目的 探讨3-甲基腺嘌呤(3-MA)恢复七氟醚导致的幼年大鼠学习记忆能力损伤和神经元凋亡的作用。方法 7日龄SD幼鼠60只,随机数字表法分为七氟醚组、3-MA+七氟醚组、对照组、3-MA组。采用Morris水迷宫评估幼鼠学习记忆能力,采用TUNEL法检测神经元凋亡,比较各组半胱氨酸蛋白酶-3(caspase-3)、海马组织自噬相关蛋白[自噬底物P62、微管相关蛋白1轻链3(LC3)、自噬相关蛋白Beclin1]。结果 第1、2、3、4天七氟醚组逃避潜伏期较对照组显著延长,穿越平台次数较对照组显著减少(P＜0.05);3-MA+七氟醚组逃避潜伏期较七氟醚组显著缩短,穿越平台次数较七氟醚组显著增加(P＜0.05)。麻醉后24 h和行为学测试第1天,高倍镜下对照组细胞形态规整,基本无阳性染色细胞;七氟醚组呈棕黄褐色胞核较对照组明显增多,细胞失去原有的规整形态,细胞核圆形、皱缩、深染,3-MA+七氟醚组阳性染色细胞较七氟醚组明显改善;七氟醚组凋亡率、caspase-3高于对照组,3-MA+七氟醚组凋亡率、caspase-3低于七氟醚组(P＜0.05);与对照组相比,七氟醚组p-PI3K、p-Akt、p-mTOR、P62、LC3、Beclin1更高(P＜0.05),与3-MA+七氟醚组相比则更低(P＜0.05)。结论 3-MA可抑制PI3K/AKT/mTOR的异常激活,逆转海马组织自噬相关蛋白异常,恢复七氟醚导致的幼年大鼠学习记忆能力损伤和神经元凋亡。
Abstract
OBJECTIVE To investigate the role of 3-methyladenine (3-MA) in restoring learning and memory impairment and neuronal apoptosis in young mice due to sevoflurane. METHODS Sixty SD mice of 7 d old were divided into sevoflurane group, 3-MA+sevoflurane group, control group and 3-MA group according to random number table method. Morris water maze was used to evaluate the learning and memory ability of young mice, and the apoptosis of neurons was detected by TUNEL method. The caspase-3, hippocampus autophagy-related proteins [autophagy substrate P62, microtubule-associated protein 1 light chain 3 (LC3), autophagy-related protein Beclin1] were compared in each group. RESULTS On the 1, 2, 3, and 4 d, the evasion incubation period of the sevoflurane group was significantly longer than that of the control group, and the times of crossing the platform were significantly less than those of the control group (P<0.05). The evasion incubation period in the 3-MA+sevoflurane group was significantly shorter than that in the sevoflurane group, and the times of crossing the platform were significantly increased (P<0.05). At 24 h after anesthesia and the first day of the behavioral test, the morphology of the control group cells was regular under high magnification, and there were basically no positive stained cells; the brownish-yellow nuclei in the sevoflurane group were significantly increased compared with the control group, the cells lost their original regular shape, and the nucleus was round, shrunken, and deeply stained. The positive staining cells in the 3-MA+sevoflurane group were significantly improved compared with the sevoflurane group. The apoptosis rate and caspase-3 in the sevoflurane group were higher than those in the control group, the apoptosis rate and caspase-3 in the 3-MA+sevoflurane group were lower than those in the sevoflurane group (P＜0.05). Compared with control group, p-PI3K, p-Akt, p-mTOR, P62, LC3, and Beclin1 in the sevoflurane group were higher (P<0.05), and lower than those in the sevoflurane group (P<0.05). CONCLUSION 3-MA can inhibit the abnormal activation of PI3K/AKT/mTOR, reverse the abnormality of autophagy-related proteins in hippocampus, and restore the impairment of learning and memory and neuronal apoptosis in young mice caused by sevoflurane.
关键词
3-甲基腺嘌呤 /
七氟醚 /
幼年大鼠 /
学习记忆能力 /
神经元凋亡 /
自噬蛋白
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Key words
3-methyladenine /
sevoflurane /
young mice /
learning memory ability /
neuronal apoptosis /
autophagy protein
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中图分类号：
R969.1
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参考文献
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脚注
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基金
浙江省自然科学基金项目资助(ZS5764899)
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