参麦对急性脑梗死小鼠脑组织的保护作用及相关机制的探讨
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摘要
目的研究参麦注射液对急性脑梗死小鼠脑组织保护作用并初步探讨其可能的机制。方法本实验以30只成年雄性CD-1小鼠为研究对象,随机分为对照组、模型组和实验组,每组各10只。采用线栓法构建急性脑梗死小鼠模型,实验组给予0.3 mL参麦注射液治疗,对照组和模型组均给予同等剂量生理盐水干预,比较各组小鼠的红细胞功能指标。采用TUNEL法测定脑组织的神经元凋亡率,使用Western blot和q-PCR技术检测小鼠脑皮层calpain-1及Bcl-2的表达情况。结果与对照组比较,实验组RBC-C3bR明显降低,RBC-ICR显著升高(P<0.05)。与模型组比较,实验组RBC-C3bR明显增高,RBC-ICR显著降低(P<0.05)。模型组和实验组小鼠脑组织切片的TUNEL阳性细胞数显著高于对照组(P<0.05);实验组小鼠脑组织切片的TUNEL阳性细胞数明显低于模型组小鼠(P<0.05)。与对照组小鼠比较,模型组和实验组小鼠的calpain-1蛋白和mRNA表达水平显著升高,Bcl-2蛋白和mRNA表达水平显著降低(P<0.05);与模型组小鼠相比,实验组小鼠calpain-1蛋白和mRNA表达水平显著降低,Bcl-2蛋白和mRNA表达水平显著升高(P<0.05)。结论参麦注射液可改善急性脑梗死小鼠红细胞功能和抑制神经元凋亡,其可能的机制是下调calpain-1蛋白的表达,上调抗凋亡蛋白Bcl-2的表达,发挥脑组织神经保护作用。
Objective To study the protective effect of a Chinese medicine,Shenmai injection,on brain tissue in a mouse model of cerebral ischemia and explore its possible mechanism.Methods Thirty healthy adult male CD-1 mice were randomly divided into control,model,and experimental groups with 10 mice in each group.The mouse model of acute cerebral infarction was established by permanent middle cerebral artery occlusion.The experimental group was treated with Shenmai injection.The control and model groups were administered the same dose of physiological saline.The red blood cell functional indexes were compared among groups.The neuonal apoptosis rate of brain tissue was determined by TUNEL assay.Calpain-1 and Bcl-2 expression in the mouse cortex was detected by Western blot and q PCR.Results Compared with the control group,RBC-C3 bR was significantly decreased and RBC-ICR was significantly increased in the experimental group(P< 0.05).Compared with the model group,RBC-C3 bR was significantly increased,and RBC-ICR was decreased significantly in the experimental group(P < 0.05).The number of TUNEL-positive cells in the brain tissue sections of model and experimental groups was significantly higher than that of the control group(P< 0.05).The number of TUNELpositive cells in brain tissue sections of the experimental group was significantly lower than that of the model group(P <0.05).Compared with the control group,the expression levels of calpain-1 protein and mRNA in model and experimental groups were significantly increased,and the expression levels of Bcl-2 protein and mRNA were decreased significantly(P<0.05).Compared with the experimental group,the expression levels of calpain-1 protein and mRNA were significantly decreased,and that of Bcl-2 protein and mRNA were increased significantly(P< 0.05).Conclusions Shenmai injection improves erythrocyte functions and inhibites neuronal apoptosis in this mouse model of cerebral infarction.The possible mechanism of the protective effect of Shenmai injection on brain infarction may be the downregulating of expression of calpain-1 protein and upregulating of expression of anti-apoptotic protein Bcl-2 in the brain tissues.
Objective To study the protective effect of a Chinese medicine,Shenmai injection,on brain tissue in a mouse model of cerebral ischemia and explore its possible mechanism.Methods Thirty healthy adult male CD-1 mice were randomly divided into control,model,and experimental groups with 10 mice in each group.The mouse model of acute cerebral infarction was established by permanent middle cerebral artery occlusion.The experimental group was treated with Shenmai injection.The control and model groups were administered the same dose of physiological saline.The red blood cell functional indexes were compared among groups.The neuonal apoptosis rate of brain tissue was determined by TUNEL assay.Calpain-1 and Bcl-2 expression in the mouse cortex was detected by Western blot and q PCR.Results Compared with the control group,RBC-C3 bR was significantly decreased and RBC-ICR was significantly increased in the experimental group(P< 0.05).Compared with the model group,RBC-C3 bR was significantly increased,and RBC-ICR was decreased significantly in the experimental group(P < 0.05).The number of TUNEL-positive cells in the brain tissue sections of model and experimental groups was significantly higher than that of the control group(P< 0.05).The number of TUNELpositive cells in brain tissue sections of the experimental group was significantly lower than that of the model group(P <0.05).Compared with the control group,the expression levels of calpain-1 protein and mRNA in model and experimental groups were significantly increased,and the expression levels of Bcl-2 protein and mRNA were decreased significantly(P<0.05).Compared with the experimental group,the expression levels of calpain-1 protein and mRNA were significantly decreased,and that of Bcl-2 protein and mRNA were increased significantly(P< 0.05).Conclusions Shenmai injection improves erythrocyte functions and inhibites neuronal apoptosis in this mouse model of cerebral infarction.The possible mechanism of the protective effect of Shenmai injection on brain infarction may be the downregulating of expression of calpain-1 protein and upregulating of expression of anti-apoptotic protein Bcl-2 in the brain tissues.
引文
[1]章耀华,马志伟,王海茹.丁苯酞联合丹参酮注射液治疗急性脑梗死临床疗效观察[J].实用老年医学,2017,31(11):1082-1083.
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[14]耿武军,陈成宇,莫云长,等.Wnt/β-catenin信号通路介导参麦注射液对脑缺血再灌注损伤大鼠的脑保护作用[J].中国动脉硬化杂志,2018,26(02):109-114.
[15]臧红敏.α-硫辛酸对电点燃致痫大鼠海马的保护作用及其机制研究[D].石家庄:河北医科大学,2014.
[16]何军娜.瑞替加滨对实验性脑梗死小鼠的脑保护作用:下调calpain-1、Bax,上调Bcl-2的表达[D].石家庄:河北医科大学,2016.
[2]陈红英,郭俐宏,王文杰,等.参麦注射液联合长春西汀治疗脑梗死的临床研究[J].中国医药导报,2015,12(18):126-129.
[3]刘敏,张勤帅,陈学君,等.参麦注射液治疗急性脑梗死有效性和安全性的Meta分析[J].辽宁中医杂志,2017,44(02):228-232.
[4]Tanji N,Kikugawa T,Yokoyama M.Immunohistochemical study of cyclooxygenases in prostatic adenocarcinoma;relationship to apoptosis and Bcl-2 protein expression[J].Anticancer Res,2016,20(4):2313-9.
[5]Liu ZF,Zheng D,Fan GC,et al.Heat stress prevents lipopolysaccharide-induced apoptosis in pulmonary microvascular endothelial cells by blocking calpain/p38 MAPK signalling[J].Apoptosis,2016,21(8):896-904.
[6]Nada SE,Tulsulkar J,Shah ZA.Heme oxygenase 1-mediated neurogenesis is enhanced by Ginkgo biloba(EGb 761)after permanent ischemic stroke in mice[J].Mol Neurobiol,2014,49(2):945-956.
[7]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[8]王渝蓉,余晓霞.化痰祛瘀汤对脑梗死患者的治疗效果及血液流变学、血流动力学的影响[J].环球中医药,2016,9(7):856-858.
[9]李琳,赵丹鹏,刘喜灿,等.参麦注射液对急性缺血性脑卒中患者预后及血清细胞因子水平的影响[J].中药药理与临床,2017,33(2):195-197.
[10]黄一伟,林斌,张明英.参麦注射液联合阿托伐他汀治疗急性心肌梗死临床疗效观察[J].中华中医药学刊,2015,33(10):2558-2560.
[11]Han Z,Li L,Wang L,et al.Alpha-7 nicotinic acetylcholine receptor agonist treatment reduces neuroinflammation,oxidative stress,and brain injury in mice with ischemic stroke and bone fracture[J].J Neurochem,2015,131(4):498-508.
[12]刘育清.丹红注射液联合参麦注射液治疗急性脑梗塞的疗效观察[D].广州:广州中医药大学,2015.
[13]陈中明,刘文兵.参麦注射液辅助治疗对老年急性脑梗死患者红细胞免疫功能、凝血功能及细胞因子的影响[J].中国老年学杂志,2018,38(05):1043-1045.
[14]耿武军,陈成宇,莫云长,等.Wnt/β-catenin信号通路介导参麦注射液对脑缺血再灌注损伤大鼠的脑保护作用[J].中国动脉硬化杂志,2018,26(02):109-114.
[15]臧红敏.α-硫辛酸对电点燃致痫大鼠海马的保护作用及其机制研究[D].石家庄:河北医科大学,2014.
[16]何军娜.瑞替加滨对实验性脑梗死小鼠的脑保护作用:下调calpain-1、Bax,上调Bcl-2的表达[D].石家庄:河北医科大学,2016.