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目的研究长时间亚麻醉浓度异氟醚暴露对新生大鼠脑星形胶质细胞胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和兴奋性氨基酸转运体2(excitatory amino acid transporter2,EAAT2)表达的影响。方法出生后7 d的Wistar大鼠40只,采用随机数字表法随机分为两组(n=20),异氟醚组(Iso)和对照组(Con),分别暴露于1.1%异氟醚(0.5 MAC)和30%氧气与氮气混合气体中维持6 h。在处理后12、24 h、3和7 d时,两组均分别取乳鼠新鲜脑组织(n=5),冰冻组织切片后,用GFAP抗体和EAAT2抗体进行免疫荧光双标染色,测得额叶皮质区和海马区的星形胶质细胞GFAP、GFAP和EAAT2共染区的荧光强度。结果相比Con组额叶皮质区,Iso组GFAP的荧光强度在处理后12、24 h显著降低(P<0.01),而处理后3、7 d两组差异无统计学意义;海马区比较,Iso组GFAP的荧光强度在处理后12、24 h、3 d相比Con组显著下降(P<0.01),处理后7 d差异无统计学意义。Iso组额叶皮质区和海马区GFAP和EAAT2共染区域的荧光强度在处理后12、24 h、3和7 d均显著降低,与Con组差异有统计学意义(P<0.01)。结论 1.1%异氟醚暴露会暂时性减少新生大鼠额叶皮质区和海马区GFAP表达,延迟星形胶质细胞的骨架发育;而对星形胶质细胞EAAT2表达抑制是持续性的,这可能是异氟醚对新生大鼠产生脑毒性的机制之一。
Objective To study the effects of prolonged exposure to isoflurane at sub-anesthetic concentrations on the expression of glial fibrillary acidic protein (GFAP) and excitatory amino acid transporter 2 (EAAT2) in neonatal rat astrocytes Impact. Methods Forty Wistar rats, 7 days after birth, were randomized into two groups (n = 20), Iso and control groups (Con) and were exposed to 1.1% isoflurane (0.5 MAC) and 30% oxygen and nitrogen gas mixture for 6 h. At 12, 24 h, 3 and 7 d after treatment, fresh brain tissue of neonatal rats was obtained in each group (n = 5). Frozen sections were harvested and immunofluorescence double-labeled with GFAP antibody and EAAT2 antibody Fluorescence intensity of GFAP, GFAP and EAAT2 co-staining area of astrocytes in frontal cortex and hippocampus. Results Compared with Con group, the fluorescence intensity of GFAP in Iso group was significantly decreased at 12 and 24 h after treatment (P <0.01), while there was no significant difference between the two groups at 3 and 7 days after treatment (P <0.05) The fluorescence intensity of GFAP in Iso group was significantly lower than that in Con group at 12, 24 and 3 days after treatment (P <0.01), but there was no significant difference at 7 days after treatment. The fluorescence intensity of GFAP and EAAT2 in frontal cortex and hippocampus of Iso group were significantly decreased at 12, 24, 3 and 7 days after treatment, which was significantly different from Con group (P <0.01). Conclusion 1.1% isoflurane exposure can temporarily reduce the expression of GFAP in the frontal cortex and hippocampus of neonatal rats, delay the skeletal development of astrocytes, and inhibit the expression of EAAT2 in astrocytes continuously It may be one of the mechanisms of isoflurane on brain toxicity in neonatal rats.