银杏提取物缓解丙泊酚麻醉大鼠海马组织损伤的机制研究
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摘要
目的:探究银杏提取物(GBE)对麻醉大鼠海马环腺苷酸反应元件结合蛋白-1(CREB1)、脑源性神经营养因子(BD-NF)表达的影响。方法:将大鼠随机分为正常对照组、模型组、银杏提取物低(GBE-L,40 mg·kg~(-1))、中(GBE-M,60 mg·kg~(-1))、高(GBE-H,80 mg·kg~(-1))剂量组、盐酸多奈哌齐(donepezil,1.67 mg·kg~(-1))组,模型组经腹腔注射100 mg·kg~(-1)丙泊酚麻醉,GBE各组和Donepezil组在麻醉前腹腔注射相应药物,正常对照组经腹腔注射生理盐水,麻醉结束后测定大鼠舒张压(DAP)、收缩压(SAP)、心率(HR);麻醉结束后第2天行水迷宫实验,大鼠断颈处死后,HE染色观察海马组织的变化,免疫印迹(WB)和免疫组化法检测CREB1、p-CREB1、BDNF、caspase-3、Bal-2蛋白表达。结果:与正常对照组相比,模型组SAP、DAP、逃避潜伏期、caspase-3蛋白表达均显著升高,穿越原平台次数、Bal-2、BDNF、p-CREB1蛋白以及蛋白阳性率表达均显著降低(P<0.05)。与模型组相比,GBE各剂量组及Donepezil组SAP、DAP、逃避潜伏期、caspase-3蛋白表达显著降低,穿越原平台次数、Bal-2、BDNF、p-CREB1蛋白以及蛋白阳性率表达显著升高(P<0.05)。正常对照组海马神经元细胞结构完整,形态正常,细胞紧密排列染色质分布均匀;模型组细胞形态不规则,排列紊乱,细胞膜破碎,细胞融合,界限模糊;经GBE干预后神经元细胞逐渐恢复正常。结论:GBE能够缓解麻醉药物对大鼠海马组织的损伤,可能与上调BDNF表达以及激活CREB1磷酸化有关。
Objective: To explore the influences of ginkgo biloba extract( GBE) on the expressions of cAMP response element binding protein-1( CREB1) and brain derived neurotrophic factor( BDNF) in the hippocampal tissue of anesthetized rats. Methods:The rats were randomly divided into the normal control group,the model group,GBE low( GBE-L,40 mg·kg~(-1)),medium( GBEM,60 mg·kg~(-1)) and high( GBE-H,80 mg·kg~(-1)) dose groups,and donepezil group( 1. 67 mg·kg~(-1)). The model group was anesthetized by intraperitoneal injection of 100 mg·kg~(-1) propofol,GBE groups and donepezil group were given intraperitoneal injection of corresponding drug before anesthesia,and the normal control group was given intraperitoneal injection of normal saline. The diastolic blood pressure( DAP),systolic blood pressure( SAP) and heart rate( HR) were measured after anesthesia. The water maze test was performed on the second day after anesthesia. The rats were sacrified by breaking necks,and then HE staining was used to observe the changes of hippocampal tissues,and the expressions of CREB1,p-CREB1,BDNF,caspase-3 and Bal-2 were detected by Western blot( WB) and immunohistochemistry. Results: Compared with those in the blank control group,SAP,DAP,escape latency and caspase-3 protein expressions in the model group were increased,and the number of passing through the original platform,Bal-2,p-CREB1,BDNF protein and protein positive expression rate were all decreased( P < 0. 05). Compared with those in the model group,SAP,DAP,escape latency and caspase-3 protein expressions in all GBE groups and donepezil group were decreased,and the number of passing through the original platform,Bal-2,p-CREB1,BDNF protein and protein positive expression rate were all increased( P <0. 05). The hippocampal neurons in the blank control group had complete structure,normal morphology and well-distributed chromatin,the cells in the model group had irregular morphology,disordered arrangement,broken cell membranes,cell fusion and blurred boundaries,and after GBE intervention,the neurons gradually returned to normal. Conclusion: GBE can alleviate the anesthetic injury of hippocampal tissue in rats,which may be related to the up-regulation of BDNF expression and the activation of CREB1 phosphorylation.
Objective: To explore the influences of ginkgo biloba extract( GBE) on the expressions of cAMP response element binding protein-1( CREB1) and brain derived neurotrophic factor( BDNF) in the hippocampal tissue of anesthetized rats. Methods:The rats were randomly divided into the normal control group,the model group,GBE low( GBE-L,40 mg·kg~(-1)),medium( GBEM,60 mg·kg~(-1)) and high( GBE-H,80 mg·kg~(-1)) dose groups,and donepezil group( 1. 67 mg·kg~(-1)). The model group was anesthetized by intraperitoneal injection of 100 mg·kg~(-1) propofol,GBE groups and donepezil group were given intraperitoneal injection of corresponding drug before anesthesia,and the normal control group was given intraperitoneal injection of normal saline. The diastolic blood pressure( DAP),systolic blood pressure( SAP) and heart rate( HR) were measured after anesthesia. The water maze test was performed on the second day after anesthesia. The rats were sacrified by breaking necks,and then HE staining was used to observe the changes of hippocampal tissues,and the expressions of CREB1,p-CREB1,BDNF,caspase-3 and Bal-2 were detected by Western blot( WB) and immunohistochemistry. Results: Compared with those in the blank control group,SAP,DAP,escape latency and caspase-3 protein expressions in the model group were increased,and the number of passing through the original platform,Bal-2,p-CREB1,BDNF protein and protein positive expression rate were all decreased( P < 0. 05). Compared with those in the model group,SAP,DAP,escape latency and caspase-3 protein expressions in all GBE groups and donepezil group were decreased,and the number of passing through the original platform,Bal-2,p-CREB1,BDNF protein and protein positive expression rate were all increased( P <0. 05). The hippocampal neurons in the blank control group had complete structure,normal morphology and well-distributed chromatin,the cells in the model group had irregular morphology,disordered arrangement,broken cell membranes,cell fusion and blurred boundaries,and after GBE intervention,the neurons gradually returned to normal. Conclusion: GBE can alleviate the anesthetic injury of hippocampal tissue in rats,which may be related to the up-regulation of BDNF expression and the activation of CREB1 phosphorylation.
引文
1邓小园,陈博,刘红亮,等.TNF-α在丙泊酚诱发的神经元凋亡及认知功能障碍中的作用[J].中国药理学通报,2016,32(7):945-949
2张蕴,林如辉,李钻芳,等.电针神庭、百会对脑缺血再灌注大鼠学习记忆能力和海马CA1区环磷酸腺苷效应元件结合蛋白及其磷酸化水平的影响[J].中国康复理论与实践,2016,22(11):1241-1245
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9饶婉宜,鲁惠顺.异氟烷诱导术后认知功能障碍的机制及预防的研究进展[J].中国药理学与毒理学杂志,2016,30(2):178-182
10李世勇,陈欣,陈晔凌,等.氯化锂预先给药对异氟醚麻醉诱发老龄大鼠认知功能障碍及海马炎性反应的影响[J].中华麻醉学杂志,2013,33(1):24-27
11胡茂林,宗小芬,梁炜,等.抑郁症模型大鼠海马区磷酸化环磷腺苷反应元件结合蛋白1表达水平及神经元和突触超微结构的研究[J].中华精神科杂志,2015,48(2):92-97
12 Zhou L,Zhang Y,Liu RY,et al.Rescue of impaired long-term facilitation at sensorimotor synapses of Aplysia following siRNA knockdown of CREB1[J].J Neurosci,2015,35(4):1617-1726
13 Jr MG,Sarginson JE,Ryan HS,et al.BDNF and CREB1 genetic variants interact to affect antidepressant treatment outcomes in geriatric depression[J].Pharmacogenet Genomics,2013,23(6):301-313
14彭生,杨建平,张晓庆,等.磷酸二酯酶-4抑制剂Ro20-1724对氯胺酮麻醉后学习记忆及大鼠海马c AMP/PKA-CREB-BDNF信号通路的影响[J].中华行为医学与脑科学杂志,2014,23(6):528-530
15孙东,杨静娴,张囡,等.牛蒡苷元干预CREB/BDNF信号通路保护Aβ25-35损伤神经元的研究[J].中华中医药学刊,2014,19(2):288-290
16 Zhang JC,Yao W,Ren Q,et al.Depression-like phenotype bydeletion ofα7 nicotinic acetylcholine receptor:Role of BDNF-TrkB in nucleus accumbens[J].Sci Rep,2016,6(13):36705-36712
17 Liu ME,Huang CC,Chen MH,et al.Effect of the BDNF Val66Met polymorphism on regional gray matter volumes and cognitive function in the Chinese population[J].Neuromolecular Med,2014,16(1):127-136
18 Hajirezaii B,Barari A,Daloii AA.Consumption of Ginkgo biloba extract and swimming training on NMDA and BDNF plasma[J].Development,2015,141(10):2108-2118
19李金凤,徐桂梅,张辰子,等.银杏叶提取物对MPTP诱导帕金森小鼠保护作用的研究[J].中国比较医学杂志,2016,26(1):46-53
20傅亚,谈宗华,胡承波,等.银杏叶提取物对中枢神经系统保护作用的研究进展[J].中国药房,2013,14(23):2196-2199
21鄢云彪,华骏,兰新新,等.银杏内酯注射液对脑缺血再灌注大鼠运动认知功能的影响[J].中药药理与临床,2018,34(1):138-140
22 Jiang H,Zhao PJ,Su D,et al.Paris saponin I induces apoptosis via increasing the Bax/Bcl-2 ratio and caspase-3 expression in gefitinibresistant non-small cell lung cancer in vitro and in vivo[J].Mol Med Rep,2014,9(6):2265-2271
2张蕴,林如辉,李钻芳,等.电针神庭、百会对脑缺血再灌注大鼠学习记忆能力和海马CA1区环磷酸腺苷效应元件结合蛋白及其磷酸化水平的影响[J].中国康复理论与实践,2016,22(11):1241-1245
3付燕,谢攀,李雪,等.长期有氧运动对大鼠脑衰老过程中学习记忆与海马BDNF表达的影响[J].中国运动医学杂志,2015,34(8):750-756
4 Ghodratijaldbakhan S,Khalili MK,Vafaei AA,et al.Effects of different intensities levels of treadmill exercise on cognitive functions and BDNF levels in prefrontal cortex of morphine dependent rats[J].Koomesh,2016,17(3):669-676
5许佩龙,刘健,李雪飞,等.丙泊酚麻醉对新生期大鼠海马BD-NF及Trk-B的影响[J].中国药理学通报,2012,28(6):858-861
6胡海燕,雷磊,陈翔,等.清心开窍方挥发油对AD大鼠大脑皮层及海马区GFAP及Caspase-3表达的影响[J].中国中西医结合杂志,2013,33(7):927-932
7 Bercker S,Bert B,Bittigau P,et al.Neurodegeneration in newborn rats following propofol and sevoflurane anesthesia[J].Neurotox Res,2009,16(2):140-146
8罗涛,刘志刚,刘敏,等.中枢组胺能神经介导异氟烷麻醉后认知功能障碍[J].神经损伤与功能重建,2012,7(6):407-410
9饶婉宜,鲁惠顺.异氟烷诱导术后认知功能障碍的机制及预防的研究进展[J].中国药理学与毒理学杂志,2016,30(2):178-182
10李世勇,陈欣,陈晔凌,等.氯化锂预先给药对异氟醚麻醉诱发老龄大鼠认知功能障碍及海马炎性反应的影响[J].中华麻醉学杂志,2013,33(1):24-27
11胡茂林,宗小芬,梁炜,等.抑郁症模型大鼠海马区磷酸化环磷腺苷反应元件结合蛋白1表达水平及神经元和突触超微结构的研究[J].中华精神科杂志,2015,48(2):92-97
12 Zhou L,Zhang Y,Liu RY,et al.Rescue of impaired long-term facilitation at sensorimotor synapses of Aplysia following siRNA knockdown of CREB1[J].J Neurosci,2015,35(4):1617-1726
13 Jr MG,Sarginson JE,Ryan HS,et al.BDNF and CREB1 genetic variants interact to affect antidepressant treatment outcomes in geriatric depression[J].Pharmacogenet Genomics,2013,23(6):301-313
14彭生,杨建平,张晓庆,等.磷酸二酯酶-4抑制剂Ro20-1724对氯胺酮麻醉后学习记忆及大鼠海马c AMP/PKA-CREB-BDNF信号通路的影响[J].中华行为医学与脑科学杂志,2014,23(6):528-530
15孙东,杨静娴,张囡,等.牛蒡苷元干预CREB/BDNF信号通路保护Aβ25-35损伤神经元的研究[J].中华中医药学刊,2014,19(2):288-290
16 Zhang JC,Yao W,Ren Q,et al.Depression-like phenotype bydeletion ofα7 nicotinic acetylcholine receptor:Role of BDNF-TrkB in nucleus accumbens[J].Sci Rep,2016,6(13):36705-36712
17 Liu ME,Huang CC,Chen MH,et al.Effect of the BDNF Val66Met polymorphism on regional gray matter volumes and cognitive function in the Chinese population[J].Neuromolecular Med,2014,16(1):127-136
18 Hajirezaii B,Barari A,Daloii AA.Consumption of Ginkgo biloba extract and swimming training on NMDA and BDNF plasma[J].Development,2015,141(10):2108-2118
19李金凤,徐桂梅,张辰子,等.银杏叶提取物对MPTP诱导帕金森小鼠保护作用的研究[J].中国比较医学杂志,2016,26(1):46-53
20傅亚,谈宗华,胡承波,等.银杏叶提取物对中枢神经系统保护作用的研究进展[J].中国药房,2013,14(23):2196-2199
21鄢云彪,华骏,兰新新,等.银杏内酯注射液对脑缺血再灌注大鼠运动认知功能的影响[J].中药药理与临床,2018,34(1):138-140
22 Jiang H,Zhao PJ,Su D,et al.Paris saponin I induces apoptosis via increasing the Bax/Bcl-2 ratio and caspase-3 expression in gefitinibresistant non-small cell lung cancer in vitro and in vivo[J].Mol Med Rep,2014,9(6):2265-2271