Gaballa M A, Peppel K, Lefkowitz R J, Aguirre M, Dolber P C, Pennock G D, Koch W J, Goldman S
Department of Surgery, Duke University, Durham, NC, USA.
J Mol Cell Cardiol. 1998 May;30(5):1037-45. doi: 10.1006/jmcc.1998.0668.
This study was designed to determine if adenoviral-mediated delivery of a transgene encoding the beta 2-adrenergic receptor (beta 2-AR) to the carotid arterial wall could result in alterations in in vivo vascular function. De-endothelialized rat carotid arteries were infused in vivo with 0.1 mg/ml elastase and adenovirus [6 x 10(9) plaque forming units (PFU)] containing either the marker gene beta-galactosidase (Adeno-beta-gal), DNA encoding the human beta 2-AR (Adeno-beta 2-AR), or no transgene. This low concentration of elastase increased the water permeability (5.2 +/- 0.6 v 1.9 +/- 0.4 x 10(-8) cm/s/mmHg, n = 4, P < 0.0001) without affecting either the vasomotor responsiveness or the morphology of the arterial wall. A transfection efficiency of 73% was achieved with Adeno-beta-gal (n = 3). beta-gal expression was associated with infrequent appearance of T and B lymphocytes, or neutrophil infiltration. Five days after infection with Adeno-beta 2-AR, the total beta-AR density increased six-fold (67.8 +/- 3.4 v 397.0 +/- 155.5 fmol/mg protein, n = 5, P < 0.01); isoproterenol-induced vasorelaxation at transmural pressures from 10-110/mmHg increased (P < 0.01) compared to arteries exposed to control virus (empty adenovirus), n = 4; and isoproterenol-stimulated cAMP production was increased by 65% (n = 5). Thus, adenoviral-mediated delivery of beta 2-ARs into large artery walls results in enhanced beta-AR-mediated vasorelaxation via augmentation in cAMP levels in vascular smooth muscle cells.
本研究旨在确定通过腺病毒介导将编码β2 - 肾上腺素能受体（β2 - AR）的转基因导入颈动脉壁是否会导致体内血管功能的改变。对大鼠去内皮颈动脉在体内注入0.1mg/ml弹性蛋白酶和腺病毒[6×10⁹ 空斑形成单位（PFU）]，腺病毒分别含有标记基因β - 半乳糖苷酶（腺病毒 - β - gal）、编码人β2 - AR的DNA（腺病毒 - β2 - AR）或不含转基因。这种低浓度的弹性蛋白酶增加了水通透性（5.2±0.6对1.9±0.4×10⁻⁸ cm/s/mmHg，n = 4，P < 0.0001），而不影响血管舒缩反应性或动脉壁形态。腺病毒 - β - gal的转染效率达到73%（n = 3）。β - gal表达与T和B淋巴细胞的罕见出现或中性粒细胞浸润相关。用腺病毒 - β2 - AR感染五天后，总β - AR密度增加了六倍（67.8±3.4对397.0±155.5 fmol/mg蛋白，n = 5，P < 0.01）；与暴露于对照病毒（空腺病毒）的动脉相比，在10 - 110/mmHg跨壁压力下异丙肾上腺素诱导的血管舒张增加（P < 0.01），n = 4；异丙肾上腺素刺激的cAMP产生增加了65%（n = 5）。因此，腺病毒介导的将β2 - ARs导入大动脉壁可通过增加血管平滑肌细胞中的cAMP水平增强β - AR介导的血管舒张。
