太白楤木总皂苷对小鼠脑缺血/再灌注的脑保护作用机制
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摘要
目的:探讨太白楤木总皂苷(total saponin of Aralia taibaiensis,SAT)对脑缺血/再灌注小鼠的脑保护作用及其机制。方法:C57BL/6小鼠随机分为假手术组、模型组、阳性药依达拉奉组、太白楤木总皂苷组(15,45和135 mg·kg~(-1))。预防给药3 d,末次给药1 h后,大脑中动脉栓塞法(middle cerebral artery occlusion,MCAO)制备缺血/再灌注模型。再灌注24 h后,通过神经功能评分实验观察太白楤木总皂苷对运动功能的影响;采用2,3,5-氯化三苯基四氮唑(TTC)染色法观察脑损伤梗死体积变化;试剂盒法检测脑组织超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性、丙二醛(MDA)含量; ELISA检测脑组织中肿瘤坏死因子(TNF-α)、白介素1β(IL-1β)、细胞间黏附分子-1(ICAM-1)炎症因子含量的变化; Western blot法测定脑组织胞核NF-κB蛋白和胞质NF-κB,I-κBα及pI-κBα蛋白表达变化。结果:与模型组相比,SAT高剂量组显著降低神经功能评分和脑梗死体积(P <0. 05);中、高剂量组小鼠SOD,GSH-Px活性显著升高(P <0. 05),MDA含量均显著降低(P <0. 05); 3个剂量组的脑组织TNF-α,IL-1β和ICAM-1含量显著降低(P <0. 05)。SAT给药处理后,p-IκBα蛋白表达明显降低,NF-κB核转位减少。结论:SAT能够有效减少缺血/再灌注小鼠脑组织氧化损伤、抑制NF-κB信号通路活化及炎性因子表达而发挥对脑组织的保护作用。
Objective: To evaluate the neuroprotective effects of total saponin of Aralia taibaiensis( SAT)on cerebral ischemia/reperfusion injury( CIRI) and explore the underlying mechanisms. Methods: C57 BL/6 mice were randomly divided into sham operation group,model group,edaravone group,SAT groups( 15,45 and 135 mg·kg~(-1)). The model of CIRI was prepared by middle cerebral artery occlusion( MCAO) 1 h after the last administration of prevention medicine for 3 d. After reperfusion for 24 h,the influence of SAT on movement function was evaluated by using neurologic score test. To observe the volume change of cerebral injury infarction with TTC staining method; to observe the activities of SOD and GSH-Px,the levels of MDA in the brain by using kits method; to observe the content of TNF-α,IL-1β,and ICAM-1 in the brain by using ELISA. In addition,expression of NF-κB protein,I-κBα,and pI-κBα protein of the brain tissues were determined by Western blot analysis. Results: Compared with the model group,the high-dose SAT group significantly reduced neurological score and cerebral infarction volume( P < 0. 05),the activities of SOD and GSH-Px were significantly increased in the medium dose group and high dose group( P < 0. 05),and the content of MDA was significantly reduced( P <0. 05). The contents of TNF-α,IL-1β and ICAM-1 in brain tissue of the three dose groups were significantlyreduced( P < 0. 05). Treatment with SAT,the protein expression of p-IκBα was significantly decreased and the nuclear translocation of NF-κB was significantly inhibited. Conclusion: SAT can play the role of brain tissue protection via effectively reduce oxidative damage of CIRI mice,inhibition the NF-κB signaling pathway and expression of inflammatory factors.
Objective: To evaluate the neuroprotective effects of total saponin of Aralia taibaiensis( SAT)on cerebral ischemia/reperfusion injury( CIRI) and explore the underlying mechanisms. Methods: C57 BL/6 mice were randomly divided into sham operation group,model group,edaravone group,SAT groups( 15,45 and 135 mg·kg~(-1)). The model of CIRI was prepared by middle cerebral artery occlusion( MCAO) 1 h after the last administration of prevention medicine for 3 d. After reperfusion for 24 h,the influence of SAT on movement function was evaluated by using neurologic score test. To observe the volume change of cerebral injury infarction with TTC staining method; to observe the activities of SOD and GSH-Px,the levels of MDA in the brain by using kits method; to observe the content of TNF-α,IL-1β,and ICAM-1 in the brain by using ELISA. In addition,expression of NF-κB protein,I-κBα,and pI-κBα protein of the brain tissues were determined by Western blot analysis. Results: Compared with the model group,the high-dose SAT group significantly reduced neurological score and cerebral infarction volume( P < 0. 05),the activities of SOD and GSH-Px were significantly increased in the medium dose group and high dose group( P < 0. 05),and the content of MDA was significantly reduced( P <0. 05). The contents of TNF-α,IL-1β and ICAM-1 in brain tissue of the three dose groups were significantlyreduced( P < 0. 05). Treatment with SAT,the protein expression of p-IκBα was significantly decreased and the nuclear translocation of NF-κB was significantly inhibited. Conclusion: SAT can play the role of brain tissue protection via effectively reduce oxidative damage of CIRI mice,inhibition the NF-κB signaling pathway and expression of inflammatory factors.
引文
[1]王树林,安芳.中药对脑缺血再灌注损伤保护作用及机制研究进展[J].神经药理学报,2014,4(3):39-48.
[2] PERMPOONPUTTANA K,GOVITRAPONG P. The anti-inflammatory effect of melatonin on methamphetamine-induced proinflammatory mediators in human neuroblastoma dopamine SHSY5Y cell lines[J]. Neurotox Res,2013,23(2):189-199.
[3] WICHA P,TOCHARUS J,JANYOU A,et al. Hexahydrocurcumin protects against cerebral ischemia/reperfusion injury,attenuates inflammation,and improves antioxidant defenses in a rat stroke model[J]. PLoS One,2017,12(12):e0189211.
[4] HARARI OA,LIAO JK. NF-κB and innate immunity in ischemic stroke[J]. Ann N Y Acad Sci,2010,1207(1):32-40.
[5]李成全,周健,徐洲,等.太白楤木的化学成分及药理作用研究进展[J].中南药学,2017,15(10):1401-1409.
[6] XI MM,HAI CX,TANG HF,et al. Antioxidant and antiglycation properties of total saponins extracted from traditional Chinese medicine used to treat diabetes mellitus[J]. Phytother Res,2008,22(2):228-237.
[7]杨志福,汤海峰,贾艳艳,等.太白楤木总皂苷对糖尿病小鼠血糖血脂及抗氧化作用的影响[J].解放军药学学报,2008,24(2):110-113.
[8] WENG Y,YU L,CUI J,et al. Antihyperglycemic,hypolipidemic and antioxidant activities of total saponins extracted from Aralia taibaiensis in experimental type 2 diabetic rats[J]. J Ethnopharmacol,2014,152(3):553-560.
[9]胡文军,熊敏娟.太白楤木根皮总皂苷对小鼠的保肝作用[J].第一军医大学分校学报,2000,23(1):21-23.
[10] LI YN,GUO Y,XI MM,et al. Saponins from Aralia taibaiensis attenuate D-galactose-induced aging in rats by activating FOXO3a and Nrf2 pathways[J]. Oxid Med Cell Longev,2015,2014(3):320513-320525.
[11]范妤,郭东艳,宋强,等.太白木总皂苷对胃癌SGC-7901细胞体外抑制作用研究[J].陕西中医学院学报,2013,36(4):104-106.
[12] DUAN JL,YIN Y,CUI J,et al. Chikusetsu saponinⅣa ameliorates cerebral ischemia reperfusion injury in diabetic mice via adiponectin-mediated AMPK/GSK-3βpathway in vivo and in vitro[J]. Mol Neurobiol,2016,53(1):728-743.
[13] YAN JJ,DUAN JL,WU X,et al. Total saponins from Aralia taibaiensis protect against myocardial ischemia/reperfusion injury through AMPK pathway[J]. Int J Mol Med,2015,36(6):1538-1546.
[14]温富春,徐惠波,丁涛,等.龙牙楤木总皂苷抗心肌缺血作用研究[J].世界科学技术,2005,7(Suppl 1):S5-S8.
[15]孙桂波,徐惠波,温富春,等.龙牙楤木总皂苷对缺氧再给氧心肌细胞损伤的保护作用[J].中国药理学通报,2006,22(9):1092-1094.
[16]陈婵娟.基于心肌mito KATP通道龙牙楤木总皂苷对大鼠缺血再灌注心脏的作用及机制研究[D].北京:北京中医药大学,2014.
[17] PARK S,YAMAGUCHI M,ZHOU C,et al. Neurovascular protection reduces early brain injury after subarachnoid hemorrhage[J]. Stroke,2004,35(10):2412-2417.
[18] CHOI KH,KIM HS,PARK MS,et al. Regulation of Caveolin-1expression determines early brain edema after experimental focal cerebral ischemia[J]. Stroke,2016,47(5):1336-1343.
[19] QU HL,ZHAO M,ZHAO SS,et al. Forced limb-use enhanced neurogenesis and behavioral recovery after strake in the aged rats[J]. Neuroscience,2015,286:316-324.
[20] MIYAUCHI T,WEI EP,POVLISHOCK JT. Evidence for the therapeutic efficacy of either mild hypothermia or oxygen radical scavengers after repetitive mild traumatic brain injury[J]. J Neurotrauma,2014,31(8):773-781.
[21] LI W,TAN C,LIU Y,et al. Resveratrol ameliorates oxidative stress and inhibits aquaporin 4 expression following rat cerebral ischemia-reperfusion injury[J]. Mol Med Rep,2015,12(5):7756-7762.
[22] HE Y,XIAO Y,YANG X,et al. SIRT6 inhibits TNF-α-induced inflammation of vascular adventitial fibroblasts through ROS and Akt signaling pathway[J]. Exp Cell Res,2017,357(1):88-97.
[23] CASTELLANOS M,CASTILLO J,GARCIA MM,et al. Inflammation-mediated damage in progressing lacunar infarctions:a potential therapeutic target[J]. Stroke,2002,33(4):982-987.
[24]董为伟.神经保护的基础与临床[M].第1版.北京:科学出版社,2009:222-238.
[25] LUNDQVIST J,YDE CW,LYKKESFELDT AE. 1α,25-dihydroxyvitamin D3 inhibits cell growth and NFκB signaling in tamoxifen-resistant breast cancer cells[J]. Steroids,2014,85:30-35.
[2] PERMPOONPUTTANA K,GOVITRAPONG P. The anti-inflammatory effect of melatonin on methamphetamine-induced proinflammatory mediators in human neuroblastoma dopamine SHSY5Y cell lines[J]. Neurotox Res,2013,23(2):189-199.
[3] WICHA P,TOCHARUS J,JANYOU A,et al. Hexahydrocurcumin protects against cerebral ischemia/reperfusion injury,attenuates inflammation,and improves antioxidant defenses in a rat stroke model[J]. PLoS One,2017,12(12):e0189211.
[4] HARARI OA,LIAO JK. NF-κB and innate immunity in ischemic stroke[J]. Ann N Y Acad Sci,2010,1207(1):32-40.
[5]李成全,周健,徐洲,等.太白楤木的化学成分及药理作用研究进展[J].中南药学,2017,15(10):1401-1409.
[6] XI MM,HAI CX,TANG HF,et al. Antioxidant and antiglycation properties of total saponins extracted from traditional Chinese medicine used to treat diabetes mellitus[J]. Phytother Res,2008,22(2):228-237.
[7]杨志福,汤海峰,贾艳艳,等.太白楤木总皂苷对糖尿病小鼠血糖血脂及抗氧化作用的影响[J].解放军药学学报,2008,24(2):110-113.
[8] WENG Y,YU L,CUI J,et al. Antihyperglycemic,hypolipidemic and antioxidant activities of total saponins extracted from Aralia taibaiensis in experimental type 2 diabetic rats[J]. J Ethnopharmacol,2014,152(3):553-560.
[9]胡文军,熊敏娟.太白楤木根皮总皂苷对小鼠的保肝作用[J].第一军医大学分校学报,2000,23(1):21-23.
[10] LI YN,GUO Y,XI MM,et al. Saponins from Aralia taibaiensis attenuate D-galactose-induced aging in rats by activating FOXO3a and Nrf2 pathways[J]. Oxid Med Cell Longev,2015,2014(3):320513-320525.
[11]范妤,郭东艳,宋强,等.太白木总皂苷对胃癌SGC-7901细胞体外抑制作用研究[J].陕西中医学院学报,2013,36(4):104-106.
[12] DUAN JL,YIN Y,CUI J,et al. Chikusetsu saponinⅣa ameliorates cerebral ischemia reperfusion injury in diabetic mice via adiponectin-mediated AMPK/GSK-3βpathway in vivo and in vitro[J]. Mol Neurobiol,2016,53(1):728-743.
[13] YAN JJ,DUAN JL,WU X,et al. Total saponins from Aralia taibaiensis protect against myocardial ischemia/reperfusion injury through AMPK pathway[J]. Int J Mol Med,2015,36(6):1538-1546.
[14]温富春,徐惠波,丁涛,等.龙牙楤木总皂苷抗心肌缺血作用研究[J].世界科学技术,2005,7(Suppl 1):S5-S8.
[15]孙桂波,徐惠波,温富春,等.龙牙楤木总皂苷对缺氧再给氧心肌细胞损伤的保护作用[J].中国药理学通报,2006,22(9):1092-1094.
[16]陈婵娟.基于心肌mito KATP通道龙牙楤木总皂苷对大鼠缺血再灌注心脏的作用及机制研究[D].北京:北京中医药大学,2014.
[17] PARK S,YAMAGUCHI M,ZHOU C,et al. Neurovascular protection reduces early brain injury after subarachnoid hemorrhage[J]. Stroke,2004,35(10):2412-2417.
[18] CHOI KH,KIM HS,PARK MS,et al. Regulation of Caveolin-1expression determines early brain edema after experimental focal cerebral ischemia[J]. Stroke,2016,47(5):1336-1343.
[19] QU HL,ZHAO M,ZHAO SS,et al. Forced limb-use enhanced neurogenesis and behavioral recovery after strake in the aged rats[J]. Neuroscience,2015,286:316-324.
[20] MIYAUCHI T,WEI EP,POVLISHOCK JT. Evidence for the therapeutic efficacy of either mild hypothermia or oxygen radical scavengers after repetitive mild traumatic brain injury[J]. J Neurotrauma,2014,31(8):773-781.
[21] LI W,TAN C,LIU Y,et al. Resveratrol ameliorates oxidative stress and inhibits aquaporin 4 expression following rat cerebral ischemia-reperfusion injury[J]. Mol Med Rep,2015,12(5):7756-7762.
[22] HE Y,XIAO Y,YANG X,et al. SIRT6 inhibits TNF-α-induced inflammation of vascular adventitial fibroblasts through ROS and Akt signaling pathway[J]. Exp Cell Res,2017,357(1):88-97.
[23] CASTELLANOS M,CASTILLO J,GARCIA MM,et al. Inflammation-mediated damage in progressing lacunar infarctions:a potential therapeutic target[J]. Stroke,2002,33(4):982-987.
[24]董为伟.神经保护的基础与临床[M].第1版.北京:科学出版社,2009:222-238.
[25] LUNDQVIST J,YDE CW,LYKKESFELDT AE. 1α,25-dihydroxyvitamin D3 inhibits cell growth and NFκB signaling in tamoxifen-resistant breast cancer cells[J]. Steroids,2014,85:30-35.