The composition of serum is extremely complex,which complicates the discovery of new pharmacodynamic biomarkers via serum proteome for disease prediction and diagnosis.Recently,nanoparticles have been reported to effi...The composition of serum is extremely complex,which complicates the discovery of new pharmacodynamic biomarkers via serum proteome for disease prediction and diagnosis.Recently,nanoparticles have been reported to efficiently reduce the proportion of high-abundance proteins and enrich lowabundance proteins in serum.Here,we synthesized a silica-coated iron oxide nanoparticle and developed a highly efficient and reproducible protein corona(PC)-based proteomic analysis strategy to improve the range of serum proteomic analysis.We identified 1,070 proteins with a median coefficient of variation of 12.56%using PC-based proteomic analysis,which was twice the number of proteins identified by direct digestion.There were also more biological processes enriched with these proteins.We applied this strategy to identify more pharmacodynamic biomarkers on collagen-induced arthritis(CIA)rat model treated with methotrexate(MTX).The bioinformatic results indicated that 485 differentially expressed proteins(DEPs)were found in CIA rats,of which 323 DEPs recovered to near normal levels after treatment with MTX.This strategy can not only help enhance our understanding of the mechanisms of disease and drug action through serum proteomics studies,but also provide more pharmacodynamic biomarkers for disease prediction,diagnosis,and treatment.展开更多
Natural products from medicinal plants serve as an invaluable resource for drug discovery and development.However,low-abundance natural products are often understudied due to the challenges of obtaining sufficient qua...Natural products from medicinal plants serve as an invaluable resource for drug discovery and development.However,low-abundance natural products are often understudied due to the challenges of obtaining sufficient quantities for pharmacological testing in cells or animals.Additionally,their complex stereochemistry and functional groups make chemical synthesis and purification difficult.In this study,we showcased the power of biosynthetic approaches to explore these underexplored compounds,using the low-abundance polyphyllin trillin 6′-O-glucoside from Paris polyphylla as an example.We identified two trillin 6′-O-glucosyltransferases required for its biosynthesis and successfully reconstructed the entire pathway in Nicotiana benthamiana.We demonstrated that trillin 6′-O-glucoside exhibits anti-bacterial activity comparable to major polyphyllins like polyphyllinsⅠ,Ⅱ,and Ⅶ.Notably,it also showed much lower hemolytic activity,a common side effect of those major polyphyllins.Together,our study underscores the advantages of employing biosynthetic approaches to explore natural products that exist in low or trace abundances yet possess equally important pharmacological activities.展开更多
Background:Acetaminophen(APAP)-induced hepatotoxicity has attracted considerable attention in clinical settings due to the limited treatment options available.Liensinine stands out as a key alkaloid known for its phar...Background:Acetaminophen(APAP)-induced hepatotoxicity has attracted considerable attention in clinical settings due to the limited treatment options available.Liensinine stands out as a key alkaloid known for its pharmaceutical activities.However,the role of liensinine in mitigating APAP-induced liver injury remains unclear.Objective:The aim of the study was to explore the protective effects of liensinine against APAP-induced liver injury.Methods:C57BL/6 male mice were treated with a dose of 200 mg/kg N-acetylcysteine or varying doses of liensinine(10 or 20 mg/kg)for seven consecutive days.APAP(400 mg/kg,i.g.)was then administered to induce liver damage for 12 hours.Blood samples and hepatic tissues were collected for further analysis.Liver enzyme levels and histopathological analysis were employed to assess liver injury.RNA-seq was conducted to evaluate the dynamic changes in gene expression.Biochemical assays were used to measure oxidative stress and inflammation,while the TUNEL assay was performed to assess hepatocyte apoptosis.Results:The results demonstrated that the administration of liensinine mitigated serum liver enzyme levels and tissue damage resulting from APAP overdose.Transcriptome analysis revealed significant and coordinated changes in genes related to the peroxisome proliferator-activated receptor signaling pathway,mitogen-activated protein kinase signaling pathway,and apoptosis pathway in response to APAP-induced hepatotoxicity.The expression alterations of key genes within these three pathways,associated with inflammation,oxidative stress,and cell apoptosis,were reversed by liensinine,indicating its potential in alleviating APAP-induced liver damage through multiple signaling pathways.This suggests the diverse therapeutic effects of liensinine,including inflammation suppression,oxidative stress reduction,and cell apoptosis inhibition.Indeed,pretreatment with liensinine effectively reduced inflammatory cytokines,oxidative stress indicators,and apoptotic cells induced by APAP.Conclusions:Liensinine mitigates APAP-induced hepatotoxicity in mice through multifaceted pathways,providing anti-inflammatory,antioxidant,and anti-apoptotic benefits.展开更多
基金support from the National Key Research and Development Program of China(Grant Nos.:2020YFA0908000 and 2020YFE0205100)the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(Grant No.:ZYYCXTD-C-202002)+3 种基金the National Natural Science Foundation of China(Grant Nos.:82074098,82173914,and 82141001)the CACMS Innovation Fund(Grant Nos.:CI2021A05101 and CI2021A05104)the Fundamental Research Funds for the Central Public Welfare Research Institutes(Grant Nos.:ZZ15-YQ-065,ZZ14-YQ-058,ZZ14-YQ-050,ZZ14-YQ-051,ZZ14-YQ-052,ZZ14-ND-010,ZZ15-ND-10,and ZZ14-FL-002)the Chinese Academy of Sciences(Grant No.:YJKYYQ20210025).
文摘The composition of serum is extremely complex,which complicates the discovery of new pharmacodynamic biomarkers via serum proteome for disease prediction and diagnosis.Recently,nanoparticles have been reported to efficiently reduce the proportion of high-abundance proteins and enrich lowabundance proteins in serum.Here,we synthesized a silica-coated iron oxide nanoparticle and developed a highly efficient and reproducible protein corona(PC)-based proteomic analysis strategy to improve the range of serum proteomic analysis.We identified 1,070 proteins with a median coefficient of variation of 12.56%using PC-based proteomic analysis,which was twice the number of proteins identified by direct digestion.There were also more biological processes enriched with these proteins.We applied this strategy to identify more pharmacodynamic biomarkers on collagen-induced arthritis(CIA)rat model treated with methotrexate(MTX).The bioinformatic results indicated that 485 differentially expressed proteins(DEPs)were found in CIA rats,of which 323 DEPs recovered to near normal levels after treatment with MTX.This strategy can not only help enhance our understanding of the mechanisms of disease and drug action through serum proteomics studies,but also provide more pharmacodynamic biomarkers for disease prediction,diagnosis,and treatment.
基金supported by the National Key Research and Development Program of China(2020YFA0908000)CAMS Innovation Fund(CI2023E002,CI2021A05101,CI2021B014,CI2021A05104 and CI2023E005TS06)+1 种基金Scientific and technological innovation project of China Academy of Chinese Medical Sciences(CI2023D008)Fundamental Research Funds for the Central Public Welfare Research Institutes(ZZ14-YQ-051,ZZ14-YQ-052,ZZ17-YQ-047,ZZ15-ND-10,ZZ16-ND-10-12,ZZ17-ND-10-02,ZZ17-ND-10-16 and ZZ16-XRZ-103)。
文摘Natural products from medicinal plants serve as an invaluable resource for drug discovery and development.However,low-abundance natural products are often understudied due to the challenges of obtaining sufficient quantities for pharmacological testing in cells or animals.Additionally,their complex stereochemistry and functional groups make chemical synthesis and purification difficult.In this study,we showcased the power of biosynthetic approaches to explore these underexplored compounds,using the low-abundance polyphyllin trillin 6′-O-glucoside from Paris polyphylla as an example.We identified two trillin 6′-O-glucosyltransferases required for its biosynthesis and successfully reconstructed the entire pathway in Nicotiana benthamiana.We demonstrated that trillin 6′-O-glucoside exhibits anti-bacterial activity comparable to major polyphyllins like polyphyllinsⅠ,Ⅱ,and Ⅶ.Notably,it also showed much lower hemolytic activity,a common side effect of those major polyphyllins.Together,our study underscores the advantages of employing biosynthetic approaches to explore natural products that exist in low or trace abundances yet possess equally important pharmacological activities.
基金supported by National Natural Science Foundation of China(No.82173914,32300282)Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(CI2023E002)Fundamental Research Funds for the Central Public Welfare Research Institutes(ZZ14-YQ-051,ZZ14-YQ-052,ZZ16-ND-10-12).
文摘Background:Acetaminophen(APAP)-induced hepatotoxicity has attracted considerable attention in clinical settings due to the limited treatment options available.Liensinine stands out as a key alkaloid known for its pharmaceutical activities.However,the role of liensinine in mitigating APAP-induced liver injury remains unclear.Objective:The aim of the study was to explore the protective effects of liensinine against APAP-induced liver injury.Methods:C57BL/6 male mice were treated with a dose of 200 mg/kg N-acetylcysteine or varying doses of liensinine(10 or 20 mg/kg)for seven consecutive days.APAP(400 mg/kg,i.g.)was then administered to induce liver damage for 12 hours.Blood samples and hepatic tissues were collected for further analysis.Liver enzyme levels and histopathological analysis were employed to assess liver injury.RNA-seq was conducted to evaluate the dynamic changes in gene expression.Biochemical assays were used to measure oxidative stress and inflammation,while the TUNEL assay was performed to assess hepatocyte apoptosis.Results:The results demonstrated that the administration of liensinine mitigated serum liver enzyme levels and tissue damage resulting from APAP overdose.Transcriptome analysis revealed significant and coordinated changes in genes related to the peroxisome proliferator-activated receptor signaling pathway,mitogen-activated protein kinase signaling pathway,and apoptosis pathway in response to APAP-induced hepatotoxicity.The expression alterations of key genes within these three pathways,associated with inflammation,oxidative stress,and cell apoptosis,were reversed by liensinine,indicating its potential in alleviating APAP-induced liver damage through multiple signaling pathways.This suggests the diverse therapeutic effects of liensinine,including inflammation suppression,oxidative stress reduction,and cell apoptosis inhibition.Indeed,pretreatment with liensinine effectively reduced inflammatory cytokines,oxidative stress indicators,and apoptotic cells induced by APAP.Conclusions:Liensinine mitigates APAP-induced hepatotoxicity in mice through multifaceted pathways,providing anti-inflammatory,antioxidant,and anti-apoptotic benefits.
