银杏内酯注射液及其组分对大鼠缺血性脑卒中模型作用比较及对突触后致密物95表达的影响
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摘要
目的研究银杏内酯注射液(GIs)及其主要组分银杏内酯B(GB)和白果内酯(BB)对急性缺血性脑损伤的改善作用,以及GIs对突触后致密物95(PSD95)蛋白表达的影响。方法①采用Longa法制备大鼠大脑中动脉阻塞(tMCAO)模型,于缺血再灌注1 h后给予GIs(2.5、5.0 mg/kg)、GB(1.25、2.50 mg/kg)、BB(1.25、2.50 mg/kg),每天给药2次,连续给药3 d。于术后24、48、72 h各进行神经功能评分1次;TTC染色法测定脑梗死体积;称大鼠干湿质量,测定脑组织含水量。②大鼠随机分为假手术组、GIs单给药(2.5 mg/kg)组、模型组和GIs治疗(tMCAO+GIs 2.5 mg/kg)组,制备tMCAO模型,再灌1 h后开始ip给药,3 d内每天给药2次。分别于再灌注后24、72 h取脑,Western blotting法检测缺血半影区PSD 95蛋白表达水平。结果①与模型组比较,GIs及其主要组分GB、BB均显著改善tMCAO模型大鼠神经运动功能障碍,减少脑梗死体积,降低脑水肿(P<0.05、0.01);同等剂量下(2.5 mg/kg),GIs治疗效应优于单组份GB、BB。②GIs组给药后胞浆PSD95蛋白水平均较模型组显著升高(P<0.05、0.01)。结论 GIs及其有效组分GB、BB均具有改善tMCAO大鼠急性缺血性损伤的作用,GIs的作用优于GB、BB单独用药,作用机制可能与上调PSD95蛋白表达相关。
Objective To study the effects of Ginkgolides Injection(GIs) and its main components Ginkgolides B(GB) and Baiguolide(BB) on acute ischemic brain injury, and the effects of GIs on the expression of postsynaptic dense substance 95(PSD95).Methods ① A 90 min transcient middle cerebral artery occlusion model(tMCAO) was established using Longa method. Rats were randomly divided into seven groups: tMCAO group, GIs(2.5, 5.0 mg/kg) groups, GB(1.25, 2.5 mg/kg) groups and BB(1.25, 2.5 mg/kg) groups. Rats were ip administered after 1 h of reperfusion, and continuously for three days. Neurological deficit score,infarct size(TTC staining) and brain water content were measured at 72 h after reperfusion. ② Rats were randomly divided into four groups: sham group, sham + GIs groups(2.5 mg/kg), tMCAO groups, and tMCAO + GIs groups(2.5 mg/kg). Rats were ip administered after 1 h of reperfusion, and continuously for three days, twice a day. The PSD 95 protein level in cerebral ischemic penumbra was measured by Western blotting at 24 and 72 h after reperfusion. Results ① Compared with model group, GIs and its main components GB and BB significantly improved neuromotor dysfunction, reduced cerebral infarction volume and brain edema in tMCAO model rats(P < 0.05, 0.01); At the same dose(2.5 mg/kg), the effect of GIs treatment was better than that of single component GB and BB. ② The level of PSD95 protein in GIs group was significantly higher than that in model group(P < 0.05,0.01). Conclusion GIs and its active constituents, GB and BB have therapeutical effect in acute ischemic brain injury induced by tMCAO in rats, and GIs was better than single treatment of GB and BB. The mechanism may be related to the up-regulation of PSD95 protein expression.
Objective To study the effects of Ginkgolides Injection(GIs) and its main components Ginkgolides B(GB) and Baiguolide(BB) on acute ischemic brain injury, and the effects of GIs on the expression of postsynaptic dense substance 95(PSD95).Methods ① A 90 min transcient middle cerebral artery occlusion model(tMCAO) was established using Longa method. Rats were randomly divided into seven groups: tMCAO group, GIs(2.5, 5.0 mg/kg) groups, GB(1.25, 2.5 mg/kg) groups and BB(1.25, 2.5 mg/kg) groups. Rats were ip administered after 1 h of reperfusion, and continuously for three days. Neurological deficit score,infarct size(TTC staining) and brain water content were measured at 72 h after reperfusion. ② Rats were randomly divided into four groups: sham group, sham + GIs groups(2.5 mg/kg), tMCAO groups, and tMCAO + GIs groups(2.5 mg/kg). Rats were ip administered after 1 h of reperfusion, and continuously for three days, twice a day. The PSD 95 protein level in cerebral ischemic penumbra was measured by Western blotting at 24 and 72 h after reperfusion. Results ① Compared with model group, GIs and its main components GB and BB significantly improved neuromotor dysfunction, reduced cerebral infarction volume and brain edema in tMCAO model rats(P < 0.05, 0.01); At the same dose(2.5 mg/kg), the effect of GIs treatment was better than that of single component GB and BB. ② The level of PSD95 protein in GIs group was significantly higher than that in model group(P < 0.05,0.01). Conclusion GIs and its active constituents, GB and BB have therapeutical effect in acute ischemic brain injury induced by tMCAO in rats, and GIs was better than single treatment of GB and BB. The mechanism may be related to the up-regulation of PSD95 protein expression.
引文
[1]黄贱英,孙建宁,梅世昌,等.银杏内酯B对缺血/再灌脑损伤大鼠的保护作用[J].中国药理学通报, 2008, 24(2):269-272.
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[5]邹凯华.白果内酯:银杏叶制剂有效成分研究现状[J].上海医药, 2008; 29(8):365-367.
[6] Janssens D, Michiels C, Delaive E, et al. Protection of hypoxia-induced ATP decrease in endothelial cells by ginkgo biloba extract and bilobalide[J]. Biochem Pharmacol, 1995, 50(7):991-999.
[7]刘科,鄢云彪,丁建花,等.银杏内酯注射液对脑缺血再灌注损伤治疗时间窗及凋亡信号通路的影响[J].药物评价研究, 2018, 41(7):1169-1173
[8]刘颖异,胡玲芹,张映,等. PSD-95与脑缺血的关系及其调控机制研究[J].现代生物医学进展, 2014, 14(9):1796-1800.
[9] Zhou L, Li F, Xu H B, et al. Treatment of cerebral ischemia by disrupting ischemia-induced interaction of nNOS with PSD-95[J]. Nat Med, 2010, 16(12):1439-1443.
[10]俞月萍,徐秋琴,魏尔清,等.急性缺血性卒中的神经保护药物研究进展[J].中国临床药理学与治疗学, 2005,10(8):841-845.
[11] Mehta S L, Manhas N, Raghubir R. Molecular targets in cerebral ischemia for developing novel therapeutics[J].Brain Res Rev, 2007, 54(1):34-66.
[12] Longa E Z, Weinstein P R, Carlson S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989, 20(1):84-91.
[13] Bederson J B, Pitts L H, Tsuji M, et al. Rat middle cerebral artery occlusion:evaluation of the model and development of a neurologic examination[J]. Stroke,1986, 17(3):472-476.
[14] Sheng M, Kim E. The postsynaptic organization of synapses[J]. Cold Spring Harb Perspect Biol, 2011, 3(12):a005678.
[15] Yan B C, Park J H, Ahn J H, et al. Postsynaptic density protein(PSD)-95 expression is markedly decreased in the hippocampal CA1 region after experimental ischemiareperfusion injury[J]. J Neurol Sci, 2013, 330(1-2):111-116.
[16] Gascón S, Sobrado M, Roda J M, et al. Excitotoxicity and focal cerebral ischemia induce truncation of the NR2A and NR2B subunits of the NMDA receptor and cleavage of the scaffolding protein PSD-95[J]. Mol Psychiatry, 2008, 13(1):99-114.
[17] Hardingham G E, Fukunaga Y, Bading H. Extrasynaptic NMDARs oppose synaptic NMDARs by triggering CREB shut-off and cell death pathways[J]. Nat Neurosci, 2002, 5(5):405-414.
[18] Ballarin B, Tymianski M. Discovery and development of NA-1 for the treatment of acute ischemic stroke[J]. Acta Pharmacol Sin, 2018, 39(5):661-668.
[19]于仁娈,金英,郭伟英.银杏叶提取物对大鼠海马缺血再灌注时兴奋性氨基酸释放的影响[J].解放军药学学报, 2003, 19(1):24-26, 62.
[20] Choi D W, Rothman S M. The role of glutamate neurotoxicity in hypoxicischemic neuronal death[J]. Annu Rev Neurosci. 1990, 13:171-182.
[2] Yang Z Z, Li J, Li S X, et al. Effect of ginkgolide B on striatal extracellular amino acids in middle cerebral artery occluded rats[J]. J Ethnopharmacol, 2011, 136(1):117-122.
[3]杨鹏飞,陈卫东.银杏内酯B药理作用研究进展[J].安徽中医学院学报, 2012, 31(5):86-90.
[4]华骏.白果内酯对缺血性脑损伤的神经保护作用及其机制研究[D].南京:南京医科大学, 2017.
[5]邹凯华.白果内酯:银杏叶制剂有效成分研究现状[J].上海医药, 2008; 29(8):365-367.
[6] Janssens D, Michiels C, Delaive E, et al. Protection of hypoxia-induced ATP decrease in endothelial cells by ginkgo biloba extract and bilobalide[J]. Biochem Pharmacol, 1995, 50(7):991-999.
[7]刘科,鄢云彪,丁建花,等.银杏内酯注射液对脑缺血再灌注损伤治疗时间窗及凋亡信号通路的影响[J].药物评价研究, 2018, 41(7):1169-1173
[8]刘颖异,胡玲芹,张映,等. PSD-95与脑缺血的关系及其调控机制研究[J].现代生物医学进展, 2014, 14(9):1796-1800.
[9] Zhou L, Li F, Xu H B, et al. Treatment of cerebral ischemia by disrupting ischemia-induced interaction of nNOS with PSD-95[J]. Nat Med, 2010, 16(12):1439-1443.
[10]俞月萍,徐秋琴,魏尔清,等.急性缺血性卒中的神经保护药物研究进展[J].中国临床药理学与治疗学, 2005,10(8):841-845.
[11] Mehta S L, Manhas N, Raghubir R. Molecular targets in cerebral ischemia for developing novel therapeutics[J].Brain Res Rev, 2007, 54(1):34-66.
[12] Longa E Z, Weinstein P R, Carlson S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989, 20(1):84-91.
[13] Bederson J B, Pitts L H, Tsuji M, et al. Rat middle cerebral artery occlusion:evaluation of the model and development of a neurologic examination[J]. Stroke,1986, 17(3):472-476.
[14] Sheng M, Kim E. The postsynaptic organization of synapses[J]. Cold Spring Harb Perspect Biol, 2011, 3(12):a005678.
[15] Yan B C, Park J H, Ahn J H, et al. Postsynaptic density protein(PSD)-95 expression is markedly decreased in the hippocampal CA1 region after experimental ischemiareperfusion injury[J]. J Neurol Sci, 2013, 330(1-2):111-116.
[16] Gascón S, Sobrado M, Roda J M, et al. Excitotoxicity and focal cerebral ischemia induce truncation of the NR2A and NR2B subunits of the NMDA receptor and cleavage of the scaffolding protein PSD-95[J]. Mol Psychiatry, 2008, 13(1):99-114.
[17] Hardingham G E, Fukunaga Y, Bading H. Extrasynaptic NMDARs oppose synaptic NMDARs by triggering CREB shut-off and cell death pathways[J]. Nat Neurosci, 2002, 5(5):405-414.
[18] Ballarin B, Tymianski M. Discovery and development of NA-1 for the treatment of acute ischemic stroke[J]. Acta Pharmacol Sin, 2018, 39(5):661-668.
[19]于仁娈,金英,郭伟英.银杏叶提取物对大鼠海马缺血再灌注时兴奋性氨基酸释放的影响[J].解放军药学学报, 2003, 19(1):24-26, 62.
[20] Choi D W, Rothman S M. The role of glutamate neurotoxicity in hypoxicischemic neuronal death[J]. Annu Rev Neurosci. 1990, 13:171-182.