高压氧对成年缺氧缺血性脑病大鼠白质损伤的作用机制

doi:10.20047/j.issn1673-7210.2023.17.05

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摘要目的探讨高压氧（HBO）对成年缺氧缺血性脑病大鼠白质损伤的作用机制。方法 48只300～350 g的10周龄、清洁级雄性SD大鼠采用完全随机化方法分为四组：对照组（不予任何处理）、假手术组（仅暴露双侧颈动脉后直接缝合皮肤）、模型组（结扎双侧颈动脉2周后将大鼠置于通8%O2、92%N2混合气的密闭箱中3 h）、HBO组（在模型组基础上立即进行HBO处理），每组12只。分别于造模前和第4周通过水迷宫评价大鼠学习记忆功能；Fast Blue染色观察大鼠海马区白质髓鞘损伤；免疫荧光法测定大鼠海马区离子钙结合衔接分子1（Iba-1）、胶质纤维酸性蛋白（GFAP）表达；Western blot法分析海马区髓鞘碱性蛋白（MBP）、白细胞介素（IL）-1β、IL-6及肿瘤坏死因子（TNF-α）的蛋白表达。结果假手术组与对照组平均潜伏期及平台穿越次数比较，差异无统计学意义（P＞0.05）。模型组平均潜伏期长于对照组，平台穿越次数低于对照组（P＜0.05）。HBO组平均潜伏期短于模型组，平台穿越次数高于模型组（P＜0.05）。假手术组与对照组比较，大鼠海马区染色程度基本相同，髓鞘无较大区别。模型组与对照组比较，大鼠海马区的染色程度减弱，髓鞘减少，部分区域出现空泡。HBO组与模型组比较，大鼠海马区染色程度好转，髓鞘增加及区域空泡明显减少。假手术组与对照组海马区GFAP阳性的活化星形胶质细胞和Iba-1阳性的激活态小胶质细胞数量比较，差异无统计学意义（P＞0.05）。模型组海马区GFAP阳性的活化星形胶质细胞及Iba-1阳性的激活态小胶质细胞数量高于对照组（P＜0.05）。HBO组海马区GFAP阳性的活化星形胶质细胞及Iba-1阳性的激活态小胶质细胞数量低于模型组（P＜0.05）。假手术组与对照组MBP、IL-1β、IL-6和TNF-α表达比较，差异无统计学意义（P＞0.05）。模型组脑内MBP蛋白表达低于对照组，IL-1β、IL-6和TNF-α表达高于对照组（P＜0.05）。HBO组脑内MBP蛋白表达高于模型组，IL-1β、IL-6和TNF-α蛋白表达水平低于模型组（P＜0.05）。结论高压氧可能通过抑制海马区小胶质细胞和星形胶质细胞活化，减少炎症因子释放，减轻髓鞘损伤和促进髓鞘再生，从而改善成年缺氧缺血性脑病大鼠认知损害。

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	薛瑞君1　　李文平2　　黄蔚喆3　　潘树义1　　郭大志1

关键词 ：缺氧缺血性脑病, 高压氧, 白质损伤, 髓鞘, 炎症

Abstract：Objective To investigate the action mechanism of hyperbaric oxygen (HBO) on white matter damage in adult rats with hypoxic-ischemic encephalopathy. Methods Forty-eight ten- week-old clean grade male SD rats aged 300-350 g were divided into four groups by completely randomized method: control group (no treatment), sham operation group (the skin was sutured directly after only bilateral carotid arteries were exposed), model group (two weeks after bilateral carotid arteries were ligation, rats were placed in a sealed chamber with 8%O2 and 92%N2 mixture for three hours), HBO group (HBO treatment was immediately given on the basis of model group), 12 rats in each group. The learning and memory function was evaluated by water maze before modeling and in the 4th week respectively. Fast Blue staining was used to observe the white matter myelin sheath damage in rat hippocampus. The expressions of ionized calcium binding adapter molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP) in the hippocampus were determined by immunofluorescence method. The expressions of myelin basic protein (MBP), interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) in hippocampus were analyzed by Western blot. Results There were no significant differences in the average incubation period and the number of platform crossing between the sham operation group and the control group (P＞0.05). The average incubation period of the model group was longer than that of the control group, and the number of platform crossing was lower than that of the control group (P＜0.05). The average incubation period of the HBO group was shorter than that of model group, and the number of platform crossing was higher than that of the model group (P＜0.05). The sham operation group was compared with the control group, the hippocampal staining degree of rats was basically the same, and there was no significant difference in myelin sheath. Compared with the control group, the hippocampal staining of rats was weakened, the myelin sheath was reduced, and vacuoles were appeared in some areas in the model group. Compared with the model group, hippocampal staining degree of rats was improved, myelin sheath was increased and regional vacuoles were decreased significantly in the HBO group. There was no significant difference in the number of GFAP-positive activated astrocytes and Iba-1 positive activated microglia in the hippocampus between the sham operation group and the control group (P＞0.05). The number of GFAP-positive activated astrocytes and Iba-1 positive activated microglia in the hippocampus of the model group was higher than that of the control group (P＜0.05). The number of GFAP-positive activated astrocytes and Iba-1 positive activated microglia in hippocampus of the HBO group was lower than that of the model group (P＜0.05). There were no significant differences in the expressions of MBP, IL-1β, IL-6, and TNF-α between the sham operation group and the control group (P＞0.05). The expression of MBP protein in the brain of the model group was lower than that of the control group, and the expressions of IL-1β, IL-6, and TNF-α were higher than those of the control group (P＜0.05). The expression of MBP protein in the HBO group was higher than that in the model group, but the expression levels of IL-1β, IL-6, and TNF-α in the HBO group were lower than that in the model group (P＜0.05). Conclusion HBO may reduce the activation of microglias and astrocytes in the hippocampus, inhibit the release of inflammatory factors, relieve myelin sheath damage, and promote myelin sheath regeneration, thereby improving the cognitive impairment of adult rats with hypoxic-ischemic encephalopathy.

Key words： Hypoxic-ischemic encephalopathy Hyperbaric oxygen White matter damage Myelin sheath Inflammation

基金资助:首都卫生发展科研专项项目（首发2018-4-5111）；
北京市自然科学基金资助项目（7222184）；
河南省医学科技攻关计划项目（LHGJ20200738）。

通讯作者: 潘树义（1963.3-），男，医学博士，主任医师，硕士生导师；研究方向：高压氧在神经保护治疗中的作用与机制。郭大志（1983.11-），男，医学博士，副主任医师，副教授；研究方向：高压氧在脑损伤中的作用和机制。

引用本文:

薛瑞君1　　李文平2　　黄蔚喆3　　潘树义1　　郭大志1. 高压氧对成年缺氧缺血性脑病大鼠白质损伤的作用机制[J]. 中国医药导报, 2023, 20(17): 26-30.
XUE Ruijun1 LI Wenping2 HUANG Weizhe3 PAN Shuyi1 GUO Dazhi1. Action mechanism of hyperbaric oxygen on white matter damage in adult rats with hypoxic-ischemic encephalopathy. 中国医药导报, 2023, 20(17): 26-30.

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