OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosap...OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosaponins(SS)-b on hepatocellular carcinoma(HCC)and its regulation on VEGF/ERK/HIF-1 αsignal pathway.METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro.Pathological change of tumor tissue was observed by HE staining,the microvascular changes were detected by immunohistochemical method.The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane(CAM)model.The effects of SS-b on proliferation,migration and invasion were investigated by MTT assay,scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell(HUVEC)and HepG2 cells in vitro.Vascular endothelial growth factor(VEGF),matrix metalloproteinase-2/9(MMP-2/9),hypoxia-inducible factor-1α(HIF-1α)expression and the phosphorylation of extracellular regulated kinase(ERK)were analyzed using RT-PCR and Westernblot.RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo.The inhibitory rate of tumor was 49.1%,50.7%,66.1%in SS-b 5,10 and 20 mg·kg-1group respectively.HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice.Moreover,SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF.SS-b had an obvious inhibitory effect on cell proliferation,migration and invasion of HUVEC cells and HepG-2 cells.These effects were associated with downregulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1αsignaling in H22 mice and Hep-G2 cells.CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibiting tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.展开更多
Endothelial progenitor cells(EPCs)are a circulating,bone marrow-derived cell population that participate in both vasculogenesis and vascular homeostasis.Recent studies have shown that EPCs are reduced by^50% in diabet...Endothelial progenitor cells(EPCs)are a circulating,bone marrow-derived cell population that participate in both vasculogenesis and vascular homeostasis.Recent studies have shown that EPCs are reduced by^50% in diabetes that correlates inversely with its mortality rate.In addition,EPC angiogenic functions are severely impaired in diabetes.However,the molecular and cellular mechanisms underlying EPC dysfunction are poorly understood.Our current studies have focused on in vitro and in展开更多
OBJECTIVE To analyse structure and relationship of several andrographolide derivatives in multiple in vivo and in vitro angiogenesis assays,and to demonstrate a novel compound named AGL-2as a potential anti-angiogenes...OBJECTIVE To analyse structure and relationship of several andrographolide derivatives in multiple in vivo and in vitro angiogenesis assays,and to demonstrate a novel compound named AGL-2as a potential anti-angiogenesis agent.METHODS Human umbilical vein endothelial cells(HUVECs)in vitro and zebrafish(Danio rerio)in vivo models were used to screen and identify the anti-angiogenesis activities of six andrographolide derivatives;namely,AGL-1,AGL-2,AGS-72,AGS-72 a,AGS-79 and AGP-151.RESULTS AGL-2exhibited the strongest anti-angiogenic activity among all the derivatives in zebrafish model.Interestingly,another compound named AGS-72 showed stronger anti-angiogenic activity than AGL-2 in VEGF-induced HUVECs proliferation,migration,invasion and tube formation assays.In addition,AGL-2 was found to suppress the VEGF-induced VEGFR-2auto-phosphorylation and inhibit the activity of VEGFR-2 mediated signaling cascades in a dose-dependent manner.CONCLUSION AGL-2was demonstrated to be a promising anti-angiogenic agent among all the tested derivatives.The mechanism underlying the anti-angiogenic activity of AGL-2 probably involve VEGFR-2 signaling pathway.Even though,how some of chemical structure alterations result in discrepancy between in vivo and in vitro activities still remains to be resolved,this study shall provide new insight into how modification of the chemical structure of andrographolide affects this newly identified anti-angiogenesis activity.Meanwhile,AGL-2 can be exploited as a potential therapeutic agent for the treatment of angiogenesis-related diseases.展开更多
OBJECTIVE Only limited number of drugs are currently available for treating ischemic stroke.Therapeutic angiogenesis has recently emerged as one of the most promising therapies for cerebral ischemic injury.Isopropyl-...OBJECTIVE Only limited number of drugs are currently available for treating ischemic stroke.Therapeutic angiogenesis has recently emerged as one of the most promising therapies for cerebral ischemic injury.Isopropyl-β-(3,4-dihydroxyphenyl)-α-hydroxypropanoate(IDHP)is a metabolite derived from the botanical formulation for Dantonic®.Here,we investigated the angiogenic efficacy of IDHP in cerebral ischemia.METHODS The in vivo effects of IDHP were evaluated in the C57BL/6 mouse Matrigel plug and rat transient middle cerebral artery occlusion(tMCAO)models.Primary human umbilical vein endothelial cells(HUVEC)and human brain microvascular endothelial cells(HBMEC)were used to explore the effects of IDHP on stimulating proliferation,migration and tube formation in vitro.ELISA and Western blotting were used to quantitate the release and expression of relevant target molecules and signaling pathways.RESULTS IDHP reduced infarct volume and improved sensorimotor function in rats subjected to tMCAO by promoting angiogenesis,and promoted Matrigel neovascularization in mice.Moreover,IDHP produced a biphasic modulation on proliferation and migration both in HUVEC and HBMEC.It also induced tube formation in a 12-day HUVEC-HDF co-culture model and in Matrigel assays.IDHP-induced angiogenesis was accompanied by increased levels of p-AMPKα(Thr172)and p-eNOS(Ser1177)both in vitro and in vivo,and the decreased level of VEGF in rat brains on day 1 whereas enhanced level of VEGF on day 3 and 7 after tMCAO.Mechanistically,AMPK knockdown or pharmacologically inhibiting AMPK and its upstream kinases(CaMKKβ)inhibited the eNOS phosphorylation induced by IDHP in HUVEC.Furthermore,selective eNOS inhibitor(L-NIO),selective CaMKKβinhibitor(STO)and AMPKa inhibitor(Compound C)blocked the capillary-like tube formation in the co-culture model induced by IDHP(10 nmol·L^(-1)).CONCLUSION Collectively,these findings showed that IDHP protected rats from cerebral ischemia-reperfusion injury by promoting angiogenesis via activating CaMKKβ/AMPK(Thr172)/eNOS(Ser1177)signaling,and suggest it to be a promising new drug candidate for the prevention and/or treatment of cerebral ischemia and other vascular occlusive diseases.展开更多
To investigate tumor angiogenesis under the influence of Endostatin,mathematical modeling and numerical simulation of tumor angiogenesis are performed,with the mechanical environment in matrix,the inhibiting effects o...To investigate tumor angiogenesis under the influence of Endostatin,mathematical modeling and numerical simulation of tumor angiogenesis are performed,with the mechanical environment in matrix,the inhibiting effects of Angiostatin and Endostatin into consideration.The展开更多
Despite significant progress in cancer research during the past decades,yet there are no major breakthroughs that can be translated into major benefits for the general public in terms of treatment or therapy for the c...Despite significant progress in cancer research during the past decades,yet there are no major breakthroughs that can be translated into major benefits for the general public in terms of treatment or therapy for the complex neoplastic diseases,especially for the malignant solid tumors.This depressing but indisputable fact leads to a call for new ideas to target tumor metastasis by editors of Nature Medicine<sup>[1]</sup>.The real problems are that the fundamental issues of transformation and malignancy in vivo are poorly understood.In a recent review on cancer,展开更多
Aim Baroreflex dysfunction is associated with a higher rate of sudden death after myocardial infarction （MI）. Ketanserin enhances baroreflex function in rats. The present work was designed to examine whether ketan- ...Aim Baroreflex dysfunction is associated with a higher rate of sudden death after myocardial infarction （MI）. Ketanserin enhances baroreflex function in rats. The present work was designed to examine whether ketan- serin improves the post-MI cardiac function and to explore the possible mechanism involved. Methods Spontane- ously hypertensive rats （SHR） were treated with ketanserin （0.3 mg · kg^-1 · d^-l）. Two weeks later, blood pres- sure and baroreflex function were measured, followed by a ligation of the left coronary artery. The expressions of ve- sicular acetylcholine transporter （VAChT） and α7 nicotinic acetylcholine receptor （α7-nAChR） in ischemic myo- cardium, angiogenesis, cardiac function, and left ventricular （LV） remodeling were evaluated subsequently. Re- sults Ketanserin significantly improved baroreflex sensitivity （0.62 ± 0. 21 vs. 0.34 ± 0. 12 ms/mmHg, P 〈 0.01 ） and vagal tonic activity （ heart rate changes in response to atropine, 54.8 ± 16.2 vs. 37.6 ± 13.4 b. p. m. , P 〈 0.01 ） without affecting the blood pressure or basic heart rate in SHR. Treatment of SHR with ketanserin promi- nently improved cardiac function and alleviated LV remodeling, as reflected by increases in the ejection fraction, fractional shortening, and LV systolic pressure as well as decreases in LV internal diameter and LV relative weight. The capillary density, vascular endothelial growth factor expression, and blood flow in the ischemic myocardium were significantly higher in the ketanserin-treated group. In addition, ketanserin markedly increased the expression of VAChT and α7-nAChR in ischemic myocardium. Conclusion Ketanserin improved post-MI cardiac function and angiogenesis in ischemic myocardium. The findings provide a mechanistic basis for restoring baroreflex function using ketanserin in the treatment of MI.展开更多
OBJECTIVE Uridine adenosine tetraphosphate(Up4A),a dinucleotide,contains both purine and pyrimidine moieties,and exerts its vascular influence via activation of purinergic receptors.Here,we aimed to investigate the ef...OBJECTIVE Uridine adenosine tetraphosphate(Up4A),a dinucleotide,contains both purine and pyrimidine moieties,and exerts its vascular influence via activation of purinergic receptors.Here,we aimed to investigate the effects of Up4 A on angiogenesis and the putative purinergic receptors(PR)involved in this process.METHODS Tubule formation assay was performed in 3D matrix system.In this assay,human umbilical vein endothelial cells(HUVECs)were co-cultured with pericytes with various Up4 A doses(0,1,2.5,5,10 and 20μmol·L-1)in the absence and presence of P2Y6 R antagonist MRS2578(10μmol·L-1)for 5d.Expression profile of PR subtypes and angiogenic factors was assessed in HUVECs by q-PCR with and without P2Y6 R antagonist.RESULTS No difference in initial tubule formation was detected between Up4 A stimulation and control conditions at day 2.In contrast,a significant increase in vascular density in response to Up4 A was observed at day 5.Up4 A at a dose of 2.5and 5μmol·L-1 promoted total tubule length(by-1.89 fold and-2.23fold),number of tubules(by-1.71 fold and-1.89fold)as well as number of junctions(by-2.24 fold and-2.80fold),all of which were inhibited by MRS2578.Further increase in Up4 A dose to10 and 20μmol·L-1 did not induce an increase in these vascular parameters as compared to non-treated controls.Moreover,Up4 A increased mRNA level of P2YRs(P2Y2R,P2Y4 R and P2Y6R)but not P2XR(P2X4R and P2X7R)or P1R(A2AR and A2BR),while Up4 A upregulated VEGFA and ANGPT1 but not VEGFR2,ANGPT2,Tie1 and Tie2at mRNA level.Transcriptional upregulation of P2 YRs and angiogenic factors by Up4 A was inhibited by MRS2578.CONCLUSION Up4 A is functionally capable of promoting tubule formation in vitro co-culture system.This process is likely mediated by activation of pyrimidine-favored P2 YRs but not P2 XR or P1 Rs,and involves stimulation of well known angiogenic factors.展开更多
基金supported by Scientific and Technology Projects of Henan Province(142102310137)Science and Technology Development Project of Luoyang City(1603001A-3)
文摘OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosaponins(SS)-b on hepatocellular carcinoma(HCC)and its regulation on VEGF/ERK/HIF-1 αsignal pathway.METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro.Pathological change of tumor tissue was observed by HE staining,the microvascular changes were detected by immunohistochemical method.The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane(CAM)model.The effects of SS-b on proliferation,migration and invasion were investigated by MTT assay,scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell(HUVEC)and HepG2 cells in vitro.Vascular endothelial growth factor(VEGF),matrix metalloproteinase-2/9(MMP-2/9),hypoxia-inducible factor-1α(HIF-1α)expression and the phosphorylation of extracellular regulated kinase(ERK)were analyzed using RT-PCR and Westernblot.RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo.The inhibitory rate of tumor was 49.1%,50.7%,66.1%in SS-b 5,10 and 20 mg·kg-1group respectively.HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice.Moreover,SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF.SS-b had an obvious inhibitory effect on cell proliferation,migration and invasion of HUVEC cells and HepG-2 cells.These effects were associated with downregulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1αsignaling in H22 mice and Hep-G2 cells.CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibiting tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.
基金Supported by Grants from NIH R01GM077352,VA Merit Award I01RX000244,AHA GIA0855601G,ADA Research Award7-08-RA-23NSFC Overseas Collaborative Grant#30728021
文摘Endothelial progenitor cells(EPCs)are a circulating,bone marrow-derived cell population that participate in both vasculogenesis and vascular homeostasis.Recent studies have shown that EPCs are reduced by^50% in diabetes that correlates inversely with its mortality rate.In addition,EPC angiogenic functions are severely impaired in diabetes.However,the molecular and cellular mechanisms underlying EPC dysfunction are poorly understood.Our current studies have focused on in vitro and in
基金The project supported by Science and Technology Development Fund of Macao SAR(078/2011/A3)Research Committee of University of Macao〔MYRG138(Y1-Y4)-ICMS12-LMY〕
文摘OBJECTIVE To analyse structure and relationship of several andrographolide derivatives in multiple in vivo and in vitro angiogenesis assays,and to demonstrate a novel compound named AGL-2as a potential anti-angiogenesis agent.METHODS Human umbilical vein endothelial cells(HUVECs)in vitro and zebrafish(Danio rerio)in vivo models were used to screen and identify the anti-angiogenesis activities of six andrographolide derivatives;namely,AGL-1,AGL-2,AGS-72,AGS-72 a,AGS-79 and AGP-151.RESULTS AGL-2exhibited the strongest anti-angiogenic activity among all the derivatives in zebrafish model.Interestingly,another compound named AGS-72 showed stronger anti-angiogenic activity than AGL-2 in VEGF-induced HUVECs proliferation,migration,invasion and tube formation assays.In addition,AGL-2 was found to suppress the VEGF-induced VEGFR-2auto-phosphorylation and inhibit the activity of VEGFR-2 mediated signaling cascades in a dose-dependent manner.CONCLUSION AGL-2was demonstrated to be a promising anti-angiogenic agent among all the tested derivatives.The mechanism underlying the anti-angiogenic activity of AGL-2 probably involve VEGFR-2 signaling pathway.Even though,how some of chemical structure alterations result in discrepancy between in vivo and in vitro activities still remains to be resolved,this study shall provide new insight into how modification of the chemical structure of andrographolide affects this newly identified anti-angiogenesis activity.Meanwhile,AGL-2 can be exploited as a potential therapeutic agent for the treatment of angiogenesis-related diseases.
基金National Natural Science Foundation of China(31971143)and Primary R&D Plan of Shaanxi Province(2021KWZ-24)。
文摘OBJECTIVE Only limited number of drugs are currently available for treating ischemic stroke.Therapeutic angiogenesis has recently emerged as one of the most promising therapies for cerebral ischemic injury.Isopropyl-β-(3,4-dihydroxyphenyl)-α-hydroxypropanoate(IDHP)is a metabolite derived from the botanical formulation for Dantonic®.Here,we investigated the angiogenic efficacy of IDHP in cerebral ischemia.METHODS The in vivo effects of IDHP were evaluated in the C57BL/6 mouse Matrigel plug and rat transient middle cerebral artery occlusion(tMCAO)models.Primary human umbilical vein endothelial cells(HUVEC)and human brain microvascular endothelial cells(HBMEC)were used to explore the effects of IDHP on stimulating proliferation,migration and tube formation in vitro.ELISA and Western blotting were used to quantitate the release and expression of relevant target molecules and signaling pathways.RESULTS IDHP reduced infarct volume and improved sensorimotor function in rats subjected to tMCAO by promoting angiogenesis,and promoted Matrigel neovascularization in mice.Moreover,IDHP produced a biphasic modulation on proliferation and migration both in HUVEC and HBMEC.It also induced tube formation in a 12-day HUVEC-HDF co-culture model and in Matrigel assays.IDHP-induced angiogenesis was accompanied by increased levels of p-AMPKα(Thr172)and p-eNOS(Ser1177)both in vitro and in vivo,and the decreased level of VEGF in rat brains on day 1 whereas enhanced level of VEGF on day 3 and 7 after tMCAO.Mechanistically,AMPK knockdown or pharmacologically inhibiting AMPK and its upstream kinases(CaMKKβ)inhibited the eNOS phosphorylation induced by IDHP in HUVEC.Furthermore,selective eNOS inhibitor(L-NIO),selective CaMKKβinhibitor(STO)and AMPKa inhibitor(Compound C)blocked the capillary-like tube formation in the co-culture model induced by IDHP(10 nmol·L^(-1)).CONCLUSION Collectively,these findings showed that IDHP protected rats from cerebral ischemia-reperfusion injury by promoting angiogenesis via activating CaMKKβ/AMPK(Thr172)/eNOS(Ser1177)signaling,and suggest it to be a promising new drug candidate for the prevention and/or treatment of cerebral ischemia and other vascular occlusive diseases.
基金supported by the National Natural Science Foundation of China,No.10372026,10772751Shanghai Leading Academic Discipline Project B112
文摘To investigate tumor angiogenesis under the influence of Endostatin,mathematical modeling and numerical simulation of tumor angiogenesis are performed,with the mechanical environment in matrix,the inhibiting effects of Angiostatin and Endostatin into consideration.The
基金supported by the funds from Huazhong University of Science and TechnologyUS NIH grant GM072744
文摘Despite significant progress in cancer research during the past decades,yet there are no major breakthroughs that can be translated into major benefits for the general public in terms of treatment or therapy for the complex neoplastic diseases,especially for the malignant solid tumors.This depressing but indisputable fact leads to a call for new ideas to target tumor metastasis by editors of Nature Medicine<sup>[1]</sup>.The real problems are that the fundamental issues of transformation and malignancy in vivo are poorly understood.In a recent review on cancer,
文摘Aim Baroreflex dysfunction is associated with a higher rate of sudden death after myocardial infarction （MI）. Ketanserin enhances baroreflex function in rats. The present work was designed to examine whether ketan- serin improves the post-MI cardiac function and to explore the possible mechanism involved. Methods Spontane- ously hypertensive rats （SHR） were treated with ketanserin （0.3 mg · kg^-1 · d^-l）. Two weeks later, blood pres- sure and baroreflex function were measured, followed by a ligation of the left coronary artery. The expressions of ve- sicular acetylcholine transporter （VAChT） and α7 nicotinic acetylcholine receptor （α7-nAChR） in ischemic myo- cardium, angiogenesis, cardiac function, and left ventricular （LV） remodeling were evaluated subsequently. Re- sults Ketanserin significantly improved baroreflex sensitivity （0.62 ± 0. 21 vs. 0.34 ± 0. 12 ms/mmHg, P 〈 0.01 ） and vagal tonic activity （ heart rate changes in response to atropine, 54.8 ± 16.2 vs. 37.6 ± 13.4 b. p. m. , P 〈 0.01 ） without affecting the blood pressure or basic heart rate in SHR. Treatment of SHR with ketanserin promi- nently improved cardiac function and alleviated LV remodeling, as reflected by increases in the ejection fraction, fractional shortening, and LV systolic pressure as well as decreases in LV internal diameter and LV relative weight. The capillary density, vascular endothelial growth factor expression, and blood flow in the ischemic myocardium were significantly higher in the ketanserin-treated group. In addition, ketanserin markedly increased the expression of VAChT and α7-nAChR in ischemic myocardium. Conclusion Ketanserin improved post-MI cardiac function and angiogenesis in ischemic myocardium. The findings provide a mechanistic basis for restoring baroreflex function using ketanserin in the treatment of MI.
文摘OBJECTIVE Uridine adenosine tetraphosphate(Up4A),a dinucleotide,contains both purine and pyrimidine moieties,and exerts its vascular influence via activation of purinergic receptors.Here,we aimed to investigate the effects of Up4 A on angiogenesis and the putative purinergic receptors(PR)involved in this process.METHODS Tubule formation assay was performed in 3D matrix system.In this assay,human umbilical vein endothelial cells(HUVECs)were co-cultured with pericytes with various Up4 A doses(0,1,2.5,5,10 and 20μmol·L-1)in the absence and presence of P2Y6 R antagonist MRS2578(10μmol·L-1)for 5d.Expression profile of PR subtypes and angiogenic factors was assessed in HUVECs by q-PCR with and without P2Y6 R antagonist.RESULTS No difference in initial tubule formation was detected between Up4 A stimulation and control conditions at day 2.In contrast,a significant increase in vascular density in response to Up4 A was observed at day 5.Up4 A at a dose of 2.5and 5μmol·L-1 promoted total tubule length(by-1.89 fold and-2.23fold),number of tubules(by-1.71 fold and-1.89fold)as well as number of junctions(by-2.24 fold and-2.80fold),all of which were inhibited by MRS2578.Further increase in Up4 A dose to10 and 20μmol·L-1 did not induce an increase in these vascular parameters as compared to non-treated controls.Moreover,Up4 A increased mRNA level of P2YRs(P2Y2R,P2Y4 R and P2Y6R)but not P2XR(P2X4R and P2X7R)or P1R(A2AR and A2BR),while Up4 A upregulated VEGFA and ANGPT1 but not VEGFR2,ANGPT2,Tie1 and Tie2at mRNA level.Transcriptional upregulation of P2 YRs and angiogenic factors by Up4 A was inhibited by MRS2578.CONCLUSION Up4 A is functionally capable of promoting tubule formation in vitro co-culture system.This process is likely mediated by activation of pyrimidine-favored P2 YRs but not P2 XR or P1 Rs,and involves stimulation of well known angiogenic factors.
