基于改良大鼠双侧颈总动脉结扎法建立血管性痴呆模型探讨脑血流量变化规律及对血管新生相关蛋白的影响
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湖南中医药大学针灸推拿与康复学院

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湖南省教育厅科学研究重点项目(22A0253);湖南省中医药科研计划项目(2021233)


Based on modified bilateral carotid artery ligation way to establish vascular dementia rat model and to investigate the changing rules of cerebral blood flow and its effects on angiogenesis related pr
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1.College of Acupuncture & Tuina & Rehabilitation,Hunan Universtiy of Chinese Medicine;2.College of Acupuncture &Tuina & Rehabilitation,Hunan Universtiy of Chinese Medicine

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Research Foundation of Education Bureau of Hunan Province(22A0253);Traditional Chinese Medicine Research Program of Hunan Province(2021233)

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    摘要:

    【摘要】 目的 通过改良大鼠双侧颈总动脉结扎法建立VD大鼠模型观察其脑血流量变化规律及血管新生蛋白表达等变化。 方法 将36只Sprague-Dawley雄性大鼠随机分为假手术组(n=18)和模型组(n=18),假手术组只分离双侧颈总动脉不作结扎,模型组行双侧颈总动脉结扎法制备VD模型,于造模前、造模后14 d采用Morris水迷宫进行行为学测试,激光散斑衬比成像检测脑血流量变化,Western Blotting法检测HIF-1α、VEGF及HO-1蛋白表达,ELISA法检测IL-4、IL-10含量。 结果 造模后14 d,与假手术组比较,模型组逃避潜伏期延长、穿越平台次数减少(P<0.05);造模后2 h、3 d、7 d,模型组较假手术组脑血流量显著降低(P<0.05),造模后第14 d、21 d,假手术组和模型组的脑血流量变化无显著差异(P>0.05);模型组在造模后2 h脑血流量降至最低(P<0.05),此后脑血流量开始恢复,恢复高峰期发生在造模后3-7 d,并于造模后14 d恢复至造模前水平;造模后21 d,模型组海马区HIF-1α、VEGF及HO-1蛋白表达含量较假手术组明显升高(P<0.05),模型组血清中IL-4、IL-10的含量较假手术组明显升高(P<0.05)。 结论 改良大鼠双侧颈总动脉结扎法建立VD模型的脑血流量变化呈现一定时间规律,造模后21 d该模型海马区HIF-1α、VEGF及HO-1的表达显著增加,并伴随IL-4和IL-10含量的升高。

    Abstract:

    【Abstract】 Objective Through the modified bilateral carotid artery ligation way to establish VD rat model to observe the changes of cerebral blood flow and the expression of angiogenic proteins. Methods 36 Sprague-Dawley male rats were randomized into sham group(n=18)and model group(n=18). In the sham group, only the bilateral carotid artery was isolated without ligation, while in the model group, bilateral carotid artery were ligated to establish VD model. Morris water maze behavior test was applied before and 14 days after modeling, the variation of cerebral blood flow was detected by laser speckle contrast imaging. The protein expressions of HIF-1α, VEGF and HO-1 were detected by Western Blotting and the contents of IL-4 and IL-10 were detected by ELISA. Results 14 days after modeling,the escape latency was significantly prolonged and the frequency of crossing the platform had significant decrease for model group as compared with the sham group (P < 0.05). 2 hours,3 days,7 days after modeling, the cerebral blood flow of the model group was significantly lower than that of the sham group (P < 0.05). 14 and 21 days after modeling, there was no significant difference in cerebral blood flow between the sham group and model group (P > 0.05).In the model group, cerebral blood flow decreased to a minimum at 2 hours after modeling (P < 0.05), and then began to recover. The peak of recovery occurred at 3 to 7 days after modeling, and returned to the level before modeling on the 14th day after modeling.At day 21 postoperatively, the expression of HIF-1α, VEGF and HO-1 proteins in hippocampus of the model group were increased remarkably (P < 0.05)and the contents of IL-4 and IL-10 in serum of the model group were significantly rose in comparison with sham group(P < 0.05). Conclusion The variation of cerebral blood flow in VD rats model establish by the modified bilateral carotid artery ligation way showed a certain time rule.At day 21 postoperatively, HIF-1α, VEGF and HO-1 in hippocampus increased significantly, accompanied by increased levels IL-4 and IL-10.

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  • 收稿日期:2023-06-16
  • 最后修改日期:2023-08-03
  • 录用日期:2023-10-22
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