Angiogenesis in the infarct periphery can improve blood flow. Vascular endothelial growth factor （VEGF） has been considered a potential therapeutic target for stroke. Buyang Huanwu decoction （BYHWD） is a classic t...Angiogenesis in the infarct periphery can improve blood flow. Vascular endothelial growth factor （VEGF） has been considered a potential therapeutic target for stroke. Buyang Huanwu decoction （BYHWD） is a classic traditional formula in traditional Chinese medicine and is used to treat stroke; in addition, the promotion effects on VEGF protein expression have been confirmed. However, little is known about how BYHWD regulates angiogenesis, or about the effects of BYHWD on VEGF mRNA expression. For this reason, the present study measured microvessel density in rats with cerebral ischemia using immunohistochemistry. In addition, VEGF expression was measured by re-verse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay to determine the effects of BYHWD on angiogenesis and VEGF expression in rats with cerebral ischemia. Results demonstrated that microvessel density, as well as VEGF mRNA and protein expression, increased after 7 and 14 days of BYHWD treatment, which suggests that BYHWD promoted angiogenesis following cerebral ischemia and upregulated VEGF mRNA and protein expression in ischemic cerebral regions.展开更多
BACKGROUND： Therapeutic angiogenesis has opened up new pathway for the treatment of ischemic cerebrovascular disease in recent years. The exploration of the effect of vascular endothelial growth factor （VEGF） on in...BACKGROUND： Therapeutic angiogenesis has opened up new pathway for the treatment of ischemic cerebrovascular disease in recent years. The exploration of the effect of vascular endothelial growth factor （VEGF） on inducing angiogenesis following ischemia/reperfusion injury can provide better help for the long-term treatment of cerebrovascular disease in clinic. OBJECTIVE： To observe the effect of VEGF on inducing angiogenesis following focal cerebral ischemia /reperfusion injury in rabbits through the angiogenesis of microvessels reflected by the expression of the factors of vascular pseudohemophilia. DESIGN： A randomized controlled animal tria SETTNG： Department of Medical Imaging, Second Hospital of Hebei Medical University MATERIALS： Sixty-five healthy male New Zealand rabbits of clean degree, weighing （2.6±0.2） kg, aged 4.5-5 months, were used. The polyclonal antibody against vascular pseudohemophilia （Beijing Zhongshan Company）, recombinant VEGF165 （Peprotech Company, USA）, biotinylated second antibody and ABC compound （Wuhan Boster Company） were applied. METHODS： The experiments were carried out in the Laboratory of Neuromolecular Imaging and Neuropathy, Second Hospital of Hebei Medical University from May to August in 2005. （1） The rabbits were randomly divided into three groups： sham-operated group （n=15）, control group （n=25） and VEGF-treated group （n=-25）. In the control group and VEGF-treated group, models were established by middle cerebral artery occlusion （MCAO） induced focal cerebral ischemia/reperfusion. In the VEGF-treated group, VEGF165 （2.5 mg/L） was stereotactically injected into the surrounding regions of the infarcted sites immediately after the 2-hour ischemia/reperfusion; Saline of the same dosage was injected in the control group. But the rabbits in the sham-operated group were only drilled but not administrated. （2） The experimental indexes were observed on the 3^rd 7^th, 14^th, 28^th and 70^th days of the experiment respectively, 3 rabbits in the sham-operated group and 5 in the control group and VEGF-treated group were observed at each time point. The brain tissues in the surrounding regions of the infarcted sites were collected. The positive expressions of the factors of vascular pseudohemophilia in vascular endothelial cells were analyzed with immunohistochemical method. The microvessels in unit statistical field were counted with the imaging analytical software. MAIN OUTCOME MEASURES： The changes of microvascular density in the brain tissue and the positive expressions of the factors of vascular pseudohemophilia in the surrounding regions of the infarcted sites were observed on the 3^rd 7^th, 14^th, 28^th and 70^th days of the experiment. RESULTS： All the 65 New Zealand rabbits were involved in the analysis of results without deletion. Changes of the number of microvessels at different time points in each group： There were no obvious changes at different time points in the sham-operated group. The numbers of microvessels at 7 and 14 days were obviously more in the control group than in the sham-operated group [（6.0±1.1）, （9.0±0.9） microvessels; （3.0±1.1）, （3.0±1.1） microvessels; P〈 0.05-0.01], and those at 3, 7, 14 and 28 days were obviously more in the VEGF-treated group than in the control group [（8.3±2.0）, （13.4±1.4）, （15.5±2.3）, （6.8± 1.0） microvessels; （3.4±0.6）, （6.0±1.1）, （9.0±0.9）, （3.2±0.8） microvessels; P 〈 0.01]. （2） Positive expressions of the factors of vascular pseudohemophilia in the surrounding regions of infarcted sites： There were no obvious changes at different time points in the sham-operated group. In the control group, the changing law of the expressions was the same as that for the number of microvessels that the expression began to mildly increase at 7 days, reached the peak value at 14 days, and began to reduce at 28 days. In the VEGF-treated group, the expression was obviously increased at 3 days, also reached the peak value at 14 days, and reduced to the normal level at 70 days, but the expressions were obviously stronger than those in the control group at the same time points. CONCLUSION： Angiogenesis can be obviously induced in rabbits after the focal cerebral ischemia/reperfusion injury is treated with VEGF for 18 days.展开更多
Buyanghuanwu decoction has been shown to protect against cerebral ischemia/reperfusion injury,but the underlying mechanisms remain unclear.In this study,rats were intragastrically given Buyanghuanwu decoction,15 m L/k...Buyanghuanwu decoction has been shown to protect against cerebral ischemia/reperfusion injury,but the underlying mechanisms remain unclear.In this study,rats were intragastrically given Buyanghuanwu decoction,15 m L/kg,for 3 days.A rat model of cerebral ischemia/reperfusion injury was established by middle cerebral artery occlusion.In rats administered Buyanghuanwu decoction,infarct volume was reduced,serum vascular endothelial growth factor and integrin αvβ3 levels were increased,and brain tissue vascular endothelial growth factor and CD34 expression levels were increased compared with untreated animals.These effects of Buyanghuanwu decoction were partially suppressed by an angiogenesis inhibitor（administered through the lateral ventricle for 7 consecutive days）.These data suggest that Buyanghuanwu decoction promotes angiogenesis,improves cerebral circulation,and enhances brain tissue repair after cerebral ischemia/reperfusion injury.展开更多
Cerebral angiogenesis in the early stages after cerebral ischemia injury is essential for the recovery of nerve function,in which fetal liver kinase-1（Flk-1）,as a major regulator of vasculogenesis and angiogenesis,p...Cerebral angiogenesis in the early stages after cerebral ischemia injury is essential for the recovery of nerve function,in which fetal liver kinase-1（Flk-1）,as a major regulator of vasculogenesis and angiogenesis,plays a very important role.Microvessel density（MVD）was greater in an aged model group compared with the young sham operated group（P 〈 0.01）.MVD and the sum of the lumen area were decreased in the aged group at 1,3,6 and 12 days following ischemia/reperfusion（I/R）injury compared with the young model group（P 〈 0.05 and P 〈 0.01,respectively）.Flk-1 protein and mRNA expression was greater in the aged model group when compared with the young sham operated group（P 〈 0.01）.Flk-1 protein and mRNA expression was lower in the aged group at 1,3,6 and 12 days after I/R compared with the young model group（P 〈 0.01）.Flk-1 expression in aged rats attenuated rapidly,but was still maintained at relatively higher levels at 12 days following I/R in younger rats.The results suggest that angiogenesis was weakened after cerebral I/R in aged rats,and the mechanism of which might be correlated with attenuated expression of Flk-1 protein and mRNA.展开更多
OBJECTIVE:To investigate the combinatorial effects of Naomai Yihao(NMYH) Capsules(脑脉一号胶囊) and vascular endothelial growth factor(VEGF) gene-transfected bone marrow mesenchymal stem cells(BMSCs) on angiogenesis i...OBJECTIVE:To investigate the combinatorial effects of Naomai Yihao(NMYH) Capsules(脑脉一号胶囊) and vascular endothelial growth factor(VEGF) gene-transfected bone marrow mesenchymal stem cells(BMSCs) on angiogenesis in cerebral ischemic tissues in rats and the mechanism.METHOD:BMSCs were isolated and cultured from bone marrow by an adherence method.Then,BMSCs were transfected with the eukaryotic expression plasmid pEGFP-VEGF 165 by positive ionic liposome transfection.A rat model of middle cerebral artery occlusion(MCAO) was established.Rats were allocated to six groups:model,BMSC,VEGF gene-transfected BMSC transplantation(BMSC/VEGF),NMYH,combined NMYH and BMSC/VEGF(combined treatment group) and sham operation groups.The behavioral rating score(BRS) of rat and the expression of CD34 and VEGF in brain tis sue were measured by immunohistochemistry on days 7,14 and 21 after reperfusion.Angiogenesi was observed and evaluated with laser scanning confocal microscopy.RESULTS:The BRS of rats in NMYH,BMSC transplan tation and combined treatment groups was significantly lower than that of the model group(P< 0.001),with no significant difference between NMYH and transplantation groups(P=0.619).The expression of CD34 andVEGF in NMYH,transplanta tion and combined treatment groups increased(P< 0.001),with a significant difference between NMYH and transplantation groups(P<0.001).The blood vessel area in NMYH,transplantation and com bined treatment groups was significantly increased(P<0.05),without a significant difference between NMYH and transplantation groups(P=0.873).CONCLUSIONS:VEGF gene-transfected BMSCs im prove angiogenesis in the cerebral ischemic area NMYH Capsules promote angiogenesis in MCAO rats treated with BMSC transplantation,which show an improved BRS.The mechanism of angio genesis may be related to up-regulation ofVEGF ex pression.展开更多
Mesenchymal stem cell transplantation is a novel means of treating cerebral ischemia/reper- fusion, and can promote angiogenesis and neurological functional recovery. Acupuncture at Conception and Governor vessels als...Mesenchymal stem cell transplantation is a novel means of treating cerebral ischemia/reper- fusion, and can promote angiogenesis and neurological functional recovery. Acupuncture at Conception and Governor vessels also has positive effects as a treatment for cerebral ischemia/ reperfusion. Therefore, we hypothesized that electro-acupuncture at Conception and Governor vessels plus mesenchymal stem cell transplantation may have better therapeutic effects on the promotion of angiogenesis and recovery of neurological function than either treatment alone. In the present study, human umbilical cord blood-derived mesenchymal stem cells were isolated, cultured, identified and intracranially transplanted into the striatum and subcortex of rats at 24 hours following cerebral ischemia/reperfusion. Subsequently, rats were electro-acupunctured at Conception and Governor vessels at 24 hours after transplantation. Modified neurological severity scores and immunohistochemistry findings revealed that the combined interventions of electro-acupuncture and mesenchymal stem cell transplantation clearly improved neurological impairment and up-regulated vascular endothelial growth factor expression around the isch- emic focus. The combined intervention provided a better outcome than mesenchymal stem cell transplantation alone. These findings demonstrate that electro-acupuncture at Conception and Governor vessels and mesenchymal stem cell transplantation have synergetic effects on promot- ing neurological function recovery and angiogenesis in rats after cerebral ischemia/reperfusion.展开更多
Objective To explore the changes of endostatin （a strong anti-angiogenesis factor） and vascular endothelial growth factor （VEGF） in the brain tissues of rabbits following cerebral ischemia induced by middle cerebr...Objective To explore the changes of endostatin （a strong anti-angiogenesis factor） and vascular endothelial growth factor （VEGF） in the brain tissues of rabbits following cerebral ischemia induced by middle cerebral artery occlusion （MCAO）. Methods Twenty-four New Zealand white rabbits were randomly divided into 5 groups： control （n = 5）, sham-operation （n = 4）, 2-hour ischemia （n = 5）, 24-hour ischemia （n = 5）, and 48-hour ischemia （n = 5）. The expression of VEGF and endostatin were measured by enzyme-linked immunosorbent assay （ELISA） and immunohistochemistry, respectively. In situ hybridization was used to characterize the expression of mRNA for the endostatin. Results Both the protein （at least 50%, P 〈 0.01） and mRNA （at least 70%, P 〈 0.05） of endostatin increased significantly in the ischemic brain tissues after MCAO compared with the control group. VEGF increased at least 270% in the brain after cerebral ischemia （P 〈 0.05）. Conclusion Cerebral ischemia leads to an up-regulation of endostatin in the brain, which is not associated with the increase of VEGF in the brain. The increase of endostatin may serve as a deleterious mechanism for ischemic injury through blocking angiogenesis.展开更多
基金the National Natural Science Foundation of China,No.30300470,30472217,30873355the Key Project of Chinese Ministry Education of China,No.209087+1 种基金the Key Project of Natural Science Foundation of Hunan Province,No.06JJ2052a Grant from the Educational Commission of Hunan Province,No.08A049
文摘Angiogenesis in the infarct periphery can improve blood flow. Vascular endothelial growth factor （VEGF） has been considered a potential therapeutic target for stroke. Buyang Huanwu decoction （BYHWD） is a classic traditional formula in traditional Chinese medicine and is used to treat stroke; in addition, the promotion effects on VEGF protein expression have been confirmed. However, little is known about how BYHWD regulates angiogenesis, or about the effects of BYHWD on VEGF mRNA expression. For this reason, the present study measured microvessel density in rats with cerebral ischemia using immunohistochemistry. In addition, VEGF expression was measured by re-verse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay to determine the effects of BYHWD on angiogenesis and VEGF expression in rats with cerebral ischemia. Results demonstrated that microvessel density, as well as VEGF mRNA and protein expression, increased after 7 and 14 days of BYHWD treatment, which suggests that BYHWD promoted angiogenesis following cerebral ischemia and upregulated VEGF mRNA and protein expression in ischemic cerebral regions.
文摘BACKGROUND： Therapeutic angiogenesis has opened up new pathway for the treatment of ischemic cerebrovascular disease in recent years. The exploration of the effect of vascular endothelial growth factor （VEGF） on inducing angiogenesis following ischemia/reperfusion injury can provide better help for the long-term treatment of cerebrovascular disease in clinic. OBJECTIVE： To observe the effect of VEGF on inducing angiogenesis following focal cerebral ischemia /reperfusion injury in rabbits through the angiogenesis of microvessels reflected by the expression of the factors of vascular pseudohemophilia. DESIGN： A randomized controlled animal tria SETTNG： Department of Medical Imaging, Second Hospital of Hebei Medical University MATERIALS： Sixty-five healthy male New Zealand rabbits of clean degree, weighing （2.6±0.2） kg, aged 4.5-5 months, were used. The polyclonal antibody against vascular pseudohemophilia （Beijing Zhongshan Company）, recombinant VEGF165 （Peprotech Company, USA）, biotinylated second antibody and ABC compound （Wuhan Boster Company） were applied. METHODS： The experiments were carried out in the Laboratory of Neuromolecular Imaging and Neuropathy, Second Hospital of Hebei Medical University from May to August in 2005. （1） The rabbits were randomly divided into three groups： sham-operated group （n=15）, control group （n=25） and VEGF-treated group （n=-25）. In the control group and VEGF-treated group, models were established by middle cerebral artery occlusion （MCAO） induced focal cerebral ischemia/reperfusion. In the VEGF-treated group, VEGF165 （2.5 mg/L） was stereotactically injected into the surrounding regions of the infarcted sites immediately after the 2-hour ischemia/reperfusion; Saline of the same dosage was injected in the control group. But the rabbits in the sham-operated group were only drilled but not administrated. （2） The experimental indexes were observed on the 3^rd 7^th, 14^th, 28^th and 70^th days of the experiment respectively, 3 rabbits in the sham-operated group and 5 in the control group and VEGF-treated group were observed at each time point. The brain tissues in the surrounding regions of the infarcted sites were collected. The positive expressions of the factors of vascular pseudohemophilia in vascular endothelial cells were analyzed with immunohistochemical method. The microvessels in unit statistical field were counted with the imaging analytical software. MAIN OUTCOME MEASURES： The changes of microvascular density in the brain tissue and the positive expressions of the factors of vascular pseudohemophilia in the surrounding regions of the infarcted sites were observed on the 3^rd 7^th, 14^th, 28^th and 70^th days of the experiment. RESULTS： All the 65 New Zealand rabbits were involved in the analysis of results without deletion. Changes of the number of microvessels at different time points in each group： There were no obvious changes at different time points in the sham-operated group. The numbers of microvessels at 7 and 14 days were obviously more in the control group than in the sham-operated group [（6.0±1.1）, （9.0±0.9） microvessels; （3.0±1.1）, （3.0±1.1） microvessels; P〈 0.05-0.01], and those at 3, 7, 14 and 28 days were obviously more in the VEGF-treated group than in the control group [（8.3±2.0）, （13.4±1.4）, （15.5±2.3）, （6.8± 1.0） microvessels; （3.4±0.6）, （6.0±1.1）, （9.0±0.9）, （3.2±0.8） microvessels; P 〈 0.01]. （2） Positive expressions of the factors of vascular pseudohemophilia in the surrounding regions of infarcted sites： There were no obvious changes at different time points in the sham-operated group. In the control group, the changing law of the expressions was the same as that for the number of microvessels that the expression began to mildly increase at 7 days, reached the peak value at 14 days, and began to reduce at 28 days. In the VEGF-treated group, the expression was obviously increased at 3 days, also reached the peak value at 14 days, and reduced to the normal level at 70 days, but the expressions were obviously stronger than those in the control group at the same time points. CONCLUSION： Angiogenesis can be obviously induced in rabbits after the focal cerebral ischemia/reperfusion injury is treated with VEGF for 18 days.
基金financially supported by the National Natural Science Foundation of China,No.81072799
文摘Buyanghuanwu decoction has been shown to protect against cerebral ischemia/reperfusion injury,but the underlying mechanisms remain unclear.In this study,rats were intragastrically given Buyanghuanwu decoction,15 m L/kg,for 3 days.A rat model of cerebral ischemia/reperfusion injury was established by middle cerebral artery occlusion.In rats administered Buyanghuanwu decoction,infarct volume was reduced,serum vascular endothelial growth factor and integrin αvβ3 levels were increased,and brain tissue vascular endothelial growth factor and CD34 expression levels were increased compared with untreated animals.These effects of Buyanghuanwu decoction were partially suppressed by an angiogenesis inhibitor（administered through the lateral ventricle for 7 consecutive days）.These data suggest that Buyanghuanwu decoction promotes angiogenesis,improves cerebral circulation,and enhances brain tissue repair after cerebral ischemia/reperfusion injury.
基金the National Natural Science Foundation of China, No. 30371812the Henan Provincial Science Fund for Distinguished Young Scholars, No. 0612000700
文摘Cerebral angiogenesis in the early stages after cerebral ischemia injury is essential for the recovery of nerve function,in which fetal liver kinase-1（Flk-1）,as a major regulator of vasculogenesis and angiogenesis,plays a very important role.Microvessel density（MVD）was greater in an aged model group compared with the young sham operated group（P 〈 0.01）.MVD and the sum of the lumen area were decreased in the aged group at 1,3,6 and 12 days following ischemia/reperfusion（I/R）injury compared with the young model group（P 〈 0.05 and P 〈 0.01,respectively）.Flk-1 protein and mRNA expression was greater in the aged model group when compared with the young sham operated group（P 〈 0.01）.Flk-1 protein and mRNA expression was lower in the aged group at 1,3,6 and 12 days after I/R compared with the young model group（P 〈 0.01）.Flk-1 expression in aged rats attenuated rapidly,but was still maintained at relatively higher levels at 12 days following I/R in younger rats.The results suggest that angiogenesis was weakened after cerebral I/R in aged rats,and the mechanism of which might be correlated with attenuated expression of Flk-1 protein and mRNA.
基金Supported by the Research Fund for the Doctoral Program of Higher Education of China (No.20070572004,20104425120009)Guangdong Natural Science Fund(No.06301402)Traditional Chinese Medicine Master Education Program of Tongji University (Sponsored by State Ad-ministration of Traditional Chinese Medicine of the People'sRepublic of China,No:[2008]185)
文摘OBJECTIVE:To investigate the combinatorial effects of Naomai Yihao(NMYH) Capsules(脑脉一号胶囊) and vascular endothelial growth factor(VEGF) gene-transfected bone marrow mesenchymal stem cells(BMSCs) on angiogenesis in cerebral ischemic tissues in rats and the mechanism.METHOD:BMSCs were isolated and cultured from bone marrow by an adherence method.Then,BMSCs were transfected with the eukaryotic expression plasmid pEGFP-VEGF 165 by positive ionic liposome transfection.A rat model of middle cerebral artery occlusion(MCAO) was established.Rats were allocated to six groups:model,BMSC,VEGF gene-transfected BMSC transplantation(BMSC/VEGF),NMYH,combined NMYH and BMSC/VEGF(combined treatment group) and sham operation groups.The behavioral rating score(BRS) of rat and the expression of CD34 and VEGF in brain tis sue were measured by immunohistochemistry on days 7,14 and 21 after reperfusion.Angiogenesi was observed and evaluated with laser scanning confocal microscopy.RESULTS:The BRS of rats in NMYH,BMSC transplan tation and combined treatment groups was significantly lower than that of the model group(P< 0.001),with no significant difference between NMYH and transplantation groups(P=0.619).The expression of CD34 andVEGF in NMYH,transplanta tion and combined treatment groups increased(P< 0.001),with a significant difference between NMYH and transplantation groups(P<0.001).The blood vessel area in NMYH,transplantation and com bined treatment groups was significantly increased(P<0.05),without a significant difference between NMYH and transplantation groups(P=0.873).CONCLUSIONS:VEGF gene-transfected BMSCs im prove angiogenesis in the cerebral ischemic area NMYH Capsules promote angiogenesis in MCAO rats treated with BMSC transplantation,which show an improved BRS.The mechanism of angio genesis may be related to up-regulation ofVEGF ex pression.
基金supported by the National Natural Science Foundation of China,No.81072877Key Laboratory Project of Condition and Platform Construction Plan of Shenzhen Scientific Research Fund,No.CXB201111250113AShenzhen Scientific and Technology Development Program,No.201203149
文摘Mesenchymal stem cell transplantation is a novel means of treating cerebral ischemia/reper- fusion, and can promote angiogenesis and neurological functional recovery. Acupuncture at Conception and Governor vessels also has positive effects as a treatment for cerebral ischemia/ reperfusion. Therefore, we hypothesized that electro-acupuncture at Conception and Governor vessels plus mesenchymal stem cell transplantation may have better therapeutic effects on the promotion of angiogenesis and recovery of neurological function than either treatment alone. In the present study, human umbilical cord blood-derived mesenchymal stem cells were isolated, cultured, identified and intracranially transplanted into the striatum and subcortex of rats at 24 hours following cerebral ischemia/reperfusion. Subsequently, rats were electro-acupunctured at Conception and Governor vessels at 24 hours after transplantation. Modified neurological severity scores and immunohistochemistry findings revealed that the combined interventions of electro-acupuncture and mesenchymal stem cell transplantation clearly improved neurological impairment and up-regulated vascular endothelial growth factor expression around the isch- emic focus. The combined intervention provided a better outcome than mesenchymal stem cell transplantation alone. These findings demonstrate that electro-acupuncture at Conception and Governor vessels and mesenchymal stem cell transplantation have synergetic effects on promot- ing neurological function recovery and angiogenesis in rats after cerebral ischemia/reperfusion.
文摘Objective To explore the changes of endostatin （a strong anti-angiogenesis factor） and vascular endothelial growth factor （VEGF） in the brain tissues of rabbits following cerebral ischemia induced by middle cerebral artery occlusion （MCAO）. Methods Twenty-four New Zealand white rabbits were randomly divided into 5 groups： control （n = 5）, sham-operation （n = 4）, 2-hour ischemia （n = 5）, 24-hour ischemia （n = 5）, and 48-hour ischemia （n = 5）. The expression of VEGF and endostatin were measured by enzyme-linked immunosorbent assay （ELISA） and immunohistochemistry, respectively. In situ hybridization was used to characterize the expression of mRNA for the endostatin. Results Both the protein （at least 50%, P 〈 0.01） and mRNA （at least 70%, P 〈 0.05） of endostatin increased significantly in the ischemic brain tissues after MCAO compared with the control group. VEGF increased at least 270% in the brain after cerebral ischemia （P 〈 0.05）. Conclusion Cerebral ischemia leads to an up-regulation of endostatin in the brain, which is not associated with the increase of VEGF in the brain. The increase of endostatin may serve as a deleterious mechanism for ischemic injury through blocking angiogenesis.
