D-松醇缓解脑缺血/再灌注模型大鼠的脑损伤

doi:10.16352/j.issn.1001-6325.2026.03.0359

摘要/Abstract

摘要： 目的探究D-松醇(DP)对脑缺血/再灌注损伤(CI/RI)模型大鼠的治疗作用。方法将大鼠随机分为6组(n=12):假手术组(sham)、模型组(model,改良Longa线栓法建立的CI/RI大鼠模型)、低(L-DP组)、中(M-DP组)和高剂量DP组(H-DP组)(分别灌胃10/20/40 mg/kg的DP),以及H-DP+抑制剂组(inhibitor,灌胃40 mg/kg的DP及腹腔注射30 mg/kg Nrf2抑制剂ML385)。用双盲法进行Zea Longa评分,TTC染色评价脑梗死体积,苏木精-伊红(HE)和尼氏染色观察神经元形态,TUNEL染色标记凋亡细胞。按试剂盒说明检测脑组织SOD活性以及GSH、MDA、TNF-α、IL-1β和IL-6含量。用可见分光光度法检测脑组织匀浆中的Fe²⁺含量。RT-qPCR检测脑组织血红素加氧酶-1(HO-1)、醌氧化还原酶-1(NQO1)、谷胱甘肽过氧化物酶4(GPX4)mRNA水平。Western blot检测脑组织Nrf2(细胞核)、Keap1、HO-1、NQO-1和GPX4蛋白表达水平。结果与假手术组比较,模型组大鼠出现明显脑损伤,神经功能评分、脑梗死体积和TUNEL⁺细胞比例升高(P＜0.05);GSH含量和SOD活性降低(P＜0.05),MDA、TNF-α、IL-1β、IL-6和Fe²⁺水平升高(P＜0.05);GPX4、Nrf2(细胞核)及HO-1和NQO-1蛋白水平降低(P＜0.05),Keap1蛋白水平升高(P＜0.05)。与模型组比较,L/M/H-DP组大鼠脑损伤明显减轻,神经功能评分、脑梗死体积和TUNEL⁺细胞比例均降低(P＜0.05);GSH含量和SOD活性升高(P＜0.05),MDA、TNF-α、IL-1β、IL-6和Fe²⁺水平均降低(P＜0.05);GPX4、Nrf2(细胞核)及HO-1和NQO-1蛋白水平升高(P＜0.05),Keap1蛋白水平降低(P＜0.05)。与H-DP组比较,抑制剂组大鼠脑损伤加重,神经功能评分、脑梗死体积和TUNEL⁺细胞比例均升高(P＜0.05);GSH含量和SOD活性降低(P＜0.05),MDA、TNF-α、IL-1β、IL-6和Fe²⁺水平均升高(P＜0.05);GPX4、Nrf2(细胞核)及HO-1和NQO-1蛋白水平降低(P＜0.05),Keap1蛋白水平升高(P＜0.05)。结论 D-松醇通过抗氧化、抗感染、抗凋亡及抑制铁死亡缓解CI/RI大鼠的脑损伤。

关键词: 脑缺血/再灌注损伤, D-松醇, 核因子E2相关因子2, 抗氧化反应元件, 氧化应激, 铁死亡

Abstract: Objective To investigate the therapeutic effects of D-pinitol (DP) on rats with cerebral ischemia-reperfusion injury (CI/RI). Methods Rats were randomly divided into 6 groups with 12 in each: sham group (sham), model group (CI/RI rat model established by the modified Longa suture method), low (L-DP), medium (M-DP), and high-dose DP group (H-DP) (gavaged with 10, 20, or 40 mg/kg DP, respectively) and H-DP+inhibitor group (inhibitor, gavaged with 40 mg/kg DP and intraperitoneally injected with 30 mg/kg Nrf2 inhibitor ML385). Zea Longa score were evaluated using a double-blind method, cerebral infarction volume was assessed by TTC staining, neuronal morphology was observed via hematoxylin-eosin (HE) and Nissl staining microcopy, apoptotic cells were labeled by TUNEL staining. The activity of SOD and the contents of GSH, MDA, TNF-α, IL-1β and IL-6 in brain tissues were detected by commercial kits, Fe²⁺ content in brain homogenates was measured by visible spectrophotometer. mRNA levels of heme oxygenase-1 (HO-1), NAD (P) H quinone dehydrogenase 1 (NQO1), and glutathione peroxidase 4(GPX4) were analyzed by RT-qPCR. Protein expression of nuclear Nrf2, Keap1, HO-1, NQO1, and GPX4 in brain tissues was detected by Western blot. Results Compared with sham group, the brain injury of rats in the model group showed significant brain injury, and the neurological function score, cerebral infarction volume and the proportion of TUNEL⁺ cells were increased (P＜0.05) while GSH content and SOD activity decreased (P＜0.05). The level of MDA, TNF-α, IL-1β, IL-6 and Fe²⁺ increased (P＜0.05). The protein level of GPX4, Nrf2 (nucleus), HO-1 and NQO-1 decreased (P＜0.05), while the protein level of Keap1 increased(P＜0.05). Compared with model group, the brain injury of rats in L-DP, M-DP and H-DP group was significantly alleviated. The neurological function score, cerebral infarction volume and the proportion of TUNEL⁺ cells were all decreased (P＜0.05). The GSH content and SOD activity increased (P＜0.05), while the levels of MDA, TNF-α, IL-1β, IL-6 and Fe²⁺ all decreased (P＜0.05). The protein levels of GPX4, Nrf2 (nucleus), HO-1 and NQO-1 increased (P＜0.05), while the protein level of Keap1 decreased (P＜0.05). Compared with H-DP group, the brain injury of rats in the inhibitor group was aggravated, and the neurological function score, cerebral infarction volume and the proportion of TUNEL⁺ cells were all increased (P＜0.05). The GSH content and SOD activity decreased (P＜0.05), while the level of MDA, TNF-α, IL-1β, IL-6 and Fe²⁺ all increased(P＜0.05). The protein level of GPX4, Nrf2 (nucleus), HO-1 and NQO-1 decreased (P＜0.05), while the protein level of Keap1 increased (P＜0.05). Conclusions D-pinitol alleviates brain injury in CI/RI rats through anti-oxidation, anti-inflammation, anti-apoptosis effects and inhibition of ferroptosis.

Key words: cerebral ischemia-reperfusion injury, D-pinitol, nuclear factor E2-related factor 2, antioxidant response element, oxidative stress, ferroptosis

中图分类号:

R743.32

徐伯鑫, 山媛, 杨云鹏. D-松醇缓解脑缺血/再灌注模型大鼠的脑损伤[J]. 基础医学与临床, 2026, 46(3): 359-366.

XU Boxin, SHAN Yuan, YANG Yunpeng. D-pinitol alleviates brain injury in rat models with cerebral ischemia-reperfusion[J]. Basic & Clinical Medicine, 2026, 46(3): 359-366.

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