un-cleaved probes.Under optimized conditions,the sensor demonstrated a broad linear range(0.05-500 ng/mL,R^(2)=0.995)with an ultralow detection limit of 0.022 ng/mL.Spike-and-recovery tests in complex food matrices(peanuts,corn flour et al.)yielded recoveries of 88.96%-113.55%and relative standard deviations(RSD)below 10%.The t-test confirmed no significant difference between the sensor results and high-performance liquid chromatography(HPLC)measurements.The platform exhibits high sensitivity,specificity(against Aflatoxin G1,Fumonisin B1,Zearalenone,etc.),and reproducibility(RSD=0.69%)and repeatability(RSD=0.59%),offering a robust,high-performance solution the detection of trace mycotoxins in complex food matrices.展开更多
基金the National Natural Science Foundation of China(81760817 and 81260587)the Science and Technology Foundation of Guizhou Province(Qian Science LH[2016]7119).
文摘Background:Previously,we showed that total flavonoids from astragalus(TFA)had beneficial effects against transforming growth factor(TGF)-β1-mediated fibrosis,but whether these effects involved autophagy is not known.We attempt to explore the effects of TFA on autophagy in an animal model of idiopathic pulmonary fibrosis(IPF)induced by bleomycin,and to look for TFA components that may have an effect on autophagy.Methods:C57BL/6 mice were randomized to the sham group(SG),model group,low-dose TFA group(LDTG),and high-dose of TFA group(HDTG).The A549 cell line was treated with the TFA components including formononetin,calycosin,isorhamnetin,kaempferol,and quercetin.Lung tissues and cells were examined by histology,immunohistochemistry(anti-TGF-β1,α-smooth muscle actin(SMA),and cadherin),immunofluorescence(microtubule-associated protein light chain 3(LC3)),hydroxyproline content,and immunoblotting(smad3,smad7,phosphoinositide 3-kinase(PI3K),α-SMA,E-cadherin,beclin-1,and LC3-Ⅱ).Results:In vivo,TFA inhibited TGF-β1 expression and decreased collagen content in lung tissues induced by bleomycin.TFA increased autophagy following suppression of the smad pathway.In vitro,quercetin inhibited the epithelial-mesenchymal transition(EMT)of A549 cells induced by TGF-β1 through suppression of the smad pathway.Autophagy was also increased by quercetin through inhibition of the AKT/mTOR pathway,but without change in PI3K expression.Formononetin,calycosin,isorhamnetin,and kaempferol had no such effects.Conclusion:TFA can alleviate bleomycin-induced PF in C57BL/6 mice via enhanced autophagy.The smad and AKT/mTOR pathways are possibly involved in these effects.Quercetin was the main active compound in TFA.
基金funded by the Key Scientific and Technological Project of Henan Province(242102320270)Special Project for Collaborative Innovation of Zhengzhou(21ZZXTCX15)Henan University of Technology Postdoctoral Scientific Research Fund(21450082).
文摘To address the significant threat of aflatoxin B1(AFB1)contamination to global food safety,this study innovatively developed a fluorescent aptasensor by synergistically integrating Y-shaped DNA nanostructures with the CRISPR-Cas12a system.The sensor employs an AFB1-specific aptamer(Apt)as a molecular switch,triggering a cascade signal amplification mechanism upon target binding:(1)a strand displacement reactions(SDR)generates Y-shaped DNA assemblies enriched with activation sequence,which precisely activate Cas12a′s transcleavage activity to fragment FAM-labeled signal probes(FAM-sDNA);(2)magnetic beads(MBs)separation enables efficient enrichment of target complexes,while graphene oxide(GO)suppresses background signals via π-π stacking adsorption of un-cleaved probes.Under optimized conditions,the sensor demonstrated a broad linear range(0.05-500 ng/mL,R^(2)=0.995)with an ultralow detection limit of 0.022 ng/mL.Spike-and-recovery tests in complex food matrices(peanuts,corn flour et al.)yielded recoveries of 88.96%-113.55%and relative standard deviations(RSD)below 10%.The t-test confirmed no significant difference between the sensor results and high-performance liquid chromatography(HPLC)measurements.The platform exhibits high sensitivity,specificity(against Aflatoxin G1,Fumonisin B1,Zearalenone,etc.),and reproducibility(RSD=0.69%)and repeatability(RSD=0.59%),offering a robust,high-performance solution the detection of trace mycotoxins in complex food matrices.
