Zhao Yang, Yang Sheng-Chang, Yu Fu-Yang, Li Wen-Ya, Ji En-Sheng
Department of Physiology, Hebei University of Chinese Medicine, Shijiazhuang 050200, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Mar;36(2):143-147. doi: 10.12047/j.cjap.5904.2020.032.
To investigate the improvement effects of Xiaotan Huayu Liqiao Formula on cognitive impairment in mice exposed to chronic intermittent hypoxia (CIH), and to explore the related mechanisms. Forty-eight male C57 BL/6 mice were randomly divided into four groups as Normoxia, CIH, Formula+CIH and Formula group. Mice were exposed to normoxia in the normoxia and formula group, or intermittent hypoxia in CIH or Formula+CIH group (in the chambers, mice were filled with 100% N to produce FiO of 9% for 1. 5 min. The FiO gradually returned to 21% over the remainder of each cycle. The exposure cycle was repeated every 3 min, 8 h/day for 35 days). Mice were treated with Xiaotan Huayu Liqiao Formula at the dose of 26. 8 g/kg by intragastric administration before CIH exposure. Meanwhile, mice in CIH and normoxia group were given the same volume of normal saline. When the experiment lasts for 26-35 d, Morris water maze was used to detect cognitive dysfunction in mice. At the end of 35 days, Y-maze was performed in each group. After anesthesia, hippocampus was isolated for morphological observation and Western blot ananlysis. Nissl staining and electron microscopy were adopted to assess the neuronal damage in hippocampus, and Western blot was used to detect the levels of PSD-95 and synapsin expression. Compared with normoxia group,the performance of CIH mice was significantly reduced in Morris water maze and Y-maze(<0. 01,<0. 01). Both the number of Nissl staining positive cells and the thickness of the postsynaptic density in hippocampus were significantly reduced. And, the levels of PSD-95 expression in hippocampus was also decreased in the CIH group(<0. 01), however, no significant change of synapsin expression was observed. Compared to CIH group, administration of Xiaotan Huayu Liqiao Formula markedly improved performance of mice in Morris water maze and Y-maze (<0. 01), increased Nissl staining positive cells and the thickness of the postsynaptic density and PSD-95 expression in hippocampus (<0. 01). Xiaotan Huayu Liqiao Formula could alleviate the structural and functional impairment of the postsynaptic dense area, and improved CIH-induced cognitive dysfunction.
探讨消痰化瘀利窍方对慢性间歇性缺氧(CIH)小鼠认知功能障碍的改善作用,并探讨其相关机制。将48只雄性C57 BL/6小鼠随机分为四组,即常氧组、CIH组、方剂+CIH组和方剂组。常氧组和方剂组小鼠暴露于常氧环境,CIH组或方剂+CIH组小鼠暴露于间歇性缺氧环境(在实验舱中,向小鼠充入100%氮气,使吸入氧分数(FiO)为9%,持续1.5分钟。在每个周期的剩余时间内,FiO逐渐恢复到21%。每3分钟重复一次暴露周期,每天8小时,持续35天)。在CIH暴露前,给小鼠灌胃消痰化瘀利窍方,剂量为26.8 g/kg。同时,CIH组和常氧组小鼠给予相同体积的生理盐水。实验持续26 - 35天时,采用Morris水迷宫检测小鼠的认知功能障碍。在35天结束时,对每组小鼠进行Y迷宫实验。麻醉后,分离海马进行形态学观察和蛋白质免疫印迹分析。采用尼氏染色和电子显微镜评估海马神经元损伤,并用蛋白质免疫印迹法检测突触后致密蛋白95(PSD - 95)和突触素表达水平。与常氧组相比,CIH组小鼠在Morris水迷宫和Y迷宫中的表现显著降低(P<0.01,P<0.01)。海马尼氏染色阳性细胞数量和突触后致密物厚度均显著降低。并且,CIH组海马中PSD - 95表达水平也降低(P<0.01),然而,突触素表达未观察到显著变化。与CIH组相比,给予消痰化瘀利窍方显著改善了小鼠在Morris水迷宫和Y迷宫中的表现(P<0.01),增加了海马尼氏染色阳性细胞数量、突触后致密物厚度和PSD - 95表达(P<0.01)。消痰化瘀利窍方可以减轻突触后致密区的结构和功能损伤,改善CIH诱导的认知功能障碍。