Effect of herpes simplex virus vector, d12CALP, on vasospasm in rabbit common carotid artery with atherosclerotic changes

目的：探讨单纯疱疹病毒（HSV）抑制血管平滑肌细胞增殖对动脉粥样硬化性血管痉挛的影响。方法：在高胆固醇血症家兔颈总动脉周围应用充血硅胶片制备延长性血管痉挛模型。在第0、1和2天（单纯疱疹病毒处理组）将钙钙蛋白启动子驱动ICP4基因和胸苷激酶启动子驱动LacZ基因的HSV载体d12CALP （1×10^9 pfu/ml）注入右颈总动脉。钙钙蛋白是众所周知的在平滑肌细胞中特异性表达的蛋白，因此d12CALP单独在平滑肌细胞中复制并破坏分裂良好的平滑肌细胞。第7天各组兔灌注固定后，用X-gal染色观察HSV处理后的颈总动脉病毒复制分布。形态计量学分析，包括厚度、面积和壁厚的测量。结果：hsv处理第2天组颈动脉平滑肌细胞X-gal染色呈阳性。hsv处理第0、1天各组细胞均未见蓝色染色。形态计量学分析显示，在第2天，无单纯疱疹病毒治疗组的管腔面积明显减少，壁厚明显增加。这些变化在第7天显著持续。然而，与对照组相比，在所有hsv治疗组中，血管没有收缩，而是扩张。其中，hsv治疗第2天组血管舒张程度显著增强（P<0.01）。结论：颈总动脉合并动脉粥样硬化血管痉挛模型第2天血管平滑肌增生。因此，我们认为HSV在此时抑制血管平滑肌细胞增殖可能为治疗血管痉挛提供了一种新的途径。少

Object: The present study was designed to clarify the effect of treatment with herpes simplex virus (HSV) inhibited vascular smooth muscle cell proliferation on vasospasm with atherosclerosis.Methods: We produced a prolonged vasospasm model of Kurosawa and Kusanagi- hypercholesterolemic rabbit common carotid artery by applying a blood-filled silicone sheet around the vessel. A HSV vector, d12CALP, in which the calponin promoter drives ICP4 gene and thymidine kinase promoter drives LacZ gene (1×10^9 pfu/ml) was administered into the right common carotid artery on Days 0, 1 and 2 (HSV-treated groups). Calponin is well-known protein which is specifically expressed in smooth muscle cells, therefore d12CALP replicates in and destroy well dividing smooth muscle cells alone. After perfusion -fixation of each rabbits on Day 7, the common carotid arteries treated with HSV were stained with X-gal to detect the distribution of virus replication. Morphometric analysis including measurements of lum … More inal area and wall thickness was also performed.Results: In HSV-treated on Day 2 group, smooth muscle cells in the carotid artery revealed positive staining with X-gal. However, there was no blue staining in HSV-treated on Days 0 and 1 groups. Morphometric analysis demonstrated a marked reduction in the luminal area as well as increase in the wall thickness on Day 2 in HSV-no treated group. These changes were significantly persistent on Day 7. In all HSV-treated groups, however, there was no constriction of the vessel and it was dilated rather, compared to the control group. Especially, in HSV-treated on Day 2 group, the degree of vasodilatation was significantly strong (P<0.01).Conclusion: In a vasospasm model of common carotid artery with atherosclerosis, vascular smooth muscle proliferation was developed on Day 2. Therefore, we considered that inhibition of the vascular smooth muscle cell proliferation at this time by HSV might provide a novel approach for the treatment of vasospasm. Less