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目的探讨机械通气下吸入一氧化氮(NO)对胎粪吸入综合征(MAS)新生猪脑不同部位胆碱能神经元的保护作用及对星形胶质细胞增殖的影响。方法 建立新生猪胎粪吸入模型,并予间歇正压通气,随机分为胎粪吸入组(n=10)和NO治疗组(n=4),另设正常对照组(n=4)。HE染色计数大脑皮质、海马及基底节神经元细胞,免疫组化法检测皮层、海马及基底节胆碱乙酰基转移酶(ChAT)和胶质纤维酸性蛋白(GFAP)免疫阳性细胞数。结果 胎粪吸入组脑不同部位的神经元细胞和ChAT阳性细胞数均较正常对照组减少[神经元细胞:皮质(29.6±2.3)比(58.2±3.8),海马(27.0±12.7)比(56.0±3.7),基底节(25.0±4.6)比(56.0±4.6);ChAT阳性细胞:皮质(16.7±3.9)比(38.3±9.0),海马(17.5±2.7)比(34.3±11.6),基底节(17.0±1.2)比(35.3±7.1),P均<0.01],GFAP阳性细胞数增多[皮质(54.8±11.0)比(22.0±5.5),海马(54.9±14.7)比(21.0±1.7),基底节(53.8±7.1)比(19.0±4.6),P均<0.001)]。NO治疗组神经元细胞数和ChAT阳性细胞数均多于胎粪吸入组,GFAP阳性细胞数少于胎粪吸入组,P均<0.05。结论 新生猪MAS后吸入NO可减少脑不同部位胆碱能神经元的丢失及星形胶质细胞的增殖,提示吸入NO可能对MAS引起的脑损伤具有保护作用。
Objective To investigate the protective effects of inhalation of nitric oxide (NO) under mechanical ventilation on cholinergic neurons in different parts of newborn piglets with meconium aspiration syndrome (MAS) and on the proliferation of astrocytes. Methods A neonatal model of meconium aspiration was established and intermittently positive pressure ventilation was adopted. The model was randomly divided into meconium inhalation group (n = 10) and NO treatment group (n = 4). The normal control group (n = 4) was also established. The number of neurons in the cerebral cortex, hippocampus and basal ganglia was counted by HE staining. Immunohistochemistry was used to detect the number of immunoreactive cells in the cortex, hippocampus and basal ganglia choline acetyltransferase (ChAT) and glial fibrillary acidic protein (GFAP). Results Compared with the normal control group, the number of neurons and ChAT positive cells in different parts of brain were decreased in meconium aspiration group [the ratio of neurons: cortical (29.2 +/- 2.3) vs (58.2 +/- 3.8), hippocampus (27.0 +/- 12.7) Basal ganglia (25.0 ± 4.6) vs (56.0 ± 4.6); the ratio of ChAT positive cells: cortical (16.7 ± 3.9) vs (38.3 ± 9.0), hippocampus (17.5 ± 2.7) vs (34.3 ± 11.6) (17.0 ± 1.2), (35.3 ± 7.1), P <0.01, respectively. The number of GFAP positive cells increased significantly (54.8 ± 11.0 vs 22.0 ± 5.5, 21.9 ± 14.7 vs 21.0 ± 1.7, Basal ganglia (53.8 ± 7.1) vs (19.0 ± 4.6), P <0.001, respectively]]. The number of neurons and ChAT positive cells in NO treatment group were more than that in meconium aspiration group, and the number of GFAP positive cells was less than that in meconium aspiration group (P <0.05). Conclusion NO inhalation of neonatal pigs can reduce the loss of cholinergic neurons in different parts of the brain and the proliferation of astrocytes, suggesting that NO inhalation may have a protective effect on MAS-induced brain injury.