高血压大鼠主动脉血管壁细胞增殖和凋亡的原位检测

Detection of thoracic vascular wall cells proliferation and apoptosis in situ in hypertensive rats

目的 观察原位检测高血压大鼠胸主动脉重构过程中血管壁细胞的增殖和凋亡.方法 利用腹主动脉缩窄手术建立肾性高血压大鼠模型,达到稳定高血压状态后取新鲜胸主动脉进行原位增殖和凋亡检测.结果 腹主动脉缩窄术后2～3d大鼠达到稳定的高血压状态.8周后胸主动脉出现血管重构,表现为壁厚增加,内径壁厚比缩小.溴化去氧尿苷（BrdU）渗入法提示血管壁中膜平滑肌细胞增殖水平上调[正常组（10.0±2.1）个/视野,高血压（18.O±3.0）个/视野];末端脱氧核苷酸转移酶脱氧尿苷三磷酸切口末端标记（TUNEL）法提示中膜平滑肌细胞和内膜内皮细胞凋亡水平上升[正常组（14±3）个/视野,高血压（50 ±8）个/视野].使用芝加哥蓝（Chicago蓝）染料能显著降低血管组织自发荧光,使荧光标记的增殖细胞或凋亡细胞清晰可见.结论 通过选择适当的染料和荧光标记,能够在不受血管自发荧光的干扰下原位检测血管壁细胞的增殖和凋亡水平.

Objective To investigate the method of proliferation and apoptosis detection of thoracic aortic remodeling in hypertensive rats in situ.Methods Abdominal aortic coarctation surgery was operated on rats to establish renal hypertension model.Fresh thoracic aorta was dissected to detect proliferation and apoptosis in situ after the model reaching to a steady hypertensive state.Results The rats reached a stable hypertensive state in 2-3 d after the abdominal aortic coarctation surgery.Vascular remodeling of thoracic aorta was observed after 8w,which showed vascular wall thickened and the inner diameter/wall thickness ratio increased.Bromodeoxyuridine （BrdU） infiltration experiment indicated that proliferation of vascular smooth muscle cells in the tunica media elevated [Normal blood pressure group：（10.0 ± 2.1） cells/scopic view,hypertensive group：（18.0± 3.0） cells/scopic view].The method of Terminal deoxyncleotidyl transferase dUTP nick end labeling （TUNEL） showed that apoptosis of the smooth muscle cells in the tunica media and endothelial cells in the tunica intima ascended [Normal blood pressure group：（14 ± 3） cells/scopic view,hypertensive group：（50 ± 8） cells/scopic view].Chicago blue significantly reduced the vascular tissue autofluorescence so that the proliferation or apoptosis of vascular cells with fluorescence labeling were clearly visible.Conclusion By utilizing the appropriate dyes and fluorescent label,it is possible to detect proliferation and apoptosis of vascular vessel in situ without the interference of vascular vessel tissue autofluorescence.