Department of Neurology, The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong, 510632, PR China.
Department of Urology, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shanxi, 710032, PR China.
Biomed Pharmacother. 2019 May;113:108757. doi: 10.1016/j.biopha.2019.108757. Epub 2019 Mar 18.
Previous findings have demonstrated, in a rat model, that chronic cerebral hypoperfusion (CCH) decreases cortical cerebral blood flow (CBF) while Dl-3-n-Butylphthalide (DNB) accelerates the timely recovery of CBF. However, potential biomarkers, therapeutic targets, and underlying mechanisms for these processes are unclear. In this study, a solid-phase antibody microarray for simultaneously detecting multiple proteins was used to search cortex biotargets in CCH compared to a sham control group, and these results were further examined by biological functional analysis. After DNB treatment, western blot and immunostaining were used to verify candidate protein expression. Importantly, we identified seven proteins that may serve as novel biotargets contributing to CCH. The levels of Tie-2, CNTFRα, IL-4, IL-10, ITGAM, MDC, and TROY were uniquely altered in the CCH. The Tie-2 level was significantly decreased and identified in CCH 2 week (W), CCH 4 W and CCH 8 W. In addition, Ang-1 level and Ang-1/Ang-2 ratio were significantly decreased in CCH 2 W and CCH 4 W while Ang-2 level was increased in the CCH, whereas DNB treatment created the inverse effect to some extent. Moreover, the expression of VEGF and CD34 in the earlier stage of CCH and the diameters of bilateral vertebral arteries (VAs), were significantly enlarged by DNB treatment. Together, we found that the Ang-1/Ang-2/Tie-2 signaling axis was altered in the CCH rat cortex, and DNB treatment could timely regulate this angiopoietin/Tie signaling axis to promote neovascularization in early stages.
先前的研究结果表明，在大鼠模型中，慢性大脑低灌注（CCH）会降低皮质脑血流量（CBF），而 Dl-3-n-丁基苯酞（DNB）则加速 CBF 的及时恢复。然而，这些过程的潜在生物标志物、治疗靶点和潜在机制尚不清楚。在这项研究中，使用固相抗体微阵列同时检测多种蛋白质，以寻找 CCH 与假手术对照组相比的皮质生物靶点，并通过生物功能分析进一步研究这些结果。DNB 治疗后，使用 Western blot 和免疫染色来验证候选蛋白的表达。重要的是，我们确定了 7 种可能作为 CCH 新生物靶点的蛋白质。Tie-2、CNTFRα、IL-4、IL-10、ITGAM、MDC 和 TROY 的水平在 CCH 中发生了独特的改变。Tie-2 水平在 CCH 2 周（W）、CCH 4 W 和 CCH 8 W 时显著降低。此外，CCH 2 W 和 CCH 4 W 时 Ang-1 水平和 Ang-1/Ang-2 比值显著降低，而 CCH 时 Ang-2 水平升高，而 DNB 治疗在一定程度上产生了相反的效果。此外，CCH 早期 VEGF 和 CD34 的表达以及双侧椎动脉（VA）的直径均显著增大。综上所述，我们发现 CCH 大鼠皮质中的 Ang-1/Ang-2/Tie-2 信号轴发生改变，DNB 治疗可以及时调节这种血管生成素/Tie 信号轴，促进早期的血管新生。
