OBJECTIVE To identify the bioactive anti-angiogenic constitutes targeting tumor endothelial cells(TECs)in Shenmai Injection(SMI).METHEODS For pharmacokinetic(PK)studies,Balb/c mice harboring human colorectal cancer(Lo...OBJECTIVE To identify the bioactive anti-angiogenic constitutes targeting tumor endothelial cells(TECs)in Shenmai Injection(SMI).METHEODS For pharmacokinetic(PK)studies,Balb/c mice harboring human colorectal cancer(LoVo)xenografts were treated with SMI 10 mL·kg^-1 daily for 1 or 8 d.Multidimensional PK profiles of ginsenosides in plasma,subcutaneous tumors,and TECs were investigated.For PD studies,the tumor-bearing mice Intravital multi-photon imaging and CD31 immunofluorescence staining were used to evaluate the number of microves⁃sels and braches.Double staining of CD31 and α-SMA was performed to evaluate pericytes coverage ratios around vessels.ELISA was performed to determine the concentrations of VEGF and FGF in tumor tissues.For synergistic anti-tumor study,the tumor-bearing mice were treated with SMI 10 mL·kg^-1 daily,Rd 5 mg·kg^-1 daily with or without 5-FU 15 mg·kg^-1 every 3 d for 20 d.HPLC-MS/MS was used to determine the concentrations of 5-FU in plasma and tumor tissues.RESULTS SMI decreased the number of microvessels(P<0.05)and vessel branches(P<0.05)and improved vascular pericytes coverage(P<0.05).PK studies showed that the concentrations of protopanaxadiol-type(PPD)ginsenosides(Rb1,Rb2/Rb3,Rc,and Rd)in both,plasma and tumors,were higher than those of protopanaxatriol-type(Rg1 and Re)and oleanane-type(Ro)ginsenosides.Among PPD ginsenosides,Rd exhibited the greatest concentrations in tumors and TECs after repeated injection.In fact,the proportion of Rd in the detectable components of SMI gradually increased in the following order:SMI formula(2.8%),plasma(16.0%),tumor tissues(34.3%),and TECs(40.3%).In vivo bioactivity results showed that Rd 5 mg·kg^-1 daily significantly decreased the number of microvessels(P<0.05)and vessel branches(P<0.05)and increased pericytes coverage(P<0.05)while Rd 0.5 mg·kg^-1 daily,Rb1 and Rg1 had no significant effect on them.Rd 5 mg·kg^-1 suppressed the expression of VEGF and FGF simultaneously.Rd 5 mg·kg^-1 enhanced the antitumor effect of 5-FU via increasing the distribution of 5-FU in tumor tissues(P<0.05)in xenograft mice.CONCLUSION Ginsenoside Rd may be the major bioactive anti-angiogenic constituent targeting TECs after SMI treatment.展开更多
OBJECTIVE Shenmai Injection(SMI)is widely used in the treatment of cardiovascular diseases,such as heart failure and myocardial ischemia.In clinic,SMI showed protective effects on doxorubicin(Dox)-induced cardiac toxi...OBJECTIVE Shenmai Injection(SMI)is widely used in the treatment of cardiovascular diseases,such as heart failure and myocardial ischemia.In clinic,SMI showed protective effects on doxorubicin(Dox)-induced cardiac toxicity.In current study,we investigate the mitochondrial protective mechanisms of SMI on Dox-induced myocardial injury.METHODS C57BL/6 mice were divided into four groups:①control group;②Dox injury group;③SMI+Dox group and dexrazoxane(DRZ)+Dox group.Dex was a positive control.Myocardial injury was evaluated by echocardiography,HE and TUNEL staining,myocardial markers measurement.H9C2 cardiomyocytes pretreatment with SMI for 24 h were exposed to Dox.Cell viability and apoptosis were measured by CCK8,Hoechst33342 staining,and Annexin V/PI staining.MitoSOX,mitochondrial membrane potential,and mitochondrial respiratory function were measured to evaluate mito⁃chondrial function.RESULTS SMI decreased mortality rate of Dox-injected mice,serum CK and CK-MB levels in vivo.SMI significantly prevented Dox-induced cardiac dysfunction and apoptosis and increased expression level of PI3K,p-Akt,and p-GSK-3β.Moreover,SMI significantly inhibited Dox-induced apoptosis,mitochondrial ROS production,and reduction of mitochondrial membrane potential in H9C2 cells.Mechanismly,the cardio-protective effect of SMI was suppressed by PI3K inhibitor LY294002.CONCLUSION SMI prevents Dox-induced cardiotoxicity and mitochondrial damage through activation of PI3K/Akt signaling pathway.展开更多
基金National Nature Science Foundation of China(81773989and 81530098)
文摘OBJECTIVE To identify the bioactive anti-angiogenic constitutes targeting tumor endothelial cells(TECs)in Shenmai Injection(SMI).METHEODS For pharmacokinetic(PK)studies,Balb/c mice harboring human colorectal cancer(LoVo)xenografts were treated with SMI 10 mL·kg^-1 daily for 1 or 8 d.Multidimensional PK profiles of ginsenosides in plasma,subcutaneous tumors,and TECs were investigated.For PD studies,the tumor-bearing mice Intravital multi-photon imaging and CD31 immunofluorescence staining were used to evaluate the number of microves⁃sels and braches.Double staining of CD31 and α-SMA was performed to evaluate pericytes coverage ratios around vessels.ELISA was performed to determine the concentrations of VEGF and FGF in tumor tissues.For synergistic anti-tumor study,the tumor-bearing mice were treated with SMI 10 mL·kg^-1 daily,Rd 5 mg·kg^-1 daily with or without 5-FU 15 mg·kg^-1 every 3 d for 20 d.HPLC-MS/MS was used to determine the concentrations of 5-FU in plasma and tumor tissues.RESULTS SMI decreased the number of microvessels(P<0.05)and vessel branches(P<0.05)and improved vascular pericytes coverage(P<0.05).PK studies showed that the concentrations of protopanaxadiol-type(PPD)ginsenosides(Rb1,Rb2/Rb3,Rc,and Rd)in both,plasma and tumors,were higher than those of protopanaxatriol-type(Rg1 and Re)and oleanane-type(Ro)ginsenosides.Among PPD ginsenosides,Rd exhibited the greatest concentrations in tumors and TECs after repeated injection.In fact,the proportion of Rd in the detectable components of SMI gradually increased in the following order:SMI formula(2.8%),plasma(16.0%),tumor tissues(34.3%),and TECs(40.3%).In vivo bioactivity results showed that Rd 5 mg·kg^-1 daily significantly decreased the number of microvessels(P<0.05)and vessel branches(P<0.05)and increased pericytes coverage(P<0.05)while Rd 0.5 mg·kg^-1 daily,Rb1 and Rg1 had no significant effect on them.Rd 5 mg·kg^-1 suppressed the expression of VEGF and FGF simultaneously.Rd 5 mg·kg^-1 enhanced the antitumor effect of 5-FU via increasing the distribution of 5-FU in tumor tissues(P<0.05)in xenograft mice.CONCLUSION Ginsenoside Rd may be the major bioactive anti-angiogenic constituent targeting TECs after SMI treatment.
基金National Natural Science Foundation of China(8177401781202779+2 种基金81973624)Natural Science Foundation of Tianjin City(19JCYBJC28200)the Scientific Research Project of Tianjin Education Commission(2017KJ140)
文摘OBJECTIVE Shenmai Injection(SMI)is widely used in the treatment of cardiovascular diseases,such as heart failure and myocardial ischemia.In clinic,SMI showed protective effects on doxorubicin(Dox)-induced cardiac toxicity.In current study,we investigate the mitochondrial protective mechanisms of SMI on Dox-induced myocardial injury.METHODS C57BL/6 mice were divided into four groups:①control group;②Dox injury group;③SMI+Dox group and dexrazoxane(DRZ)+Dox group.Dex was a positive control.Myocardial injury was evaluated by echocardiography,HE and TUNEL staining,myocardial markers measurement.H9C2 cardiomyocytes pretreatment with SMI for 24 h were exposed to Dox.Cell viability and apoptosis were measured by CCK8,Hoechst33342 staining,and Annexin V/PI staining.MitoSOX,mitochondrial membrane potential,and mitochondrial respiratory function were measured to evaluate mito⁃chondrial function.RESULTS SMI decreased mortality rate of Dox-injected mice,serum CK and CK-MB levels in vivo.SMI significantly prevented Dox-induced cardiac dysfunction and apoptosis and increased expression level of PI3K,p-Akt,and p-GSK-3β.Moreover,SMI significantly inhibited Dox-induced apoptosis,mitochondrial ROS production,and reduction of mitochondrial membrane potential in H9C2 cells.Mechanismly,the cardio-protective effect of SMI was suppressed by PI3K inhibitor LY294002.CONCLUSION SMI prevents Dox-induced cardiotoxicity and mitochondrial damage through activation of PI3K/Akt signaling pathway.
