目的 探究银杏内酯B（ginkgolide B，GB）配伍右莰醇（borneol，BO）对脑缺血再灌注损伤大鼠模型的保护作用及机制。方法 采用线栓法建立大鼠大脑中动脉栓塞（middle cerebral artery occlusion，MCAO）模型。将大鼠随机分为假手术组、模型组、银杏内酯B（10 mg/kg）组、右莰醇（2.5 mg/kg）组、银杏内酯B＋右莰醇（8∶1、4∶1、2∶1）组，拔出线栓后10 min内、24 h、48 h各给药1次，考察大鼠神经功能损伤评分、脑梗死面积、脑组织含水量、脑组织病理损伤程度、脑组织炎症因子、氧化应激及凋亡相关蛋白表达的变化。结果 与假手术组比较，模型组脑梗死面积、神经功能评分及脑组织含水量显著升高（P＜0.001），脑组织病理损伤严重；银杏内酯B、右莰醇、银杏内酯B＋右莰醇（4∶1）组的大鼠神经功能损伤评分、脑梗死面积、脑组织含水量、神经元结构损伤和尼氏小体丢失现象均得到不同程度改善，其中银杏内酯B＋右莰醇（4∶1）组大鼠神经行为学评分和脑含水量明显减少（P＜0.01），且银杏内酯B、右莰醇单独使用组的作用效果弱于银杏内酯B＋右莰醇（4∶1）组（P＜0.05、0.01）。与假手术组比较，模型组缺血半脑组织中丙二醛（malondialdehyde，MDA）含量显著增加（P＜0.001），超氧化物歧化酶（superoxide dismutase，SOD）和谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）活性均显著降低（P＜0.05、0.01），大鼠高迁移率族蛋白B1（high mobility group box 1 protein，HMGB1）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、白细胞介素-1β（interleukin-1beta，IL-1β）分泌水平显著升高（P＜0.05、0.001），B淋巴细胞瘤-2基因（B-cell lymphoma-2，Bcl-2）/Bcl-2相关X蛋白（Bcl-2-associated X protein，Bax）的值、核因子E2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）、血红素加氧酶-1（hemeoxygenase-1，HO-1）均显著降低（P＜0.05、0.01），剪切型半胱氨酸天冬氨酸蛋白酶3（cleaved cysteine aspartate protease-3，cleaved Caspase-3）蛋白表达显著上调（P＜0.01）；银杏内酯B＋右莰醇（4∶1）组大鼠脑组织中MDA含量显著降低（P＜0.01），SOD和GSH-Px活性均显著升高（P＜0.05、0.01），HMGB1、TNF-α、IL-6和IL-1β分泌水平显著降低（P＜0.01、0.001），Bcl-2/Bax的值、Nrf2、HO-1均显著上调（P＜0.05、0.01），cleaved Caspase-3蛋白表达显著降低（P＜0.001），且银杏内酯B、右莰醇单独使用组作用弱于银杏内酯B＋右莰醇（4∶1）组（P＜0.05、0.01、0.001）。结论 银杏内酯B、右莰醇、银杏内酯B＋右莰醇（4∶1）具有神经保护作用，且银杏内酯B＋右莰醇（4∶1）的抗脑梗死效应优于银杏内酯B、右莰醇单独使用，其作用机制可能是通过介导Nrf2/HO-1信号通路，进而减轻氧化应激损伤、抑制炎症反应，减少细胞凋亡从而发挥的神经保护作用。

Objective To explore the protective effect and mechanism of ginkgolide B (GB) combined with borneol (BO) on cerebral ischemia-reperfusion injury in rats. Methods The model of middle cerebral artery occlusion (MCAO) in rats was established by thread embolism method. Rats were randomly divided into sham group, model group, ginkgolide B (10 mg/kg) group, borneol (2.5 mg/kg) group and ginkgolide B + borneol (8∶1, 4∶1, 2∶1) group, and the drug was given once at 10 min, 24 h and 48 h after the thread plug was pulled out. The changes of neurological impairment score, cerebral infarction area, water content of brain tissue, pathological injury degree of brain tissue, inflammatory factors, oxidative stress and expression of apoptosis-related proteins in brain tissue were investigated. Results Compared with sham group, the area of cerebral infarction, neurological function score and water content of brain tissue in model group were significantly increased (P < 0.001), and the pathological damage of brain tissue was serious. The neurological impairment score, cerebral infarction area, brain tissue water content, neuronal structure damage and Nissl’s corpuscle loss of rats in ginkgolide B, borneol and ginkgolide B + borneol (4∶1) groups were improved to varying degrees, and the neurobehavioral score and brain water content of rats in the ginkgolide B + borneol (4∶1) group were significantly reduced (P < 0.01), and the effect of ginkgolide B and borneol alone was weaker than that of ginkgolide B + borneol (4∶1) (P < 0.05, 0.01). Compared with sham group, the content of Malondialdehyde (MDA) in the ischemic hemisphere tissue of the model group increased significantly (P < 0.001), the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) decreased significantly (P < 0.05, 0.01), and the secretion levels of High mobility group box 1 protein (HMGB1), Tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6) and Interleukin-1beta (IL-1β) increased significantly (P < 0.05, 0.001), and the ratio of B-cell lymphoma-2 (Bcl-2)/Bcl-2-associated X protein (Bax), Nuclear factor erythroid 2-related factor 2 (Nrf2) and Hemeoxygenase-1 (HO-1) decreased significantly (P < 0.05, 0.01), and the expression of cleaved cysteine aspartate protease-3 (cleaved Caspase-3) increased significantly (P < 0.01). In ginkgolide B + borneol (4∶1) group, the content of MDA in brain tissue decreased significantly (P < 0.01), the activities of SOD and GSH-Px increased significantly (P < 0.05, 0.01), and the secretion levels of HMGB1, TNF-α, IL-6 and IL-1β decreased significantly (P < 0.01, 0.001). The ratio of Bcl-2/Bax, Nrf2 and HO-1 were all significantly increased (P < 0.05, 0.01), and the expression of cleaved Caspase-3 was significantly decreased (P < 0.001). Moreover, the effect of ginkgolide B and borneol alone was weaker than that of ginkgolide B + borneol (4∶1) (P < 0.05, 0.01, 0.001). Conclusion ginkgolide B, borneol and ginkgolide B + borneol (4∶1) have neuroprotective effects, and the anti-cerebral infarction effect of ginkgolide B + borneol (4∶1) is better than that of ginkgolide B and borneol alone, and its mechanism may be that it plays a neuroprotective role by mediating Nrf2/HO-1 signaling pathway, thus alleviating oxidative stress injury, inhibiting inflammatory reaction and reducing cell apoptosis.

R285.5

江苏省基础研究计划自然科学基金-前沿引领技术基础研究专项（BK20232014）