Blood vessels constitute a closed pipe system distributed throughout the body,transporting blood from the heart to other organs and delivering metabolic waste products back to the lungs and kidneys.Changes in blood ve...Blood vessels constitute a closed pipe system distributed throughout the body,transporting blood from the heart to other organs and delivering metabolic waste products back to the lungs and kidneys.Changes in blood vessels are related to many disorders like stroke,myocardial infarction,aneurysm,and diabetes,which are important causes of death worldwide.Translational research for new appro-aches to disease modeling and effective treatment is needed due to the huge socio-economic burden on healthcare systems.Although mice or rats have been widely used,applying data from animal studies to human-specific vascular physiology and pathology is difficult.The rise of induced pluripotent stem cells(iPSCs)provides a reliable in vitro resource for disease modeling,regenerative medicine,and drug discovery because they carry all human genetic information and have the ability to directionally differentiate into any type of human cells.This review summarizes the latest progress from the establishment of iPSCs,the strategies for differentiating iPSCs into vascular cells,and the in vivo trans-plantation of these vascular derivatives.It also introduces the application of these technologies in disease modeling,drug screening,and regenerative medicine.Additionally,the application of high-tech tools,such as omics analysis and high-throughput sequencing,in this field is reviewed.展开更多
Endothelial cells (TEC3 cells) derived from mouse embryonic stem (ES) cells were used as seed cells to construct blood vessels. Tissue engineered blood vessels were made by seeding 8 × l06 smooth muscle cells (SM...Endothelial cells (TEC3 cells) derived from mouse embryonic stem (ES) cells were used as seed cells to construct blood vessels. Tissue engineered blood vessels were made by seeding 8 × l06 smooth muscle cells (SMCs) obtained from rabbit arteries onto a sheet of nonwoven polyglycolic acid (PGA) fibers, which was used as a biodegradable polymer scaffold. After being cultured in DMEM medium for 7 days in vitro, SMCs grew well on the PGA fibers, and the cell-PGA sheet was then wrapped around a silicon tube, and implanted subcutaneously into nude mice. After 6~8 weeks, the silicon tube was replaced with another silicon tube in smaller diameter, and then the TEC3 cells (endothelial cells differentiated from mouse ES cells) were injected inside the engineered vessel tube as the test group. In the control group only culture medium was injected. Five days later, the engineered vessels were harvested for gross observation, histological and immunohistochemical analysis. The preliminary results demonstrated that the SMC-PGA construct could form a tubular structure in 6~8 weeks and PGA fibers were completely degraded. Histological and immunohistochemical analysis of the newly formed tissue revealed a typical blood vessel structure, including a lining of endothelial cells (ECs) on the lumimal surface and the presence of SMC and collagen in the wall. No EC lining was found in the tubes of control group. Therefore, the ECs differentiated from mouse ES cells can serve as seed cells for endothelium lining in tissue engineered blood vessels.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
The coronavirus disease 2019(COVID-19)infected so far over 250 million people and caused the death of over 5 million worldwide.Aging,diabetes,and cardiovascular diseases,conditions with preexisting impaired endothelia...The coronavirus disease 2019(COVID-19)infected so far over 250 million people and caused the death of over 5 million worldwide.Aging,diabetes,and cardiovascular diseases,conditions with preexisting impaired endothelial functions predispose to COVID-19.While respiratory epithelium is the main route of virus entry,the endothelial cells(ECs)lining pulmonary blood vessels are also an integral part of lung injury in COVID-19 patients.COVID-19 not only affects the lungs and respiratory system but also gastrointestinal(GI)tract,liver,pancreas,kidneys,heart,brain,and skin.Blood vessels are likely conduits for the virus dissemination to these distant organs.Importantly,ECs are also critical for vascular regeneration during injury/lesions healing and restoration of vascular network.The World Journal of Gastroenterology has published in last two years over 67 outstanding papers on COVID-19 infection with a focus on the GI tract,liver,pancreas,etc.,however,the role of the endothelial and vascular components as major targets for COVID-19-induced tissue injury,spreading to various organs,and injury healing have not been sufficiently emphasized.In the present article,we focus on these subjects and on current treatments including the most recent oral drugs molnupiravir and paxlovid that show a dramatic,significant efficacy in controlling severe COVID-19 infection.展开更多
Selenoprotein biosynthesis may not only be affected by the availability of selenium and the transcription rate of pertinent genes but also by the activity of components of the selenocysteine incorporation complex, Sel...Selenoprotein biosynthesis may not only be affected by the availability of selenium and the transcription rate of pertinent genes but also by the activity of components of the selenocysteine incorporation complex, SelA, B, C, or D. Incorporation of selenocysteine into selenoproteins requires a complex co-translational mechanism guaranteeing the correct recoding of the termination codon TGA as selenocysteine codon. A particular tRNASer(Sec) is enzyrnatically transformed by selenophosphate into tRNAsec which recognizes the UGA codon by means of a specific elongation factor (SelB) and a peculiar mRNA secondary structure. Selenophosphate is formed from selenide and ATP by the SelD gene product, selenophosphate synthase (SelD). To further elucidate the biological role of phospholipid hydroperoxide GPx (PHGPx), we transformed cells with a heterologous (pig) PHGPx gene and/or an additional (human) SelD gene and studied the behaviour of these cells under selenium depletion and repletion. Transfection of the endothelial cell line ECV 304 with either PHGPx cDNA or SelD cDNA did not result in a substantial increase of PHGPx activities, independent of selenium supply. However, cells co-trans fected with both, PHGPx and SelD cDNA, expressed significantly higher PHGPx activlty. This effect was much more pronounced under selenium limiting conditions. The enhanced PHGPx activity correlated with two functional pararneters, increased capability to reduce hydroperoxides and less sensitivity against H2O2-induced cytotoxicity. Thus, the ECV cells, stably transfected with PHGPx and SelD cDNA, provide a model to specifically investigate the role of PHGPx in endothelial cell function展开更多
The blood-brain barrier is a unique property of central nervous system blood vessels that protects sensitive central nervous system cells from potentially harmful blood components.The mechanistic basis of this barrier...The blood-brain barrier is a unique property of central nervous system blood vessels that protects sensitive central nervous system cells from potentially harmful blood components.The mechanistic basis of this barrier is found at multiple levels,including the adherens and tight junction proteins that tightly bind adjacent endothelial cells and the influence of neighboring pericytes,microglia,and astrocyte endfeet.In addition,extracellular matrix components of the vascular basement membrane play a critical role in establishing and maintaining blood-brain barrier integrity,not only by providing an adhesive substrate for blood-brain barrier cells to adhere to,but also by providing guidance cues that strongly influence vascular cell behavior.The extracellular matrix protein laminin is one of the most abundant components of the basement membrane,and several lines of evidence suggest that it plays a key role in directing blood-brain barrier behavior.In this review,we describe the basic structure of laminin and its receptors,the expression patterns of these molecules in central nervous system blood vessels and how they are altered in disease states,and most importantly,how genetic deletion of different laminin isoforms or their receptors reveals the contribution of these molecules to blood-brain barrier function and integrity.Finally,we discuss some of the important unanswered questions in the field and provide a“to-do”list of some of the critical outstanding experiments.展开更多
Purpose:To investigate the spatial and temporal regulation effect of vascular endothelial growth factor(VEGF)on human fetal choroid vascularization.Methods:The eyeballs of 54human fetuses from the 9th week to the40th ...Purpose:To investigate the spatial and temporal regulation effect of vascular endothelial growth factor(VEGF)on human fetal choroid vascularization.Methods:The eyeballs of 54human fetuses from the 9th week to the40th week due to accidental abortion were studied by immunohistochemically staining for the expression of VEGF and proliferation cell nuclear antigen(PCNA).Results:(1)The distribution of VEGF expression in the retinal pigment epithelium(RPE)decreased with the increase of age,the peak of which was between the 9th and 14th week,(2)PCNAimmunoreactivity was localized within choriocapillaris endothe-lium,THe expression level decreased alone with fetus age,In this period the chori-ocapillaris endothelium kept proliferation,differentiation.canalization and remodelled to form the choroid vessels.(3)Statistically significant correlations were shown between the expression of VEGF in the PRE and that of PCNAin choriocapillaris endothelium(r=0.933,P<0.01).Conclusion:VEGF expression in PRE was positively involved in modulating human fetal choroid vascularization.Eye Science2000;16:11-14.展开更多
目的探讨雌激素对同型半胱氨酸(Hcy)损伤小鼠脑微血管内皮细胞株(bEnd.3)后基质金属蛋白酶(MMP)-9、MMP-2和紧密连接蛋白Occludin表达的影响及其对脑小血管病的保护作用和机制。方法采用Hcy、β-雌二醇分别处理bEnd.3细胞,以细胞增殖及...目的探讨雌激素对同型半胱氨酸(Hcy)损伤小鼠脑微血管内皮细胞株(bEnd.3)后基质金属蛋白酶(MMP)-9、MMP-2和紧密连接蛋白Occludin表达的影响及其对脑小血管病的保护作用和机制。方法采用Hcy、β-雌二醇分别处理bEnd.3细胞,以细胞增殖及毒性检测试剂盒确定最适药物浓度。实验细胞分为对照组(仅加培养液)、Hcy组(200μmol/L的Hcy)、β-雌二醇组(200 nmol/L的β-雌二醇)、联合组(200 nmol/L的β-雌二醇处理30 min后再加入200μmol/L的Hcy),各组均处理6 h。Western blot法检测各组细胞中MMP-9、MMP-2和Occludin表达。结果各组24 h、48 h的MMP-9、MMP-2和Occludin表达比较,差异无统计学意义(P>0.05)。与对照组比较,Hcy组72 h的MMP-9和MMP-2表达明显升高(1.45±0.14 vs 1.00±0.00,1.35±0.15 vs 1.00±0.00,P<0.05),Occludin表达明显降低(0.64±0.05 vs 1.00±0.00,P<0.05)。与Hcy组比较,联合组72 h的MMP-9和MMP-2表达明显降低(0.84±0.19 vs 1.45±0.14,1.01±0.78 vs 1.35±0.15,P<0.05),Occludin表达明显升高(0.91±0.10 vs 0.64±0.05,P<0.05)。结论在Hcy处理的bEnd.3中β-雌二醇能够改善血脑屏障损伤,从而导致MMP-9和MMP-2表达降低,Occludin表达升高。展开更多
文摘Blood vessels constitute a closed pipe system distributed throughout the body,transporting blood from the heart to other organs and delivering metabolic waste products back to the lungs and kidneys.Changes in blood vessels are related to many disorders like stroke,myocardial infarction,aneurysm,and diabetes,which are important causes of death worldwide.Translational research for new appro-aches to disease modeling and effective treatment is needed due to the huge socio-economic burden on healthcare systems.Although mice or rats have been widely used,applying data from animal studies to human-specific vascular physiology and pathology is difficult.The rise of induced pluripotent stem cells(iPSCs)provides a reliable in vitro resource for disease modeling,regenerative medicine,and drug discovery because they carry all human genetic information and have the ability to directionally differentiate into any type of human cells.This review summarizes the latest progress from the establishment of iPSCs,the strategies for differentiating iPSCs into vascular cells,and the in vivo trans-plantation of these vascular derivatives.It also introduces the application of these technologies in disease modeling,drug screening,and regenerative medicine.Additionally,the application of high-tech tools,such as omics analysis and high-throughput sequencing,in this field is reviewed.
基金supported by the national“973”tissue engineering project of China(G1999054300)Shanghai Science and Technology Development Foundation(03DJ14021)
文摘Endothelial cells (TEC3 cells) derived from mouse embryonic stem (ES) cells were used as seed cells to construct blood vessels. Tissue engineered blood vessels were made by seeding 8 × l06 smooth muscle cells (SMCs) obtained from rabbit arteries onto a sheet of nonwoven polyglycolic acid (PGA) fibers, which was used as a biodegradable polymer scaffold. After being cultured in DMEM medium for 7 days in vitro, SMCs grew well on the PGA fibers, and the cell-PGA sheet was then wrapped around a silicon tube, and implanted subcutaneously into nude mice. After 6~8 weeks, the silicon tube was replaced with another silicon tube in smaller diameter, and then the TEC3 cells (endothelial cells differentiated from mouse ES cells) were injected inside the engineered vessel tube as the test group. In the control group only culture medium was injected. Five days later, the engineered vessels were harvested for gross observation, histological and immunohistochemical analysis. The preliminary results demonstrated that the SMC-PGA construct could form a tubular structure in 6~8 weeks and PGA fibers were completely degraded. Histological and immunohistochemical analysis of the newly formed tissue revealed a typical blood vessel structure, including a lining of endothelial cells (ECs) on the lumimal surface and the presence of SMC and collagen in the wall. No EC lining was found in the tubes of control group. Therefore, the ECs differentiated from mouse ES cells can serve as seed cells for endothelium lining in tissue engineered blood vessels.
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The coronavirus disease 2019(COVID-19)infected so far over 250 million people and caused the death of over 5 million worldwide.Aging,diabetes,and cardiovascular diseases,conditions with preexisting impaired endothelial functions predispose to COVID-19.While respiratory epithelium is the main route of virus entry,the endothelial cells(ECs)lining pulmonary blood vessels are also an integral part of lung injury in COVID-19 patients.COVID-19 not only affects the lungs and respiratory system but also gastrointestinal(GI)tract,liver,pancreas,kidneys,heart,brain,and skin.Blood vessels are likely conduits for the virus dissemination to these distant organs.Importantly,ECs are also critical for vascular regeneration during injury/lesions healing and restoration of vascular network.The World Journal of Gastroenterology has published in last two years over 67 outstanding papers on COVID-19 infection with a focus on the GI tract,liver,pancreas,etc.,however,the role of the endothelial and vascular components as major targets for COVID-19-induced tissue injury,spreading to various organs,and injury healing have not been sufficiently emphasized.In the present article,we focus on these subjects and on current treatments including the most recent oral drugs molnupiravir and paxlovid that show a dramatic,significant efficacy in controlling severe COVID-19 infection.
文摘Selenoprotein biosynthesis may not only be affected by the availability of selenium and the transcription rate of pertinent genes but also by the activity of components of the selenocysteine incorporation complex, SelA, B, C, or D. Incorporation of selenocysteine into selenoproteins requires a complex co-translational mechanism guaranteeing the correct recoding of the termination codon TGA as selenocysteine codon. A particular tRNASer(Sec) is enzyrnatically transformed by selenophosphate into tRNAsec which recognizes the UGA codon by means of a specific elongation factor (SelB) and a peculiar mRNA secondary structure. Selenophosphate is formed from selenide and ATP by the SelD gene product, selenophosphate synthase (SelD). To further elucidate the biological role of phospholipid hydroperoxide GPx (PHGPx), we transformed cells with a heterologous (pig) PHGPx gene and/or an additional (human) SelD gene and studied the behaviour of these cells under selenium depletion and repletion. Transfection of the endothelial cell line ECV 304 with either PHGPx cDNA or SelD cDNA did not result in a substantial increase of PHGPx activities, independent of selenium supply. However, cells co-trans fected with both, PHGPx and SelD cDNA, expressed significantly higher PHGPx activlty. This effect was much more pronounced under selenium limiting conditions. The enhanced PHGPx activity correlated with two functional pararneters, increased capability to reduce hydroperoxides and less sensitivity against H2O2-induced cytotoxicity. Thus, the ECV cells, stably transfected with PHGPx and SelD cDNA, provide a model to specifically investigate the role of PHGPx in endothelial cell function
文摘The blood-brain barrier is a unique property of central nervous system blood vessels that protects sensitive central nervous system cells from potentially harmful blood components.The mechanistic basis of this barrier is found at multiple levels,including the adherens and tight junction proteins that tightly bind adjacent endothelial cells and the influence of neighboring pericytes,microglia,and astrocyte endfeet.In addition,extracellular matrix components of the vascular basement membrane play a critical role in establishing and maintaining blood-brain barrier integrity,not only by providing an adhesive substrate for blood-brain barrier cells to adhere to,but also by providing guidance cues that strongly influence vascular cell behavior.The extracellular matrix protein laminin is one of the most abundant components of the basement membrane,and several lines of evidence suggest that it plays a key role in directing blood-brain barrier behavior.In this review,we describe the basic structure of laminin and its receptors,the expression patterns of these molecules in central nervous system blood vessels and how they are altered in disease states,and most importantly,how genetic deletion of different laminin isoforms or their receptors reveals the contribution of these molecules to blood-brain barrier function and integrity.Finally,we discuss some of the important unanswered questions in the field and provide a“to-do”list of some of the critical outstanding experiments.
文摘Purpose:To investigate the spatial and temporal regulation effect of vascular endothelial growth factor(VEGF)on human fetal choroid vascularization.Methods:The eyeballs of 54human fetuses from the 9th week to the40th week due to accidental abortion were studied by immunohistochemically staining for the expression of VEGF and proliferation cell nuclear antigen(PCNA).Results:(1)The distribution of VEGF expression in the retinal pigment epithelium(RPE)decreased with the increase of age,the peak of which was between the 9th and 14th week,(2)PCNAimmunoreactivity was localized within choriocapillaris endothe-lium,THe expression level decreased alone with fetus age,In this period the chori-ocapillaris endothelium kept proliferation,differentiation.canalization and remodelled to form the choroid vessels.(3)Statistically significant correlations were shown between the expression of VEGF in the PRE and that of PCNAin choriocapillaris endothelium(r=0.933,P<0.01).Conclusion:VEGF expression in PRE was positively involved in modulating human fetal choroid vascularization.Eye Science2000;16:11-14.
文摘目的探讨雌激素对同型半胱氨酸(Hcy)损伤小鼠脑微血管内皮细胞株(bEnd.3)后基质金属蛋白酶(MMP)-9、MMP-2和紧密连接蛋白Occludin表达的影响及其对脑小血管病的保护作用和机制。方法采用Hcy、β-雌二醇分别处理bEnd.3细胞,以细胞增殖及毒性检测试剂盒确定最适药物浓度。实验细胞分为对照组(仅加培养液)、Hcy组(200μmol/L的Hcy)、β-雌二醇组(200 nmol/L的β-雌二醇)、联合组(200 nmol/L的β-雌二醇处理30 min后再加入200μmol/L的Hcy),各组均处理6 h。Western blot法检测各组细胞中MMP-9、MMP-2和Occludin表达。结果各组24 h、48 h的MMP-9、MMP-2和Occludin表达比较,差异无统计学意义(P>0.05)。与对照组比较,Hcy组72 h的MMP-9和MMP-2表达明显升高(1.45±0.14 vs 1.00±0.00,1.35±0.15 vs 1.00±0.00,P<0.05),Occludin表达明显降低(0.64±0.05 vs 1.00±0.00,P<0.05)。与Hcy组比较,联合组72 h的MMP-9和MMP-2表达明显降低(0.84±0.19 vs 1.45±0.14,1.01±0.78 vs 1.35±0.15,P<0.05),Occludin表达明显升高(0.91±0.10 vs 0.64±0.05,P<0.05)。结论在Hcy处理的bEnd.3中β-雌二醇能够改善血脑屏障损伤,从而导致MMP-9和MMP-2表达降低,Occludin表达升高。