OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment t...OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment to construct a myocardial ischemiareperfusion model,and were divided into normal,model,low(10μmol·L^(-1)),medium(20μmol·L^(-1))and high(40μmol·L^(-1))ICA group,and high ICA+inhibitor group(40μmol·L^(-1)+20 nmol·L^(-1)).CCK-8 assay was used to assess the protective ability of ICA against CMEC,and cell migration assay and tube-formation assay were used to detect the migration and generation ability of CMEC.The TCMSP database,Swiss-Target database and literature mining methods were used to col⁃lect ICA-related targets,the GeneCards data⁃base was used to collect target genes related to myocardial ischemia/reperfusion,and Cytoscape 3.8.0 software was used to construct a"drug-tar⁃get-disease"network.The potential targets were imported into STRING 11.5 database to obtain the PPI network.GO and KEGG enrichment analyses were performed on the potential targets using the DAVID database.Molecular docking was performed using AutoDock-vina 1.1.2 soft⁃ware.Western blot detected the expression of related proteins.RESULTS After CMEC was subjected to OGD/R treatment,ICA had a protec⁃tive effect at 10^(-1)60μmol·L^(-1);the results of the cell migration assay showed that each group of ICA could promote the migratory effect of CMEC(P<0.01,P<0.01);and the results of tube-for⁃mation assay showed that each group of ICA could significantly promote the generation of branches(P<0.01)and the capillary length exten⁃sion(P<0.05).Network pharmacology collected a total of 23 ICA action targets,1500 disease tar⁃gets and 12 key targets.GO function enrichment analysis found 85 results.KEGG pathway enrich⁃ment analysis found 53 results,involving AGERAGE signaling pathway,sphingolipid signaling pathway and VEGF signaling pathway.Molecu⁃lar docking results showed that ICA had better binding with core targets PRKCB,PRKCA and PTGS2.Western blot results showed that ICA could regulate the expression of PRKCB,PRKCA and PTGS2 proteins.The results of cell migra⁃tion assay,tube-formation assay and protein expression were reversed after addition of PKC inhibitor.CONCLUSION The potential mecha⁃nism of action of ICA against myocardial isch⁃emia-reperfusion injury may be related to the reg⁃ulation of processes such as CMEC migration and angiogenesis,and it functions through the key target gene PKC.展开更多
目的探讨钆塞酸二钠(Gd-EOB-DTPA)增强磁共振成像(MRI)联合血清甲胎蛋白(AFP)和脱-γ-羧基凝血酶原(PIVKA-Ⅱ)诊断肝细胞癌(HCC)的临床价值。方法2015年12月~2017年9月收治的82例肝病患者,经组织病理学诊断HCC组56例,非HCC组26例。罗氏c...目的探讨钆塞酸二钠(Gd-EOB-DTPA)增强磁共振成像(MRI)联合血清甲胎蛋白(AFP)和脱-γ-羧基凝血酶原(PIVKA-Ⅱ)诊断肝细胞癌(HCC)的临床价值。方法2015年12月~2017年9月收治的82例肝病患者,经组织病理学诊断HCC组56例,非HCC组26例。罗氏cobasR e 601型全自动电化学发光免疫分析系统检测血清AFP,采用LUMIPULSE G1200全自动免疫分析仪酶化学发光法检测血清PIVKA-Ⅱ。全部患者接受MRI检查。采用受试者工作特征曲线(ROC)下面积(AUC)判断各检查诊断的灵敏度、特异度和正确率。结果HCC患者血清AFP和PIVKA-Ⅱ水平分别为34.5(4.5,594.9)ng/ml和63.5(25.0,2082.0)Mau/ml,显著高于非HCC组【分别为3.4(2.2,11.6)ng/ml和23.0(18.8,28.0)Mau/ml,P<0.01】;血清AFP、PIVKA-Ⅱ和Gd-EOB-DTPA增强MRI检查单独诊断HCC的ROC曲线下面积分别为0.763、0.815和0.907;在单项诊断HCC时,血清AFP和PIVKA-Ⅱ的最佳临床诊断截断点分别为14.4ng/ml和40.5 Mau/ml,其诊断HCC的灵敏度、特异度和准确率分别为64.3%、84.6%、67.4%和62.5%、100.0%、74.4%,Gd-EOB-DTPA增强MRI单独诊断HCC的灵敏度、特异度和准确率分别为92.9%、88.5%、91.5%,以Gd-EOB-DTPA增强MRI诊断的效能最高;采用联合试验Ⅰ诊断HCC,其灵敏度为98.2%,特异度为61.5%,准确率为86.6%,采用联合试验Ⅱ诊断HCC的灵敏度为50.0%,特异度为100.0%,准确率为50.0%。联合试验提高了诊断的特异度,但降低了灵敏度。结论应用血清AFP和PIVKA-Ⅱ检测联合Gd-EOB-DTPA增强MRI检查可提高诊断HCC的效能,综合应用三者联合诊断可以提高诊断HCC的正确性。展开更多
Objective:To explore the protective effect of Huoxin Pill(HXP)on acute myocardial ischemia-reperfusion(MIRI)injury in rats.Methods:Seventy-five adult SD rats were divided into the sham-operated group,model group,posit...Objective:To explore the protective effect of Huoxin Pill(HXP)on acute myocardial ischemia-reperfusion(MIRI)injury in rats.Methods:Seventy-five adult SD rats were divided into the sham-operated group,model group,positive drug group(diltiazem hydrochloride,DH),high dose group(24 mg/kg,HXP-H)and low dose group(12 mg/kg,HXP-L)of Huoxin Pill(n=15 for every group)according to the complete randomization method.After 1 week of intragastric administration,the left anterior descending coronary artery of the rat's heart was ligated for 45 min and reperfused for 3 h.Serum was separated and the levels of creatine kinase(CK),creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH),superoxide dismutase(SOD),and malondialdehyde(MDA),hypersensitive C-reactive protein(hs-CRP)and interleukin-1β(IL-1β)were measured.Myocardial ischemia rate,myocardial infarction rate and myocardial no-reflow rate were determined by staining with Evans blue and 2,3,5-triphenyltetrazolium chloride(TTC).Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN)databases were used to screen for possible active compounds of HXP and their potential therapeutic targets;the results of anti-inflammatory genes associated with MIRI were obtained from GeneC ards,Drugbank,Online Mendelian Inheritance in Man(OMIM),and Therapeutic Target Datebase(TTD)databases was performed;Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment were used to analyze the intersected targets;molecular docking was performed using AutoD ock Tools.Western blot was used to detect the protein expression of Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NFκB)/NOD-like receptor protein 3(NLRP3).Results:Compared with the model group,all doses of HXP significantly reduced the levels of LDH,CK and CK-MB(P<0.05,P<0.01);HXP significantly increased serum activity of SOD(P<0.05,P<0.01);all doses of HXP significantly reduced the levels of hs-CRP and IL-1β(P<0.05,P<0.01)and the myocardial infarction rate and myocardial no-reflow rate(P<0.01).GO enrichment analysis mainly involved positive regulation of gene expression,extracellular space and identical protein binding,KEGG pathway enrichment mainly involved PI3K-Akt signaling pathway and lipid and atherosclerosis.Molecular docking results showed that kaempferol and luteolin had a better affinity with TLR4,NFκB and NLRP3 molecules.The protein expressions of TLR4,NFκB and NLRP3 were reduced in the HXP group(P<0.01).Conclusions:HXP has a significant protective effect on myocardial ischemia-reperfusion injury in rats,and its effect may be related to the inhibition of redox response and reduction of the inflammatory response by inhibiting the TLR4/NFκB/NLRP3 signaling pathway.展开更多
基金National Natural Science Foundation of China(82030124)National Natural Science Foundation of China(82174015)Science and Technology Innovation Project of China Academy of Traditional Chinese Medicine(CI2021A04609)。
文摘OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment to construct a myocardial ischemiareperfusion model,and were divided into normal,model,low(10μmol·L^(-1)),medium(20μmol·L^(-1))and high(40μmol·L^(-1))ICA group,and high ICA+inhibitor group(40μmol·L^(-1)+20 nmol·L^(-1)).CCK-8 assay was used to assess the protective ability of ICA against CMEC,and cell migration assay and tube-formation assay were used to detect the migration and generation ability of CMEC.The TCMSP database,Swiss-Target database and literature mining methods were used to col⁃lect ICA-related targets,the GeneCards data⁃base was used to collect target genes related to myocardial ischemia/reperfusion,and Cytoscape 3.8.0 software was used to construct a"drug-tar⁃get-disease"network.The potential targets were imported into STRING 11.5 database to obtain the PPI network.GO and KEGG enrichment analyses were performed on the potential targets using the DAVID database.Molecular docking was performed using AutoDock-vina 1.1.2 soft⁃ware.Western blot detected the expression of related proteins.RESULTS After CMEC was subjected to OGD/R treatment,ICA had a protec⁃tive effect at 10^(-1)60μmol·L^(-1);the results of the cell migration assay showed that each group of ICA could promote the migratory effect of CMEC(P<0.01,P<0.01);and the results of tube-for⁃mation assay showed that each group of ICA could significantly promote the generation of branches(P<0.01)and the capillary length exten⁃sion(P<0.05).Network pharmacology collected a total of 23 ICA action targets,1500 disease tar⁃gets and 12 key targets.GO function enrichment analysis found 85 results.KEGG pathway enrich⁃ment analysis found 53 results,involving AGERAGE signaling pathway,sphingolipid signaling pathway and VEGF signaling pathway.Molecu⁃lar docking results showed that ICA had better binding with core targets PRKCB,PRKCA and PTGS2.Western blot results showed that ICA could regulate the expression of PRKCB,PRKCA and PTGS2 proteins.The results of cell migra⁃tion assay,tube-formation assay and protein expression were reversed after addition of PKC inhibitor.CONCLUSION The potential mecha⁃nism of action of ICA against myocardial isch⁃emia-reperfusion injury may be related to the reg⁃ulation of processes such as CMEC migration and angiogenesis,and it functions through the key target gene PKC.
文摘目的探讨钆塞酸二钠(Gd-EOB-DTPA)增强磁共振成像(MRI)联合血清甲胎蛋白(AFP)和脱-γ-羧基凝血酶原(PIVKA-Ⅱ)诊断肝细胞癌(HCC)的临床价值。方法2015年12月~2017年9月收治的82例肝病患者,经组织病理学诊断HCC组56例,非HCC组26例。罗氏cobasR e 601型全自动电化学发光免疫分析系统检测血清AFP,采用LUMIPULSE G1200全自动免疫分析仪酶化学发光法检测血清PIVKA-Ⅱ。全部患者接受MRI检查。采用受试者工作特征曲线(ROC)下面积(AUC)判断各检查诊断的灵敏度、特异度和正确率。结果HCC患者血清AFP和PIVKA-Ⅱ水平分别为34.5(4.5,594.9)ng/ml和63.5(25.0,2082.0)Mau/ml,显著高于非HCC组【分别为3.4(2.2,11.6)ng/ml和23.0(18.8,28.0)Mau/ml,P<0.01】;血清AFP、PIVKA-Ⅱ和Gd-EOB-DTPA增强MRI检查单独诊断HCC的ROC曲线下面积分别为0.763、0.815和0.907;在单项诊断HCC时,血清AFP和PIVKA-Ⅱ的最佳临床诊断截断点分别为14.4ng/ml和40.5 Mau/ml,其诊断HCC的灵敏度、特异度和准确率分别为64.3%、84.6%、67.4%和62.5%、100.0%、74.4%,Gd-EOB-DTPA增强MRI单独诊断HCC的灵敏度、特异度和准确率分别为92.9%、88.5%、91.5%,以Gd-EOB-DTPA增强MRI诊断的效能最高;采用联合试验Ⅰ诊断HCC,其灵敏度为98.2%,特异度为61.5%,准确率为86.6%,采用联合试验Ⅱ诊断HCC的灵敏度为50.0%,特异度为100.0%,准确率为50.0%。联合试验提高了诊断的特异度,但降低了灵敏度。结论应用血清AFP和PIVKA-Ⅱ检测联合Gd-EOB-DTPA增强MRI检查可提高诊断HCC的效能,综合应用三者联合诊断可以提高诊断HCC的正确性。
基金Supported by the National Natural Science Foundation of China (No.82174015 and No.82030124)Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences (No.CI2021A04609)。
文摘Objective:To explore the protective effect of Huoxin Pill(HXP)on acute myocardial ischemia-reperfusion(MIRI)injury in rats.Methods:Seventy-five adult SD rats were divided into the sham-operated group,model group,positive drug group(diltiazem hydrochloride,DH),high dose group(24 mg/kg,HXP-H)and low dose group(12 mg/kg,HXP-L)of Huoxin Pill(n=15 for every group)according to the complete randomization method.After 1 week of intragastric administration,the left anterior descending coronary artery of the rat's heart was ligated for 45 min and reperfused for 3 h.Serum was separated and the levels of creatine kinase(CK),creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH),superoxide dismutase(SOD),and malondialdehyde(MDA),hypersensitive C-reactive protein(hs-CRP)and interleukin-1β(IL-1β)were measured.Myocardial ischemia rate,myocardial infarction rate and myocardial no-reflow rate were determined by staining with Evans blue and 2,3,5-triphenyltetrazolium chloride(TTC).Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN)databases were used to screen for possible active compounds of HXP and their potential therapeutic targets;the results of anti-inflammatory genes associated with MIRI were obtained from GeneC ards,Drugbank,Online Mendelian Inheritance in Man(OMIM),and Therapeutic Target Datebase(TTD)databases was performed;Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment were used to analyze the intersected targets;molecular docking was performed using AutoD ock Tools.Western blot was used to detect the protein expression of Toll-like receptor 4(TLR4)/nuclear factor kappa-B(NFκB)/NOD-like receptor protein 3(NLRP3).Results:Compared with the model group,all doses of HXP significantly reduced the levels of LDH,CK and CK-MB(P<0.05,P<0.01);HXP significantly increased serum activity of SOD(P<0.05,P<0.01);all doses of HXP significantly reduced the levels of hs-CRP and IL-1β(P<0.05,P<0.01)and the myocardial infarction rate and myocardial no-reflow rate(P<0.01).GO enrichment analysis mainly involved positive regulation of gene expression,extracellular space and identical protein binding,KEGG pathway enrichment mainly involved PI3K-Akt signaling pathway and lipid and atherosclerosis.Molecular docking results showed that kaempferol and luteolin had a better affinity with TLR4,NFκB and NLRP3 molecules.The protein expressions of TLR4,NFκB and NLRP3 were reduced in the HXP group(P<0.01).Conclusions:HXP has a significant protective effect on myocardial ischemia-reperfusion injury in rats,and its effect may be related to the inhibition of redox response and reduction of the inflammatory response by inhibiting the TLR4/NFκB/NLRP3 signaling pathway.
