血府逐瘀汤调控p38MAPK信号通路改善高血压性脑出血大鼠神经功能研究
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摘要
目的:探讨血府逐瘀汤对高血压性脑出血大鼠神经功能的改善作用及对其p38MAPK信号通路的调控机制。方法:取8只WKY大鼠和48只SHR大鼠,前者记为假手术1组,从后者中随机取8只记为假手术2组,剩余40只均在收缩压≥21.3 kPa时以胶原酶+肝素尾状核注射法制备模型,而假手术2组注射生理盐水其余均同法操作。将模型制备成功的大鼠随机分为模型组、阳性药组、血府逐瘀汤低、中、高剂量组。阳性药组给予安宫牛黄丸25 mg/kg灌胃,血府逐瘀汤低、中、高剂量组分别给予血府逐瘀汤5、10、20 mg/kg灌胃,模型组、假手术1和2组分别给予等量生理盐水灌胃,均干预1周。对比干预前后各组大鼠血肿体积、行为学、神经功能变化,观察出血脑组织病理改变,检测各组出血脑组织p38MAPK蛋白表达,计算p-p38MAPK/p38MAPK。结果:与假手术1组比较,假手术2组向左扭转比率、Berderson、Rosenberg评分、脑组织病理改变、p38MAPK、pp38MAPK/p38MAPK均无明显变化。与模型组比较,阳性药组、血府逐瘀汤低、中、高剂量组血肿体积、向左扭转比率、Berderson评分、Rosenberg评分均下降,脑组织病理均减轻,p-p38MAPK/p38MAPK均升高。与阳性药组比较,血府逐瘀汤低剂量组血肿体积、向左扭转比率、Berderson评分、Rosenberg评分均升高,脑组织病理加重,p-p38MAPK/p38MAPK下降;血府逐瘀汤中剂量组和高剂量组血肿体积、向左扭转比率、Berderson评分、Rosenberg评分均下降,脑组织病理均减轻,p-p38MAPK/p38MAPK均升高。结论:血府逐瘀汤中、高剂量能减少高血压性脑出血大鼠的血肿体积,改善行为学和神经功能,减轻病理损伤,推测与上调p38MAPK蛋白的磷酸化水平有关。
Objective:To discuss the improvement of Xuefu Zhuyu tang on the neurological function in rats with hypertensive cerebral hemorrhage and the regulatory mechanism on their p38 MAPK signaling pathway.Methods:A total of eight WKY rats and 48 SHR rats were selected.The eight WKY rats were grouped into the sham operation group 1,and eight SHR rats were randomly selected and grouped into the sham operation group 2.The remaining 40 rats were injected collagenase and heparin into caudate nucleus when systolic pressure was more than or equal to 21.3 kPa to establish models,and the sham operation group 2 was additionally injected saline based on the above operation.Rats successfully modeled were randomly divided into the model group,the positive medicine group,the Xuefu Zhuyu tang groups of low-dose,medium-dose and high-dose.The positive medicine group was given 25 mg/kg of Angong Niuhuang pills by gavage,the Xuefu Zhuyu tang groups of low-dose,medium-dose and high-dose were given 5,10 and 20 mg/kg of Xuefu Zhuyu tang by gavage respectively,and the model group and the sham operation group 1 and 2 were given equivalent saline by gavage respectively.All groups were intervened for one week.The changes of volume of hematoma,behavior and neurological function in rats of each group before and after intervention was compared; the pathological change in hemorrhagic brain tissue was observed; the expression of p38 MAPK protein in hemorrhagic brain tissue in each group was detected; the p-p38 MAPK/p38 MAPK was calculated.Results:Compared with those in the sham operation group 1,there was no significant change being found in comparisons of left deviation ratio,scores of Berderson as well as Rosenberg,pathological change in hemorrhagic brain tissue,p38 MAPK and p-p38 MAPK/p38 MAPK in the sham operation group 2.Compared with those in the model group,the volume of hematoma,left deviation ratio,scores of Berderson as well as Rosenberg in the positive medicine group and the Xuefu Zhuyu tang groups of low-dose,medium-dose and high-dose were decreased,and the pathology in hemorrhagic brain tissue was alleviated,while p-p38 MAPK/p38 MAPK was increased.Compared with those in the positive medicine group,the volume of hematoma,left deviation ratio,scores of Berderson as well as Rosenberg in the Xuefu Zhuyu tang group of low-dose were increased,and the pathology in hemorrhagic brain tissue was aggravated,while p-p38 MAPK/p38 MAPK was decreased; the volume of hematoma,left deviation ratio,scores of Berderson as well as Rosenberg in the Xuefu Zhuyu tang groups of medium-dose and high-dose were decreased,and the pathology in hemorrhagic brain tissue was alleviated,while p-p38 MAPK/p38 MAPK was increased.Conclusion:Xuefu Zhuyu tang of medium-dose and high-dose can reduce the volume of hematoma in rats with hypertensive cerebral hemorrhage,improve their behavior as well as neurological function,and alleviate pathological injury,which may be related to the up-regulation of the phosphorylation level of p38 MAPK protein.
Objective:To discuss the improvement of Xuefu Zhuyu tang on the neurological function in rats with hypertensive cerebral hemorrhage and the regulatory mechanism on their p38 MAPK signaling pathway.Methods:A total of eight WKY rats and 48 SHR rats were selected.The eight WKY rats were grouped into the sham operation group 1,and eight SHR rats were randomly selected and grouped into the sham operation group 2.The remaining 40 rats were injected collagenase and heparin into caudate nucleus when systolic pressure was more than or equal to 21.3 kPa to establish models,and the sham operation group 2 was additionally injected saline based on the above operation.Rats successfully modeled were randomly divided into the model group,the positive medicine group,the Xuefu Zhuyu tang groups of low-dose,medium-dose and high-dose.The positive medicine group was given 25 mg/kg of Angong Niuhuang pills by gavage,the Xuefu Zhuyu tang groups of low-dose,medium-dose and high-dose were given 5,10 and 20 mg/kg of Xuefu Zhuyu tang by gavage respectively,and the model group and the sham operation group 1 and 2 were given equivalent saline by gavage respectively.All groups were intervened for one week.The changes of volume of hematoma,behavior and neurological function in rats of each group before and after intervention was compared; the pathological change in hemorrhagic brain tissue was observed; the expression of p38 MAPK protein in hemorrhagic brain tissue in each group was detected; the p-p38 MAPK/p38 MAPK was calculated.Results:Compared with those in the sham operation group 1,there was no significant change being found in comparisons of left deviation ratio,scores of Berderson as well as Rosenberg,pathological change in hemorrhagic brain tissue,p38 MAPK and p-p38 MAPK/p38 MAPK in the sham operation group 2.Compared with those in the model group,the volume of hematoma,left deviation ratio,scores of Berderson as well as Rosenberg in the positive medicine group and the Xuefu Zhuyu tang groups of low-dose,medium-dose and high-dose were decreased,and the pathology in hemorrhagic brain tissue was alleviated,while p-p38 MAPK/p38 MAPK was increased.Compared with those in the positive medicine group,the volume of hematoma,left deviation ratio,scores of Berderson as well as Rosenberg in the Xuefu Zhuyu tang group of low-dose were increased,and the pathology in hemorrhagic brain tissue was aggravated,while p-p38 MAPK/p38 MAPK was decreased; the volume of hematoma,left deviation ratio,scores of Berderson as well as Rosenberg in the Xuefu Zhuyu tang groups of medium-dose and high-dose were decreased,and the pathology in hemorrhagic brain tissue was alleviated,while p-p38 MAPK/p38 MAPK was increased.Conclusion:Xuefu Zhuyu tang of medium-dose and high-dose can reduce the volume of hematoma in rats with hypertensive cerebral hemorrhage,improve their behavior as well as neurological function,and alleviate pathological injury,which may be related to the up-regulation of the phosphorylation level of p38 MAPK protein.
引文
[1]高忠明,何吉庆,姜海波,等.脑微出血在急性脑血管病患者中的发生率及危险因素调查研究[J].中国预防医学杂志,2017,18(1):44-48.
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[3]李庆盟,李小黎,邵珺,等.血府逐瘀汤及其类生方方证的思考[J].世界中西医结合杂志,2016,11(6):859-862.
[4] WU C H, CHEN C C, LAI C Y, et al.Treatment with TO901317, a synthetic liver X receptor agonist, reduces brain damage andattenuates neuroinflammation in experimental intracerebral hemorrhage[J].J Neuroinflammation,2016,13(1):62.
[5]杨树升.大承气汤及大黄素对大鼠脑出血的治疗作用及对Nrf2/Src/MAPKs通路的影响[D].武汉:湖北中医药大学,2016.
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[8] YANG P,WU J,MIAO L,et al.Platelet-derived growth factor receptor-βregulates vascular smooth muscle cell phenotypic transformation and neuroinflammation after intracerebral hemorrhage in mice[J].Crit Care Med,2016,44(6):390-402.
[9] FUJIMOTO M,SHIBA M,KAWAKITA F,et al.Deficiency of tenascin-C and attenuation of blood-brain barrier disruption following experimental subarachnoid hemorrhage in mice[J].J Neurosurg,2016,124(6):1693-1702.
[10] KAMTCHUM-TATUENE J,ALLALI G,SAJ A,et al. An exploratory cohort study of sensory extinction in acute stroke:prevalence,risk factors,and time course[J].J Neural Transm(Vienna),2017,124(4):483-494.
[11] ZHAI W,CHEN D,SHEN H,et al.A1 adenosine receptorattenuates intracerebral hemorrhage-induced secondary brain injury in rats byactivating the P38-MAPKAP2-Hsp27pathway[J].Mol Brain,2016,9(1):66.
[12] SATOH M,KATO N,TABEI K I,et al.A case of musical anhedonia due to right putaminal hemorrhage:adisconnection syndrome between the auditory cortex and insula[J].Neurocase,2016,22(6):518-525.
[13]周文,董美华,孙西周,等.补阳还五汤配合西药分期治疗急性高血压性脑出血的疗效观察[J].陕西中医,2016,37(8):971-973.
[14]于朝旭,孙实安,朱炎杰,等.补阳还五汤加减联合小剂量甘露醇对高血压性脑出血术后脑水肿患者的影响[J].现代中西医结合杂志, 2017, 26(16):1789-1791.
[15]孟祥杰.通窍活络汤加减配合运动疗法早期干预脑出血偏瘫的临床观察[J].中国中医急症, 2016, 25(10):1978-1980.
[16]廖远生,杨瑞玲,吴成翰,等.脑出血各证型患者细胞自噬及Cathepsin D表达差异研究[J].中国中西医结合杂志,2018,38(2):173-176.
[17]冯照群,赵晓平,范小璇,等.脑出血患者中医体质与急性期首发证型的关系研究[J].实用中西医结合临床,2016,16(10):23-25.
[18]杨琴芳,许毅,秦峰.桃核承气汤对大鼠脑出血急性期Bcl-2、BaX蛋白表达的影响[J].南京中医药大学学报,2009,25(4):281-282.
[19]高俊英.自拟桃仁川芎汤治疗42例脑出血后遗症疗效观察和护理[J].中国中医基础医学杂志,2013,19(1):110.
[20] LU H, NING X, TAO X, et al.MEKK1 associated with neuronal apoptosis following intracerebral hemorrhage[J].Neurochem Res,2016,41(12):3308-3321.
[2]姚海强,崔红生,郭刚,等.王琦运用血府逐瘀汤治验[J].中医杂志,2016,57(5):375-378.
[3]李庆盟,李小黎,邵珺,等.血府逐瘀汤及其类生方方证的思考[J].世界中西医结合杂志,2016,11(6):859-862.
[4] WU C H, CHEN C C, LAI C Y, et al.Treatment with TO901317, a synthetic liver X receptor agonist, reduces brain damage andattenuates neuroinflammation in experimental intracerebral hemorrhage[J].J Neuroinflammation,2016,13(1):62.
[5]杨树升.大承气汤及大黄素对大鼠脑出血的治疗作用及对Nrf2/Src/MAPKs通路的影响[D].武汉:湖北中医药大学,2016.
[6] SONG Y Q, ZOU H L, ZHAO Y J, et al.Activation of p38-mitogen-activated protein kinase contributes to ischemia reperfusion in ratbrain[J].Genet Mol Res, 2016, 15(3):215-220.
[7] YUAN B,FU F,HUANG S,et al.C5a/C5aR pathway plays a vital role in brain inflammatory injury via initiating Fgl-2 in intracerebral hemorrhage[J].Mol Neurobiol, 2017, 54(8):6187-6197.
[8] YANG P,WU J,MIAO L,et al.Platelet-derived growth factor receptor-βregulates vascular smooth muscle cell phenotypic transformation and neuroinflammation after intracerebral hemorrhage in mice[J].Crit Care Med,2016,44(6):390-402.
[9] FUJIMOTO M,SHIBA M,KAWAKITA F,et al.Deficiency of tenascin-C and attenuation of blood-brain barrier disruption following experimental subarachnoid hemorrhage in mice[J].J Neurosurg,2016,124(6):1693-1702.
[10] KAMTCHUM-TATUENE J,ALLALI G,SAJ A,et al. An exploratory cohort study of sensory extinction in acute stroke:prevalence,risk factors,and time course[J].J Neural Transm(Vienna),2017,124(4):483-494.
[11] ZHAI W,CHEN D,SHEN H,et al.A1 adenosine receptorattenuates intracerebral hemorrhage-induced secondary brain injury in rats byactivating the P38-MAPKAP2-Hsp27pathway[J].Mol Brain,2016,9(1):66.
[12] SATOH M,KATO N,TABEI K I,et al.A case of musical anhedonia due to right putaminal hemorrhage:adisconnection syndrome between the auditory cortex and insula[J].Neurocase,2016,22(6):518-525.
[13]周文,董美华,孙西周,等.补阳还五汤配合西药分期治疗急性高血压性脑出血的疗效观察[J].陕西中医,2016,37(8):971-973.
[14]于朝旭,孙实安,朱炎杰,等.补阳还五汤加减联合小剂量甘露醇对高血压性脑出血术后脑水肿患者的影响[J].现代中西医结合杂志, 2017, 26(16):1789-1791.
[15]孟祥杰.通窍活络汤加减配合运动疗法早期干预脑出血偏瘫的临床观察[J].中国中医急症, 2016, 25(10):1978-1980.
[16]廖远生,杨瑞玲,吴成翰,等.脑出血各证型患者细胞自噬及Cathepsin D表达差异研究[J].中国中西医结合杂志,2018,38(2):173-176.
[17]冯照群,赵晓平,范小璇,等.脑出血患者中医体质与急性期首发证型的关系研究[J].实用中西医结合临床,2016,16(10):23-25.
[18]杨琴芳,许毅,秦峰.桃核承气汤对大鼠脑出血急性期Bcl-2、BaX蛋白表达的影响[J].南京中医药大学学报,2009,25(4):281-282.
[19]高俊英.自拟桃仁川芎汤治疗42例脑出血后遗症疗效观察和护理[J].中国中医基础医学杂志,2013,19(1):110.
[20] LU H, NING X, TAO X, et al.MEKK1 associated with neuronal apoptosis following intracerebral hemorrhage[J].Neurochem Res,2016,41(12):3308-3321.