布比卡因脂通过线粒体钙信号轴影响老年麻醉患者认知功能的机制

Mechanisms of bupivacaine liposomes in modulating cognitive function via mitochondrial calcium signaling axis in geriatric anesthesia

摘要

目的：探讨布比卡因（bupivacaine，BPV）脂质通过线粒体钙信号轴对老年麻醉患者术后认知功能障碍（postoperative cognitive dysfunction，POCD）的影响。方法：选择2022年1月—2024年12月在常州市第一人民医院接受全髋关节置换手术的61例患者作为研究对象，分为POCD组（n=15）和非POCD组（n=46）。麻醉方法均为腰麻联合BPV脂质体进行髋关节囊周围神经阻滞。采用ELISA试剂盒检测外周血白细胞线粒体膜呼吸链复合体Ⅰ、Ⅱ、Ⅲ、Ⅳ活性水平。48只18个月大的C57BL/6雄性小鼠随机分为对照组、ASM981组、POCD组和POCD+ASM981组，每组12只。莫里斯水迷宫评估小鼠的空间记忆和学习能力。原代神经元细胞分为空白（Con）组、BPV组、ASM981组、BPV+ASM981组。通过蛋白质印迹法分析AMP依赖的蛋白激酶（adenosine 5′-monophosphate-activated protein kinase，AMPK）和动力相关蛋白1（dynamic related protein 1，Drp1）表达情况，MitoTracker Red标记观察神经元的线粒体。结果：非POCD组老年患者的白细胞线粒体膜呼吸链复合体Ⅰ、Ⅱ、Ⅲ、Ⅳ活性显著高于POCD组（P<0.05）。与POCD组相比，POCD+ASM981组小鼠的逃避潜伏期花费的时间更少，穿过平台的次数增加，在原平台区域停留的时间更长，并且增加了到目标象限的距离（P<0.05）。BPV+ASM981组原代神经元中p-AMPK、mito-Drp1蛋白水平和钙调神经磷酸酶显著低于BPV组（P均&lt; 0.05），p-Ser637-Drp1蛋白水平显著高于BPV组，线粒体分支长度显著长于BPV组（P均&lt; 0.05）。结论：ASM981对BPV诱导的老年小鼠认知功能障碍具有保护作用，作用机制可能是通过抑制AMPK信号通路介导的线粒体分裂，进而改善线粒体功能障碍。

Abstract

Objective: To investigate the effect of bupivacaine (BPV) lipid on cognitive function of elderly anesthesia patients through mitochondrial calcium signal axis modulation. Methods: Sixty-one patients who underwent total hip replacement in our hospital from January 2022 to December 2024 were selected as the research objects and divided into POCD group (n=15) and non-POCD group (n=46). Anesthesia methods were spinal anesthesia combined with bupivacaine liposome for pericapsular nerve block of hip joint. The activity levels of respiratory chain complexes Ⅰ, Ⅱ, Ⅲ, and Ⅳ in mitochondrial membrane of peripheral blood leukocytes were detected by ELISA kit. Forty-eight 18-month-old C57BL/6 male mice were randomly divided into control group, ASM981 group, POCD group and POCD+ASM981 group with 12 mice in each group. Morris water maze was used to evaluate the spatial memory and learning ability of mice. Primary neurons were divided into three groups: blank (Con) group, BPV group, ASM981 group, and BPV+ASM981 group. The expressions of AMP-dependent protein kinase (AMPK) and dynamic related protein 1 (Drp1) were analyzed by western blotting, and MitoTracker Red labeling was used to observe the mitochondria of neurons. Results: The activities of leukocyte mitochondrial membrane respiratory chain complexes Ⅰ, Ⅱ, Ⅲ and Ⅳ in non-POCD group were higher than those in POCD group (P<0.05). Compared with the POCD group, mice in the POCD+ASM981 group spent less time to escape the incubation period, crossed the platform more times, stayed in the original platform area longer, and greater distance traveled in the target quadrant (P<0.05). The levels of p-AMPK, mito-Drp1 protein and calcineurin in primary neurons of the BPV+ASM981 group were lower than those of the BPV group (all P<0.05), while the level of p-Ser637-Drp1 protein was higher than that of the BPV group and the length of mitochondrial branches was longer than that of the BPV group (all P<0.05). Conclusion: ASM981 has protective effect on cognitive dysfunction induced by BPV in aged mice, and the mechanism may be to improve mitochondrial dysfunction by inhibiting mitosis mediated by AMPK signaling pathway.

关键词

布比卡因 / 钙调神经磷酸酶 / 老年小鼠 / 线粒体 / 认知功能

Key words

bupivacaine / calcineurin phosphatase / aged mice / mitochondria / cognitive function

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基金资助

2024年度医院药学高质量发展研究项目专项课题(NIHAZX202415)

常州市高层次人才项目(KY20221373)

常州卫生健康委员会重大项目(ZD202308)