黄芪甲苷联合迷走神经电刺激对脑缺血再灌注模型大鼠血清氧化应激及炎症因子水平影响研究
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摘要
目的:探讨黄芪甲苷联合迷走神经电刺激(VNS)对脑缺血再灌注模型大鼠血清氧化应激及炎症因子水平影响。方法:选取健康雄性SD大鼠100只,随机分为模型组、电刺激组、黄芪甲苷组、联合组及对照组,各20只。模型组、电刺激组、黄芪甲苷组、联合组大鼠采用改良线栓法建立脑缺血再灌注模型,对照组大鼠接受假手术。模型组和对照组均予以2 mL生理盐水,腹腔注射;电刺激组予以右侧颈部迷走神经电刺激,每次5 min,连续刺激60 min;黄芪甲苷组予以20.0 mg/mL黄芪甲苷溶液2 mL,腹腔注射;联合组予以20.0 mg/mL黄芪甲苷溶液2 mL,腹腔注射+右侧颈部迷走神经电刺激,每次5 min,连续刺激60 min。对大鼠神经功能缺损评分、神经细胞凋亡指数(AI)、脑组织NF-κB蛋白表达水平、氧化应激指标水平及炎症因子水平进行检测。结果:治疗后,对照组、模型组、电刺激组、黄芪甲苷组及联合组大鼠神经功能缺损评分分别为0、(3.41±0.37)、(2.31±0.25)、(2.05±0.29)和(1.58±0.21)分,神经细胞AI分别为(3.29±0.41)、(42.37±5.14)、(31.94±3.49)、(27.55±2.95)和(16.83±2.06)%,NF-κB蛋白表达水平分别为(0.11±0.01)、(0.52±0.07)、(0.43±0.06)、(0.36±0.04)和(0.18±0.02),脑组织超氧化物歧化酶(SOD)水平分别为(18.24±2.19)、(6.23±0.83)、(9.26±1.14)、(11.82±1.27)和(14.38±2.02)U/mg,谷胱甘肽过氧化物酶(GSH-Px)水平分别为(19.11±2.31)、(8.46±1.25)、(11.85±1.37)、(13.58±1.42)和(15.64±2.05)U/mg,过氧化氢酶(CAT)水平分别为(5.69±0.74)、(1.98±0.26)、(2.48±0.32)、(2.94±0.41)和(4.76±0.62)U/mg,丙二醛(MDA)水平分别为(3.98±0.56)、(10.24±1.26)、(8.68±1.12)、(7.11±0.94)和(5.04±0.71)nnol/mg,脑组织肿瘤坏死因子-α(TNF-α)水平分别为(154.37±20.18)、(274.28±31.29)、(245.39±29.74)、(215.85±27.56)和(185.48±22.49)pg/mL,白细胞介素-1β(IL-1β)水平分别为(11.18±1.42)、(32.94±4.16)、(28.65±3.27)、(23.19±2.84)和(17.64±2.35)pg/mL,组间差异有统计学意义(P<0.05)。结论:黄芪甲苷联合VNS能有效减轻脑缺血再灌注大鼠神经功能缺损,抑制神经细胞凋亡,其作用可能与下调脑组织NF-κB蛋白表达,抑制氧化应激及炎症反应有关。
Objective: To analyze the effect of astragaloside combined with vagus nerve stimulation(VNS) on oxidative stress and inflammatory response in rats with cerebral ischemia reperfusion. Methods: A hundred healthy male SD rats were selected and randomly divided into model group, electric stimulation group, astragaloside group, combination group and control group, 20 rats in each. Rats model group, electric stimulation group, astragaloside group and combination group were established cerebral ischemia reperfusion models by modified thread method and rats in the control group underwent sham operation. The model group and the control group were given 2 mL saline by intraperitoneal injection. The electrical stimulation group was given electrical stimulation of right vagus nerve, 5 min for each time, continuous stimulation for 60 min. The astragaloside group was given 20.0 mg/mL astragaloside 2 mL by intraperitoneal injection. The combined group was given 2 mL intraperitoneal injection+ electrical stimulation of right vagus nerve, 5 min for each time, continuous stimulation for 60 min. The neurological deficit scores, neuronal apoptosis index(AI), brain tissue NF-κB protein expression levels, oxidative stress index levels and inflammatory factors levels were detected. Results: After treatment, the neurological deficit scores in control group, model group, electric stimulation group, astragaloside group and combination group were respectively 0,(3.41±0.37),(2.31±0.25),(2.05±0.29) and(1.58±0.21) scores. The neuronal AI values were respectively(3.29±0.41)%,(42.37±5.14)%,(31.94±3.49)%,(27.55±2.95)% and(16.83±2.06)%. The brain tissue NF-κB protein expression levels were respectively(0.11±0.01),(0.52±0.07),(0.43±0.06),(0.36±0.04) and(0.18±0.02). The brain tissue superoxide dismutase(SOD) levels were respectively(18.24±2.19)U/mg,(6.23±0.83)U/mg,(9.26±1.14)U/mg,(11.82±1.27)U/mg and(14.38±2.02)U/mg. The glutathione peroxidase(GSH-Px) levels were respectively(19.11±2.31)U/mg,(8.46±1.25)U/mg,(11.85±1.37)U/mg,(13.58±1.42)U/mg and(15.64±2.05)U/mg. The catalase(CAT) levels were respectively(5.69±0.74)U/mg,(1.98±0.26)U/mg,(2.48±0.32)U/mg,(2.94±0.41)U/mg and(4.76±0.62)U/mg. The malondialdehyde(MDA) levels were respectively(3.98±0.56)nnol/mg,(10.24±1.26)nnol/mg,(8.68±1.12)nnol/mg,(7.11±0.94)nnol/mg and(5.04±0.71)nnol/mg. The brain tissue tumor necrosis factor-α(TNF-α) levels were respectively(154.37±20.18)pg/mL,(274.28±31.29)pg/mL,(245.39±29.74)pg/mL,(215.85±27.56)pg/mL and(185.48±22.49)pg/mL. The interleukin-1β(IL-1β) levels were respectively(11.18±1.42)pg/mL,(32.94±4.16)pg/mL,(28.65±3.27)pg/mL,(23.19±2.84)pg/mL and(17.64±2.35)pg/mL. There was significant difference between the groups(P<0.05). Conclusions: Astragaloside combined with VNS can effectively reduce the neurological deficits in rats with cerebral ischemia reperfusion, inhibit the apoptosis of nerve cells. The effects may be related to down-regulating the expression of NF-κB protein in brain tissue and the inhibition of oxidative stress and inflammatory reaction.
Objective: To analyze the effect of astragaloside combined with vagus nerve stimulation(VNS) on oxidative stress and inflammatory response in rats with cerebral ischemia reperfusion. Methods: A hundred healthy male SD rats were selected and randomly divided into model group, electric stimulation group, astragaloside group, combination group and control group, 20 rats in each. Rats model group, electric stimulation group, astragaloside group and combination group were established cerebral ischemia reperfusion models by modified thread method and rats in the control group underwent sham operation. The model group and the control group were given 2 mL saline by intraperitoneal injection. The electrical stimulation group was given electrical stimulation of right vagus nerve, 5 min for each time, continuous stimulation for 60 min. The astragaloside group was given 20.0 mg/mL astragaloside 2 mL by intraperitoneal injection. The combined group was given 2 mL intraperitoneal injection+ electrical stimulation of right vagus nerve, 5 min for each time, continuous stimulation for 60 min. The neurological deficit scores, neuronal apoptosis index(AI), brain tissue NF-κB protein expression levels, oxidative stress index levels and inflammatory factors levels were detected. Results: After treatment, the neurological deficit scores in control group, model group, electric stimulation group, astragaloside group and combination group were respectively 0,(3.41±0.37),(2.31±0.25),(2.05±0.29) and(1.58±0.21) scores. The neuronal AI values were respectively(3.29±0.41)%,(42.37±5.14)%,(31.94±3.49)%,(27.55±2.95)% and(16.83±2.06)%. The brain tissue NF-κB protein expression levels were respectively(0.11±0.01),(0.52±0.07),(0.43±0.06),(0.36±0.04) and(0.18±0.02). The brain tissue superoxide dismutase(SOD) levels were respectively(18.24±2.19)U/mg,(6.23±0.83)U/mg,(9.26±1.14)U/mg,(11.82±1.27)U/mg and(14.38±2.02)U/mg. The glutathione peroxidase(GSH-Px) levels were respectively(19.11±2.31)U/mg,(8.46±1.25)U/mg,(11.85±1.37)U/mg,(13.58±1.42)U/mg and(15.64±2.05)U/mg. The catalase(CAT) levels were respectively(5.69±0.74)U/mg,(1.98±0.26)U/mg,(2.48±0.32)U/mg,(2.94±0.41)U/mg and(4.76±0.62)U/mg. The malondialdehyde(MDA) levels were respectively(3.98±0.56)nnol/mg,(10.24±1.26)nnol/mg,(8.68±1.12)nnol/mg,(7.11±0.94)nnol/mg and(5.04±0.71)nnol/mg. The brain tissue tumor necrosis factor-α(TNF-α) levels were respectively(154.37±20.18)pg/mL,(274.28±31.29)pg/mL,(245.39±29.74)pg/mL,(215.85±27.56)pg/mL and(185.48±22.49)pg/mL. The interleukin-1β(IL-1β) levels were respectively(11.18±1.42)pg/mL,(32.94±4.16)pg/mL,(28.65±3.27)pg/mL,(23.19±2.84)pg/mL and(17.64±2.35)pg/mL. There was significant difference between the groups(P<0.05). Conclusions: Astragaloside combined with VNS can effectively reduce the neurological deficits in rats with cerebral ischemia reperfusion, inhibit the apoptosis of nerve cells. The effects may be related to down-regulating the expression of NF-κB protein in brain tissue and the inhibition of oxidative stress and inflammatory reaction.
引文
[1] 孟文婷,李东翔,佟玲.缺血性脑卒中的治疗研究进展[J].中国新药杂志,2016,25(10):1114-1120.
[2] 顾美娟,王大力.缺血性脑卒中预后风险述评[J].中国煤炭工业医学杂志,2015,18(2):339-343.
[3] 方健斌,蔡玉洁,崔理立,等.microRNA-146a与缺血性脑卒中的研究进展[J].广东医学,2017,38(9):1435-1437.
[4] 何婵,罗华丽,季晖.基于TNF-α的脑缺血再灌注损伤机制及相关药物研究进展[J].中国药房,2016,27(28):4016-4020.
[5] Jesse D,David P,Anand D,et al.Safety,Feasibility,and Efficacy of Vagus Nerve Stimulation Paired With Upper-Limb Rehabilitation After Ischemic Stroke[J].Stroke,2016,47(1):143-150.
[6] 曲友直,李敏,赵燕玲,等.黄芪甲苷对大鼠脑缺血再灌注后血-脑屏障的保护作用及ZO-1蛋白表达的影响[J].卒中与神经疾病,2016,22(6):278-280.
[7] 田莹,余化霖.线栓法大鼠局灶性脑缺血模型制作要点及进展[J].齐齐哈尔医学院学报,2011,32(2):256-258.
[8] Bederson JB,Pitts LH,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.
[9] 骞秀芳.脑缺血再灌注损伤机制的研究概况[J].基层医学论坛,2015,17(22):3113-3115.
[10] 杨帅,李绪东,钟镝,等.脑缺血再灌注损伤机制的研究进展[J].神经疾病与精神卫生,2015,15(6):620-624.
[11] 王萍,张密霞,庄朋伟,等.脑缺血再灌注损伤的炎症反应机制研究进展[J].天津中医药大学学报,2014,33(5):317-320.
[12] 胡龙涛,蔡芳妮,王亚丽.中风病病因病机探析[J].中西医结合心脑血管病杂志,2017,15(7):883-885.
[13] 曹玉冰.黄芪甲苷的药理作用及其机制的研究进展[J].现代药物与临床,2017,32(5):954-960.
[14] 杨龙,韩冬.黄芪甲苷对局灶性脑缺血再灌注损伤大鼠氧化应激的影响[J].中国中医急症,2016,25(5):799-803.
[15] 魏天祺,冯珍.迷走神经电刺激临床应用及机制研究进展[J].中国康复医学杂志,2015,30(2):185-188.
[16] Ay I,Lu J,Ay H,et al.Vagus nerve stimulation reduces infarct size in rat focal cerebral ischemia[J].Brain Research,2011,459(3):147-151.
[17] Hiraki T,Baker W,Greenberg JH.Effect of vagus nerve stimulation during transient focal cerebral ischemia on chronic outcome in rats[J].Journal of Neuroscience Research,2012,90(4):887-894.
[18] 杨帆,王永青,彭余江,等.NF-κB在颅脑损伤后继发氧化应激及细胞凋亡之间的关系研究[J].浙江创伤外科,2014,21(6):899-902.
[19] 方晓艳,郑艳,贾佼佼,等.脑缺血预处理对脑缺血再灌注损伤大鼠抗氧化酶的影响[J].中药药理与临床,2016,32(3):193-196.
[20] 曲友直,段君民,曹鹏,等.黄芪甲苷对脑缺血再灌注后脑组织MPO活性及TNF-α、IL-1β含量的影响[J].中西医结合心脑血管病杂志,2015,13(14):1620-1621.
[21] 何婵,罗华丽,季晖.基于TNF-α的脑缺血再灌注损伤机制及相关药物研究进展[J].中国药房,2016,27(28):4016-4020.
[22] 李俊杰,蒋海燕,邵建林.脑缺血-再灌注损伤大鼠脑组织中TNF-α,IL-6和IL-1β的表达简[J].昆明医科大学学报,2016,37(9):31-35.
[2] 顾美娟,王大力.缺血性脑卒中预后风险述评[J].中国煤炭工业医学杂志,2015,18(2):339-343.
[3] 方健斌,蔡玉洁,崔理立,等.microRNA-146a与缺血性脑卒中的研究进展[J].广东医学,2017,38(9):1435-1437.
[4] 何婵,罗华丽,季晖.基于TNF-α的脑缺血再灌注损伤机制及相关药物研究进展[J].中国药房,2016,27(28):4016-4020.
[5] Jesse D,David P,Anand D,et al.Safety,Feasibility,and Efficacy of Vagus Nerve Stimulation Paired With Upper-Limb Rehabilitation After Ischemic Stroke[J].Stroke,2016,47(1):143-150.
[6] 曲友直,李敏,赵燕玲,等.黄芪甲苷对大鼠脑缺血再灌注后血-脑屏障的保护作用及ZO-1蛋白表达的影响[J].卒中与神经疾病,2016,22(6):278-280.
[7] 田莹,余化霖.线栓法大鼠局灶性脑缺血模型制作要点及进展[J].齐齐哈尔医学院学报,2011,32(2):256-258.
[8] Bederson JB,Pitts LH,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.
[9] 骞秀芳.脑缺血再灌注损伤机制的研究概况[J].基层医学论坛,2015,17(22):3113-3115.
[10] 杨帅,李绪东,钟镝,等.脑缺血再灌注损伤机制的研究进展[J].神经疾病与精神卫生,2015,15(6):620-624.
[11] 王萍,张密霞,庄朋伟,等.脑缺血再灌注损伤的炎症反应机制研究进展[J].天津中医药大学学报,2014,33(5):317-320.
[12] 胡龙涛,蔡芳妮,王亚丽.中风病病因病机探析[J].中西医结合心脑血管病杂志,2017,15(7):883-885.
[13] 曹玉冰.黄芪甲苷的药理作用及其机制的研究进展[J].现代药物与临床,2017,32(5):954-960.
[14] 杨龙,韩冬.黄芪甲苷对局灶性脑缺血再灌注损伤大鼠氧化应激的影响[J].中国中医急症,2016,25(5):799-803.
[15] 魏天祺,冯珍.迷走神经电刺激临床应用及机制研究进展[J].中国康复医学杂志,2015,30(2):185-188.
[16] Ay I,Lu J,Ay H,et al.Vagus nerve stimulation reduces infarct size in rat focal cerebral ischemia[J].Brain Research,2011,459(3):147-151.
[17] Hiraki T,Baker W,Greenberg JH.Effect of vagus nerve stimulation during transient focal cerebral ischemia on chronic outcome in rats[J].Journal of Neuroscience Research,2012,90(4):887-894.
[18] 杨帆,王永青,彭余江,等.NF-κB在颅脑损伤后继发氧化应激及细胞凋亡之间的关系研究[J].浙江创伤外科,2014,21(6):899-902.
[19] 方晓艳,郑艳,贾佼佼,等.脑缺血预处理对脑缺血再灌注损伤大鼠抗氧化酶的影响[J].中药药理与临床,2016,32(3):193-196.
[20] 曲友直,段君民,曹鹏,等.黄芪甲苷对脑缺血再灌注后脑组织MPO活性及TNF-α、IL-1β含量的影响[J].中西医结合心脑血管病杂志,2015,13(14):1620-1621.
[21] 何婵,罗华丽,季晖.基于TNF-α的脑缺血再灌注损伤机制及相关药物研究进展[J].中国药房,2016,27(28):4016-4020.
[22] 李俊杰,蒋海燕,邵建林.脑缺血-再灌注损伤大鼠脑组织中TNF-α,IL-6和IL-1β的表达简[J].昆明医科大学学报,2016,37(9):31-35.