毛蕊异黄酮调控Nrf2/HO-1通路对丙泊酚诱导的大鼠神经损伤和认知功能障碍的保护作用研究

doi:10.3760/cma.j.cn441417-20250123-21007

摘要/Abstract

摘要： 目的　探讨毛蕊异黄酮对丙泊酚诱导的大鼠神经损伤和认知功能障碍的影响及作用机制。方法　本研究自2023年6月至2024年1月于陕西省中医医院动物实验室进行。将50只SD雄性大鼠随机分为5组：对照组、丙泊酚组、毛蕊异黄酮低剂量组、毛蕊异黄酮高剂量组、ML385组。后4组腹腔注射丙泊酚100 mg/kg，对照组注射等量生理盐水；毛蕊异黄酮低、高剂量组于腹腔注射丙泊酚后分别灌服50 mg/kg、100 mg/kg毛蕊异黄酮7 d，每天一次；ML385组在每日灌服100 mg/kg毛蕊异黄酮30 min后腹腔注射30 mg/kg核因子E2相关因子2（nuclear factor-E2 related factor-2，Nrf2）抑制剂ML385。通过Morris水迷宫测试评估大鼠学习记忆能力。行为学实验结束后处死各组大鼠，分离双侧海马组织并提取海马神经元细胞。流式细胞术检测细胞凋亡率；Western blot检测海马神经元中天冬氨酸特异性的半胱氨酸蛋白水解酶（cysteinyl aspartate specific proteinase，caspase-3）、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2-related X，Bax）以及Nrf2/血红素氧合酶1（heme oxygenase 1，HO-1）相关通路蛋白表达水平；酶联免疫吸附法（ELISA）检测大鼠海马神经元细胞中炎症因子［白细胞介素（IL）-6和IL-1β］及抗氧化指标［超氧化物歧化酶（superoxide dismutase，SOD）和过氧化氢酶（catalase，CAT）］水平。采用单因素方差分析、LSD法进行统计学分析。结果　Morris水迷宫实验结果表明，毛蕊异黄酮能够改善丙泊酚诱导的大鼠神经损伤和认知功能障碍（均P<0.05）。毛蕊异黄酮能够抑制丙泊酚诱导的大鼠神经凋亡蛋白caspase-3和Bax表达，提高Bcl-2蛋白表达，且呈剂量依赖性（均P<0.05）。毛蕊异黄酮能够抑制丙泊酚诱导的大鼠海马神经元中IL-6和IL-1β水平，提高抗氧化酶SOD和CAT水平（均P<0.05）。毛蕊异黄酮能够提高丙泊酚诱导的大鼠海马神经元中Nrf2、HO-1蛋白表达，激活Nrf2/HO-1信号通路（均P<0.05）。ML385处理可以逆转高剂量毛蕊异黄酮对神经元细胞凋亡、炎症反应的抑制作用，抑制Nrf2、HO-1蛋白表达（均P<0.05）。结论　毛蕊异黄酮通过激活海马神经元中Nrf2/HO-1通路，改善丙泊酚诱导的大鼠神经损伤和认知功能障碍。

关键词: 毛蕊异黄酮, 丙泊酚, 神经损伤, 认知功能障碍, Nrf2/HO-1信号通路, 动物实验

Abstract: Objective　To explore the effects and mechanisms of calycosin on nerve injury and cognitive dysfunction induced by propofol in rats. Methods　This research was conducted from June 2023 to January 2024 at the Animal Laboratory of Shaanxi Provincial Hospital of Traditional Chinese Medicine. Fifty male SD rats were randomly divided into 5 groups: a control group, a propofol group, a low-dose calycosin group, a high-dose calycosin group, and a ML385 group. The last 4 groups were intraperitoneally injected with 100 mg/kg of propofol, while the control group was injected with the same amount of normal saline. The low and high dose groups of calycosin were orally administered 50 mg/kg and 100 mg/kg of calycosin respectively after intraperitoneal injection of propofol, once a day for 7 days. The ML385 group was intraperitoneally injected with 30 mg/kg of nuclear factor-E2 related factor-2 (Nrf2) inhibitor ML385 30 minutes after oral administration of 100 mg/kg of calycosin. The learning and memory abilities of the rats were evaluated through the Morris water maze test. After the behavioral experiments, the rats in each group were sacrificed, and the bilateral hippocampal tissues were isolated and the hippocampal neuron cells were extracted. The apoptosis rate was detected by flow cytometry; the expression levels of cysteinyl aspartate specific proteinase (caspase-3), B-cell lymphoma-2 (Bcl-2), Bcl-2-related X protein (Bax), and Nrf2/heme oxygenase 1 (HO-1) pathway-related proteins in hippocampal neurons were detected by Western blot; the levels of inflammatory factors [interleukin (IL)-6 and IL-1β] and antioxidant indicators [superoxide dismutase (SOD) and catalase (CAT)] in the rat hippocampal neurons were detected by enzyme-linked immunosorbent assay (ELISA). Statistical analysis was conducted using one-way analysis of variance and the LSD method. Results　The results of the Morris water maze experiment showed that calycosin could alleviate the nerve injury and cognitive dysfunction induced by propofol in rats (all P<0.05). Calycosin could inhibit the expressions of the neuroapoptotic proteins caspase-3 and Bax induced by propofol in rats, and increase the expression of Bcl-2 protein in a dose-dependent manner (all P<0.05). Calycosin could inhibit the levels of IL-6 and IL-1β in rat hippocampal neurons induced by propofol, and increase the levels of antioxidant enzymes (SOD and CAT) (all P<0.05). Calycosin could enhance the protein expressions of Nrf2 and HO-1 in rat hippocampal neurons induced by propofol, and activate the Nrf2/HO-1 signaling pathway (all P<0.05). The ML385 treatment could reverse the inhibitory effects of high-dose calycosin on neuron cell apoptosis and inflammatory responses, and it could also inhibit the expressions of Nrf2 and HO-1 proteins (all P<0.05). Conclusion　Calycosin can improve propofol-induced nerve injury and cognitive dysfunction in rats by activating the Nrf2/HO-1 pathway in hippocampal neurons.

Key words: Calycosin, Propofol, Nerve injury, Cognitive dysfunction, Nrf2/HO-1 signaling pathway, Animal experiment

霍燕陈曦李格赵品刘晨白洁. 毛蕊异黄酮调控Nrf2/HO-1通路对丙泊酚诱导的大鼠神经损伤和认知功能障碍的保护作用研究[J]. 国际医药卫生导报, 2025, 31(21): 3551-3557.

Huo Yan, Chen Xi, Li Ge, Zhao Pin, Liu Chen, Bai Jie. Study on the protective effect of calycosin on nerve injury and cognitive dysfunction induced by propofol in rats by regulating the Nrf2/HO-1 pathway[J]. International Medicine and Health Guidance News, 2025, 31(21): 3551-3557.

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