目的 观察创伤性脑损伤(traumatic brain injury,TBI)大鼠脑组织中水通道蛋白4(aquaporin 4,AQP4)的表达及其时程变化规律,探讨大麻二酚(cannabidiol,CBD)对AQP4的干预作用。 方法 采用改良“Feeney自由落体法”制备大鼠TBI模型,然后随机分为致伤后8 h、1 d、2 d、3 d、5 d、7 d 6个损伤组和6个CBD干预组,另设正常对照组、假手术组。采用免疫荧光单标染色和Western blotting实验观察各组大鼠损伤侧皮质区AQP4蛋白表达情况。 结果 免疫荧光染色结果显示,与假手术组相比,TBI致伤后AQP4阳性表达随时间推移逐渐增高,3 d达到高峰,5 d、7 d有所下降(P<0.05),但仍高于假手术组;CBD干预后1 d、2 d、3 d和5 d的AQP4阳性表达均下降(P<0.05)。Western blotting实验结果显示,与假手术组相比,TBI致伤后8 h、1 d、2 d、3 d的AQP4蛋白表达水平均有所增加(P<0.05);CBD干预后AQP4蛋白表达水平较TBI组均有所下降(P<0.05)。 结论 TBI可引起AQP4表达升高,3 d达到高峰,而CBD能够抑制AQP4过表达,以1 d、2 d、3 d干预效果最为明显。
Abstract
Objective To explore the expression of aquaporin 4 (AQP4) in the brain tissue of rats with traumatic brain injury (TBI) and its time-course change pattern, and to explore the intervention effect of cannabidiol (CBD) on AQP4. Methods The TBI model of rats was prepared by the modified “Feeney free fall method”, and then randomly divided into six injury groups and six CBD intervention groups at 8 h, 1 d, 2 d, 3 d, 5 d, and 7 d after injury. The normal control group (Control group) and the sham operation group (Sham group) were also established. Immunofluorescence single-label staining and Western blotting were used to observe the expression of AQP4 protein in the injured lateral cortical area in each group of rats. Results Immunofluorescence staining result showed that compared with Sham group, AQP4 positive expression gradually increased over time after TBI injury, peaked at 3 d and then decreased at 5 d and 7 d (P<0.05), but still higher than that of Sham group. AQP4 positive expression decreased at 1 d, 2 d, 3 d, and 5 d after CBD intervention (P<0.05). Western blotting result showed that compared with Sham group, AQP4 protein expression increased at 8 h, 1 d, 2 d, and 3 d after TBI injury (P<0.05). AQP4 protein expression decreased after CBD intervention (P<0.05). Conclusions TBI can increase the expression of AQP4, reaching a peak at 3 d. While CBD can inhibit the overexpression of AQP4, the effect of intervention on 1 d, 2 d, and 3 d is the most obvious.
关键词
创伤性脑损伤 /
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大麻二酚 /
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AQP4 /
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大鼠
Key words
Traumatic brain injury /
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Cannabidiol /
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AQP4 /
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Rats
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基金
国家自然科学基金项目(82060241,81960817);云南省科技厅-昆明医科大学应用基础研究联合专项重点项目(202101AY070001-002);云南省创新团队(2019HC014)