Wu X, Zhang J, Huang Q, Yang P, Chen J, Liu J
Changhai Hospital Department of Neurosurgery Shanghai China.
PLA 455 Hospital Department of Neurosurgery Shanghai China.
Cell Mol Biol (Noisy-le-grand). 2015 Nov 8;61(7):33-9.
We investigated expression of Kruppel—like factor 2 (KLF2) and its correlation with basilar artery blood flow rate in the hemodynamically induced aneurysm model built by different methods. New Zealand rabbits were randomly divided into sham—operated group, unilateral ligation of common carotid artery (CCA) group (UL group) and bilateral ligation of CCA group (BL group). Rabbits were cervix—cut to expose the arteries without ligation (sham group), with right—side ligation (UL group) and bilateral ligation (BL group), respectively. Skull Doppler ultrasound was used to measure basilar artery blood flow rate in each group at week 1, 2, 3, or 4 separately (n=6 for each time point). The animals were killed after the measurements. At each time point, 6 basilar artery bifurcates from each group were collected and sent for staining (HE, EVG, Masson and KLF2 immunohistochemistry staining), while another 6 basilar artery bifurcates were processed with KLF2 Western blotting. Results showed that the average blood flow rate did not change significantly among the 4 time points in the Sham group, but it was insignificantly smaller compared to the UL group. The average blood flow rate in the BL group was significantly higher than that in the other two groups. Pathological tests showed that according to the aneurysm evaluation criteria, the basilar arteries in the Sham group had smooth tip lumina, complete endothelial cells, complete internal elastic membranes, but no fracture, thinning or aneurysm formation. Only 1 of 24 rabbits in the UL group had slight bulges in the tip of basilar artery, though not very severe. Twelve rabbits in the BL group had aneurysmal bulges, significantly different from the other two groups. KLF2 protein expression was not changed significantly with time in the Sham group, but increased slightly with time in the UL group. KLF2 protein expression in the BL group increased significantly only after 1 week and then maintained a high level, significantly different from the other two groups. KLF2 protein expression was positively correlated with blood flow rate. In conclusion, the hemodynamic change was most significant after bilateral ligation, which was most suitable for building animal models for aneurysm research. aaaAnd KLF2 expression change was consistent with blood flow rate variation, which showed positive correlation, indicating KLF2 expression was regulated by hemodynamic changes.
我们研究了在通过不同方法构建的血流动力学诱导动脉瘤模型中，Kruppel样因子2（KLF2）的表达及其与基底动脉血流速度的相关性。将新西兰兔随机分为假手术组、单侧颈总动脉结扎（CCA）组（UL组）和双侧颈总动脉结扎（BL组）。分别对兔进行颈部切开以暴露未结扎的动脉（假手术组）、右侧结扎（UL组）和双侧结扎（BL组）。在第1、2、3或4周分别使用颅骨多普勒超声测量每组的基底动脉血流速度（每个时间点n = 6）。测量后处死动物。在每个时间点，从每组收集6个基底动脉分叉处并送去进行染色（苏木精-伊红染色、弹性纤维染色、马松染色和KLF2免疫组织化学染色），而另外6个基底动脉分叉处用KLF2蛋白质印迹法处理。结果显示，假手术组4个时间点的平均血流速度无明显变化，但与UL组相比略有降低。BL组的平均血流速度明显高于其他两组。病理检查表明，根据动脉瘤评估标准，假手术组基底动脉尖端管腔光滑，内皮细胞完整，内弹性膜完整，但无断裂、变薄或动脉瘤形成。UL组24只兔中只有1只基底动脉尖端有轻微凸起，程度不太严重。BL组12只兔有动脉瘤样凸起，与其他两组有显著差异。假手术组KLF2蛋白表达随时间无明显变化，但UL组随时间略有增加。BL组KLF2蛋白表达仅在1周后显著增加，然后维持在高水平，与其他两组有显著差异。KLF2蛋白表达与血流速度呈正相关。总之，双侧结扎后血流动力学变化最为显著，最适合构建动脉瘤研究的动物模型。并且KLF2表达变化与血流速度变化一致，呈正相关，表明KLF2表达受血流动力学变化调节。
