目的 探究銀杏內(nèi)酯B（ginkgolide B，GB）配伍右莰醇（borneol，BO）對腦缺血再灌注損傷大鼠模型的保護作用及機制。方法 采用線栓法建立大鼠大腦中動脈栓塞（middle cerebral artery occlusion，MCAO）模型。將大鼠隨機分為假手術組、模型組、銀杏內(nèi)酯B（10 mg/kg）組、右莰醇（2.5 mg/kg）組、銀杏內(nèi)酯B＋右莰醇（8∶1、4∶1、2∶1）組，拔出線栓后10 min內(nèi)、24 h、48 h各給藥1次，考察大鼠神經(jīng)功能損傷評分、腦梗死面積、腦組織含水量、腦組織病理損傷程度、腦組織炎癥因子、氧化應激及凋亡相關蛋白表達的變化。結果 與假手術組比較，模型組腦梗死面積、神經(jīng)功能評分及腦組織含水量顯著升高（P＜0.001），腦組織病理損傷嚴重；銀杏內(nèi)酯B、右莰醇、銀杏內(nèi)酯B＋右莰醇（4∶1）組的大鼠神經(jīng)功能損傷評分、腦梗死面積、腦組織含水量、神經(jīng)元結構損傷和尼氏小體丟失現(xiàn)象均得到不同程度改善，其中銀杏內(nèi)酯B＋右莰醇（4∶1）組大鼠神經(jīng)行為學評分和腦含水量明顯減少（P＜0.01），且銀杏內(nèi)酯B、右莰醇單獨使用組的作用效果弱于銀杏內(nèi)酯B＋右莰醇（4∶1）組（P＜0.05、0.01）。與假手術組比較，模型組缺血半腦組織中丙二醛（malondialdehyde，MDA）含量顯著增加（P＜0.001），超氧化物歧化酶（superoxide dismutase，SOD）和谷胱甘肽過氧化物酶（glutathione peroxidase，GSH-Px）活性均顯著降低（P＜0.05、0.01），大鼠高遷移率族蛋白B1（high mobility group box 1 protein，HMGB1）、腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）、白細胞介素-6（interleukin-6，IL-6）、白細胞介素-1β（interleukin-1beta，IL-1β）分泌水平顯著升高（P＜0.05、0.001），B淋巴細胞瘤-2基因（B-cell lymphoma-2，Bcl-2）/Bcl-2相關X蛋白（Bcl-2-associated X protein，Bax）的值、核因子E2相關因子2（nuclear factor erythroid 2-related factor 2，Nrf2）、血紅素加氧酶-1（hemeoxygenase-1，HO-1）均顯著降低（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）蛋白表達顯著上調(diào)（P＜0.01）；銀杏內(nèi)酯B＋右莰醇（4∶1）組大鼠腦組織中MDA含量顯著降低（P＜0.01），SOD和GSH-Px活性均顯著升高（P＜0.05、0.01），HMGB1、TNF-α、IL-6和IL-1β分泌水平顯著降低（P＜0.01、0.001），Bcl-2/Bax的值、Nrf2、HO-1均顯著上調(diào)（P＜0.05、0.01），cleaved Caspase-3蛋白表達顯著降低（P＜0.001），且銀杏內(nèi)酯B、右莰醇單獨使用組作用弱于銀杏內(nèi)酯B＋右莰醇（4∶1）組（P＜0.05、0.01、0.001）。結論 銀杏內(nèi)酯B、右莰醇、銀杏內(nèi)酯B＋右莰醇（4∶1）具有神經(jīng)保護作用，且銀杏內(nèi)酯B＋右莰醇（4∶1）的抗腦梗死效應優(yōu)于銀杏內(nèi)酯B、右莰醇單獨使用，其作用機制可能是通過介導Nrf2/HO-1信號通路，進而減輕氧化應激損傷、抑制炎癥反應，減少細胞凋亡從而發(fā)揮的神經(jīng)保護作用。

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

江蘇省基礎研究計劃自然科學基金-前沿引領技術基礎研究專項（BK20232014）