2017年4月5日，《自然》（Nature）在线发表了浙江大学药学院张海涛研究员团队题为《血管紧张素Ⅱ受体的选择性和多样性的结构基础》（structural basis for selectivity and diversity in angiotensin Ⅱ receptors）的论文（http：／...2017年4月5日，《自然》（Nature）在线发表了浙江大学药学院张海涛研究员团队题为《血管紧张素Ⅱ受体的选择性和多样性的结构基础》（structural basis for selectivity and diversity in angiotensin Ⅱ receptors）的论文（http：／／www．nature．com／nature／joumal／vaop／ncurrent／full／nature22035．html）。该研究应用最新的自由电子激光技术（XFEL）和同步辐射，首次解析了AT2R与两种药物先导化合物的复合物晶体三维结构，其中一种是AT2R选择性配体，另一种是AT1R／AT2R双靶点配体，并通过一系列构效关系研究、分子对接、分子动力学和突变实验，系统阐明了不同亚型血管紧张素Ⅱ受体的配体选择性和功能多样性的结构基础，从而为基于结构的AT1R／AT2R靶向药物研究提供了关键信息。展开更多
Objective: To investigate the effect of adenoviral vector-mediated AT2R gene transfection on neointimal hyperplasia after rat carotid artery balloon injury. Methods :AT2R gene was transferred into rat carotid arteries...Objective: To investigate the effect of adenoviral vector-mediated AT2R gene transfection on neointimal hyperplasia after rat carotid artery balloon injury. Methods :AT2R gene was transferred into rat carotid arteries by recombinant adenovirus pAd-AT2R after the establishment of rat carotid balloon injury restenosis model. The arteries were harvested on the 14th day after gene transfer. The efficiency of trans-gene delivery was measured by the expression of adenovirus-encoding green fluorescent protein (GFP) under fluorescent microscope. The expression of AT2R and PCNA (proliferating cell nuclear antigen) was e-valuated by RT-PCR, immunocytochemistry, immunofluorescence staining, confocal microscopy, respectively. The ratio of intimal to medial area (I/M) was quantified with images and determined by an image analysis system. Results: GFP-positive area in adventitia, media and the forming neointima was about 40%. Adenoviral delivery of rat AT2R gene up-regulated AT2R expression in balloon-injured rat carotid and reduced PCNA expression and I/M significantly in neointima(P<0. 01). Double immunofluorescence labeling of AT2R and PCNA also showed that AT2R gene transfer inhibited VSMCs proliferation in neointima. Conclusion: AT2R gene transfer may be a novel promising therapy to limit neointimal hyperplasia.展开更多
目的:血管紧张素Ⅱ 2型受体(AT2R)基因转染的骨髓间充质干细胞(MSC)在四环素可调控系统下作为载体,利用计算机图像分析系统研究新生内膜增生的情况,并探讨AT2R在体可调控表达对大鼠颈动脉损伤后骨桥蛋白(OPN)表达影响。方法:利用球囊损...目的:血管紧张素Ⅱ 2型受体(AT2R)基因转染的骨髓间充质干细胞(MSC)在四环素可调控系统下作为载体,利用计算机图像分析系统研究新生内膜增生的情况,并探讨AT2R在体可调控表达对大鼠颈动脉损伤后骨桥蛋白(OPN)表达影响。方法:利用球囊损伤60只SD大鼠颈动脉,并随机将SD大鼠分为5组,分别为正常组(未行球囊扩张术)、对照组(球囊扩张术后注入PBS)、MSC组(球囊扩张术后注入常规MSC)、MSC转染组(球囊扩张术注入转染AT2R的MSC)、强力霉素(Dox)组(球囊扩张术后注入转染AT2R的MSC,术后当天至处死前三天通过尾静脉注射Dox 100μg/kg/d)。术后14及28天分别处死大鼠取材,光镜下观察血管内膜增生情况,Image pro plus 6.0计算机图像分析系统测量新生内膜面积(I/M),逆转录-多聚酶链反应(RT-PCR)检测AT2R及OPN在大鼠血管标本中的表达变化。结果:大鼠颈动脉AT2R在Dox组的表达显著增高,新的增生内膜面积较其它各损伤组显著降低(P≤0.01),并且OPN的表达显著低于其他各手术组。结论:AT2R基因在体可调控表达受到Dox的有效控制,AT2R基因可能抑制血管损伤后OPN的表达及新生内膜的过度增生。展开更多
文摘2017年4月5日，《自然》（Nature）在线发表了浙江大学药学院张海涛研究员团队题为《血管紧张素Ⅱ受体的选择性和多样性的结构基础》（structural basis for selectivity and diversity in angiotensin Ⅱ receptors）的论文（http：／／www．nature．com／nature／joumal／vaop／ncurrent／full／nature22035．html）。该研究应用最新的自由电子激光技术（XFEL）和同步辐射，首次解析了AT2R与两种药物先导化合物的复合物晶体三维结构，其中一种是AT2R选择性配体，另一种是AT1R／AT2R双靶点配体，并通过一系列构效关系研究、分子对接、分子动力学和突变实验，系统阐明了不同亚型血管紧张素Ⅱ受体的配体选择性和功能多样性的结构基础，从而为基于结构的AT1R／AT2R靶向药物研究提供了关键信息。
文摘Objective: To investigate the effect of adenoviral vector-mediated AT2R gene transfection on neointimal hyperplasia after rat carotid artery balloon injury. Methods :AT2R gene was transferred into rat carotid arteries by recombinant adenovirus pAd-AT2R after the establishment of rat carotid balloon injury restenosis model. The arteries were harvested on the 14th day after gene transfer. The efficiency of trans-gene delivery was measured by the expression of adenovirus-encoding green fluorescent protein (GFP) under fluorescent microscope. The expression of AT2R and PCNA (proliferating cell nuclear antigen) was e-valuated by RT-PCR, immunocytochemistry, immunofluorescence staining, confocal microscopy, respectively. The ratio of intimal to medial area (I/M) was quantified with images and determined by an image analysis system. Results: GFP-positive area in adventitia, media and the forming neointima was about 40%. Adenoviral delivery of rat AT2R gene up-regulated AT2R expression in balloon-injured rat carotid and reduced PCNA expression and I/M significantly in neointima(P<0. 01). Double immunofluorescence labeling of AT2R and PCNA also showed that AT2R gene transfer inhibited VSMCs proliferation in neointima. Conclusion: AT2R gene transfer may be a novel promising therapy to limit neointimal hyperplasia.
文摘目的:血管紧张素Ⅱ 2型受体(AT2R)基因转染的骨髓间充质干细胞(MSC)在四环素可调控系统下作为载体,利用计算机图像分析系统研究新生内膜增生的情况,并探讨AT2R在体可调控表达对大鼠颈动脉损伤后骨桥蛋白(OPN)表达影响。方法:利用球囊损伤60只SD大鼠颈动脉,并随机将SD大鼠分为5组,分别为正常组(未行球囊扩张术)、对照组(球囊扩张术后注入PBS)、MSC组(球囊扩张术后注入常规MSC)、MSC转染组(球囊扩张术注入转染AT2R的MSC)、强力霉素(Dox)组(球囊扩张术后注入转染AT2R的MSC,术后当天至处死前三天通过尾静脉注射Dox 100μg/kg/d)。术后14及28天分别处死大鼠取材,光镜下观察血管内膜增生情况,Image pro plus 6.0计算机图像分析系统测量新生内膜面积(I/M),逆转录-多聚酶链反应(RT-PCR)检测AT2R及OPN在大鼠血管标本中的表达变化。结果:大鼠颈动脉AT2R在Dox组的表达显著增高,新的增生内膜面积较其它各损伤组显著降低(P≤0.01),并且OPN的表达显著低于其他各手术组。结论:AT2R基因在体可调控表达受到Dox的有效控制,AT2R基因可能抑制血管损伤后OPN的表达及新生内膜的过度增生。
