蛇床子素通过抑制线粒体介导的凋亡信号途径减轻脑缺血再灌注损伤
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摘要
目的观察蛇床子素(Osthole)对大鼠脑缺血再灌注损伤的作用及机制。方法将50只SD大鼠随机分成假手术组、模型组和蛇床子素(25、50、100mg/kg)组(n=10)。利用物理法评估大鼠脑梗死范围、脑含水量和神经症状,试剂盒检测活性氧自由基(ROS)和ATP酶活性,流式细胞仪检测线粒体膜电位,免疫蛋白印迹法检测Caspase3、Clevage-Caspase3、Caspase9、Clevage-Caspase9、Bcl-2、Bax、细胞质中凋亡诱导因子(AIF)和cytochrome C的表达水平。结果与假手术组相比,模型组脑梗死范围、脑含水量、神经症状以及ROS、Cleaved-Caspase3、Cleaved-Caspase9、Bax、细胞质中AIF和cytochrome C的表达明显增加,而Caspase3、Caspase9和Bcl-2表达以及线粒体膜电位和ATP酶的活性明显降低。与模型组相比,蛇床子素组脑梗死范围、脑含水量、神经症状以及ROS、Cleaved-Caspase3、CleavedCaspase9、Bax、细胞质中AIF和cytochrome C的表达明显减少,而Caspase3、Caspase9和Bcl-2表达以及线粒体膜电位和ATP酶的活性明显增加。结论蛇床子素预处理可减轻脑缺血再灌注损伤,其机制可能与抑制线粒体介导的凋亡信号途径相关。
Objective To observe the influence of Osthole on cerebral ischemia-reperfusion injury and its acting mechanism.Methods We randomly divided 50 SD rats into sham(Sham)group,ischemia-reperfusion injury(IRI)group and Osthole(25,50,and 100 mg/kg)(Osthole)groups(n=10).The neurological symptoms,extent of cerebral infarction and cerebral water content were evaluated by physical approach.The activities of ROS and ATP were examined by the kit;the mitochondrial membrane potential(MMP)was measured by flow cytometry.The expressions of Bax,Bcl-2,Caspase3,clevage-Caspase3,Caspase9,clevage-Caspase9,cytoplasmic AIF and cytochrome C were measured by Western blotting.Results Compared with those in sham group,the extent of cerebral infarction and cerebral water content,changes of neurological symptoms,activities of ROS,and expressions of clevage-Caspase3,clevage-Caspase9,Bax,cytoplasmic AIF and cytochrome C in IRI group were significantly increased(P<0.05).However,the activities of ATP,expressions of Caspase3,Caspase9,Bcl-2 and MMP,and ATP activity were decreased(P<0.05).Compared with those in IRI group,the extent of cerebral infarction and cerebral water content,changes of neurological symptoms,activities of ROS,and expressions of clevage-Caspase3,clevage-Caspase9,Bax,cytoplasmic AIF and cytochrome C in Osthole group were significantly decreased(P<0.05).But the activities of ATP,expressions of Caspase3,Caspase9,Bcl-2 and MMP,and ATP activities were increased(P <0.05).Conclusion Osthole pretreatment can attenuate cerebral ischemiareperfusion injury,which is associated with suppressing mitochondria-mediated apoptosis.
Objective To observe the influence of Osthole on cerebral ischemia-reperfusion injury and its acting mechanism.Methods We randomly divided 50 SD rats into sham(Sham)group,ischemia-reperfusion injury(IRI)group and Osthole(25,50,and 100 mg/kg)(Osthole)groups(n=10).The neurological symptoms,extent of cerebral infarction and cerebral water content were evaluated by physical approach.The activities of ROS and ATP were examined by the kit;the mitochondrial membrane potential(MMP)was measured by flow cytometry.The expressions of Bax,Bcl-2,Caspase3,clevage-Caspase3,Caspase9,clevage-Caspase9,cytoplasmic AIF and cytochrome C were measured by Western blotting.Results Compared with those in sham group,the extent of cerebral infarction and cerebral water content,changes of neurological symptoms,activities of ROS,and expressions of clevage-Caspase3,clevage-Caspase9,Bax,cytoplasmic AIF and cytochrome C in IRI group were significantly increased(P<0.05).However,the activities of ATP,expressions of Caspase3,Caspase9,Bcl-2 and MMP,and ATP activity were decreased(P<0.05).Compared with those in IRI group,the extent of cerebral infarction and cerebral water content,changes of neurological symptoms,activities of ROS,and expressions of clevage-Caspase3,clevage-Caspase9,Bax,cytoplasmic AIF and cytochrome C in Osthole group were significantly decreased(P<0.05).But the activities of ATP,expressions of Caspase3,Caspase9,Bcl-2 and MMP,and ATP activities were increased(P <0.05).Conclusion Osthole pretreatment can attenuate cerebral ischemiareperfusion injury,which is associated with suppressing mitochondria-mediated apoptosis.
引文
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[13]马淇,刘垒,陈佺.活性氧、线粒体通透性转换与细胞凋亡[J].生物物理学报,2012,7(28):523-536.
[14]刘晓婷,王延让,张明.线粒体介导细胞凋亡的研究进展[J].环境与健康杂志,2013,30(2):182-184.
[15]郑天胜,李翔.线粒体凋亡通路的研究进展[J].2013,9(18):3282-3284.
[2]ZHANG X,YAN H,YUAN Y,et al.Cerebral ischemia-reperfusion-induced autophagy protects against neuronal injury by mitochondrial clearance[J].Autophagy,2013,9(9):1321-1333.
[3]GAO HJ,LIU PF,LI PW,et al.Ligustrazine monomer against cerebral ischemia/reperfusion injury[J].Neural Regen Res,2015,10(5):832-840.
[4]任倩倩,韩冲芳,李婧,等.七氟醚后处理对大鼠脑缺血-再灌注损伤海马血红素氧合酶-1蛋白表达的影响[J].临床麻醉学杂志,2013,29(2):181-184.
[5]SARKHAIL P.Traditional uses,phytochemistry and pharmacological properties of the genus Peucedanum:a review[J].J Ethnopharmacol,2014,156:235-270.
[6]周则卫,沈秀.蛇床子素药理活性的研究概况[J].中国新药杂志,2006,15(20):1727-1730.
[7]LI F,GONG Q,WANG L,et al.Osthole attenuates focal inflammatory reaction following permanent middle cerebral artery occlusion in rats[J].Biol Pharm Bull,2012,35(10):1686-1690.
[8]HE H,WANG S,TIAN J,et al.Protective effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside in the MPTP-induced mouse model of Parkinson's disease:Involvement of reactive oxygen species-mediated JNK,P38 andmitochondrial pathways[J].Eur J Pharmacol,2015,767(5):175-182.
[9]HU Y,DENG H,XU S,et al.MicroRNAs regulate mitochondrial function in cerebral ischemia-reperfusion injury[J].Int J Mol Sci,2015,16(10):24895-24917.
[10]KIM SJ,CHERESH P,JABLONSKI RP,et al.The role of mitochondrial DNA in mediating alveolar epithelial cell apoptosis and pulmonary fibrosis[J].Int J Mol Sci,2015,16(9):21486-214519.
[11]WU N,LI WN,SHU WQ,et al.Blocking the mitochondrial permeability transition pore with cyclosporine-A can restore cardioprotection of ischemic postconditioning in hypercholesterolemic rat heart[J].Eur Rev Med Pharmacol Sci,2015,19(3):446-454.
[12]MORCIANO G,GIORGI C,BONORA M,et al.Molecular identity of the mitochondrial permeability transition pore and its role in ischemia-reperfusion injury[J].J Mol Cell Cardiol,2015,78(2):142-153.
[13]马淇,刘垒,陈佺.活性氧、线粒体通透性转换与细胞凋亡[J].生物物理学报,2012,7(28):523-536.
[14]刘晓婷,王延让,张明.线粒体介导细胞凋亡的研究进展[J].环境与健康杂志,2013,30(2):182-184.
[15]郑天胜,李翔.线粒体凋亡通路的研究进展[J].2013,9(18):3282-3284.