清脑益元汤对大鼠脑缺血再灌注损伤TNF-α、IL-8表达的影响
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摘要
目的:观察清脑益元汤对大鼠脑缺血再灌注损伤肿瘤坏死因子(TNF-α)、白细胞介素(IL-8)表达的影响。方法:将大鼠随机分为空白组,假手术组,脑缺血再灌注组,清脑益元汤组4组。除空白组外,各组均按大鼠脑缺血2 h后,再灌注时间点3、6、12、24、48 h分为5个亚组,每组6只。造模成功后取大鼠缺血侧脑组织,采取免疫组化法检测TNF-α、IL-8的表达,并行HE染色镜下观察大鼠脑组织的病理形态变化。结果:HE染色:(1)空白组、假手术组大鼠脑组织病理形态无明显改变;(2)脑缺血再灌注组大鼠神经元结构明显受损,清脑益元汤组较脑缺血再灌注组在各时间点神经元受损减轻。免疫组化法:(1)大鼠MACO术后,与空白组及假手术组比较,脑缺血再灌注组和清脑益元汤组TNF-α、IL-8阳性细胞的表达在各时间点均升高,差异有统计学意义(P<0.01);(2)脑缺血再灌注组TNF-α、IL-8阳性细胞表达在各时间段均高于清脑益元汤组(P<0.05),两组大鼠在缺血再灌注3 h后,TNF-α、IL-8阳性细胞表达开始增加,6 h显著升高,12 h达高峰,在24 h后呈逐步降低的趋势。结论:清脑益元汤对大鼠脑缺血再灌注后TNF-α、IL-8表达有明显的调控作用,能抑制TNF-α、 IL-8阳性细胞的表达,减轻脑缺血再灌注损伤炎性级联反应,发挥脑保护作用。
Objective: To investigate the effect of QinnaoYiyuan Decoction on the expressions of TNF-α and IL-8 after focal cerebral ischemia reperfusion(I/R) rats. Methods: All SD rats were randomly divided into the normal control group, sham operation group, ischemia-reperfusion group and QinnaoYiyuan Decoction group. Except the normal control group, other groups was further divided into 5 subgroups according to 3 h, 6 h, 12 h, 24 h and 48 h after I/R. The brain was stained by HE and observed with optical microscopes. The changes of TNF-α and IL-8 expressions on the ischemic side of in hippocampus were measured by Immunohisto Chemistry. Results: HE staining:(1)The brain tissue of the normal control group and sham operation group had no significant change in pathological morphology rats.(2)Comparing with ischemia-reperfusion group, the QinnaoYiyuan Decoction group damage of nerve cell was reduced at all time. Immunohistochemistry:(1)Comparing with the normal control group and sham operation group, the ischemia-reperfusion group and QinnaoYiyuan Decoction group, the expressions of TNF-α and IL-8 had increased significantly at all time(P<0.01).(2)As compared with the Qinnao Yiyuan Decoction group, the expressions of TNF-α and IL-8 in the ischemia-reperfusion group had increased significantly at all time(P<0.05). After 3 h of I/R, the expressions of TNF-α and IL-8 in the ischemia-reperfusion group and Qinnao Yiyuan Decoction groups had increased gradually. At 6 h,the expressions were increased significantly and reached the zenith at 12 h and became decreased trend at 24 h. Conclusions: Qinnao Yiyuan Decoction can reduce cerebral ischemia reperfusion injury and play a role of brain protection. It plays a neuron-protective role through reducing the expressions of TNF-α and IL-8.
Objective: To investigate the effect of QinnaoYiyuan Decoction on the expressions of TNF-α and IL-8 after focal cerebral ischemia reperfusion(I/R) rats. Methods: All SD rats were randomly divided into the normal control group, sham operation group, ischemia-reperfusion group and QinnaoYiyuan Decoction group. Except the normal control group, other groups was further divided into 5 subgroups according to 3 h, 6 h, 12 h, 24 h and 48 h after I/R. The brain was stained by HE and observed with optical microscopes. The changes of TNF-α and IL-8 expressions on the ischemic side of in hippocampus were measured by Immunohisto Chemistry. Results: HE staining:(1)The brain tissue of the normal control group and sham operation group had no significant change in pathological morphology rats.(2)Comparing with ischemia-reperfusion group, the QinnaoYiyuan Decoction group damage of nerve cell was reduced at all time. Immunohistochemistry:(1)Comparing with the normal control group and sham operation group, the ischemia-reperfusion group and QinnaoYiyuan Decoction group, the expressions of TNF-α and IL-8 had increased significantly at all time(P<0.01).(2)As compared with the Qinnao Yiyuan Decoction group, the expressions of TNF-α and IL-8 in the ischemia-reperfusion group had increased significantly at all time(P<0.05). After 3 h of I/R, the expressions of TNF-α and IL-8 in the ischemia-reperfusion group and Qinnao Yiyuan Decoction groups had increased gradually. At 6 h,the expressions were increased significantly and reached the zenith at 12 h and became decreased trend at 24 h. Conclusions: Qinnao Yiyuan Decoction can reduce cerebral ischemia reperfusion injury and play a role of brain protection. It plays a neuron-protective role through reducing the expressions of TNF-α and IL-8.
引文
[1] 中华医学会神经病学分会.中国缺血性脑卒中和短暂性脑缺血发作二级预防指南[J].中华神经科杂志,2015,48(4):258-273.
[2] 张玮,李树清.糖皮质激素诱导的蛋白激酶在缺血大鼠海马神经元细胞中的保护作用[J].中国老年学杂志,2016,36(6):1300-1301.
[3] 赵霞霞.清脑益元汤对大鼠脑缺血损伤VEGF、PDGF表达的影响[D].南宁:广西中医药大学,2016.
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[7] 祝美珍.清热化瘀Ⅱ号方对急性脑缺血损伤NF-κBp65蛋白及其关键靶因子表达的影响[D].长沙:湖南中医药大学,2008.
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[9] 李华华,娄淑杰.低氧对大鼠大脑皮层细胞表达IL-1β、IL-6和TNF-α mRNA的影响[J].神经解剖学杂志,2013,29(2):141-144.
[10] 张艳红,陈丽丽,王海英,等.急性脑梗死患者血清肿瘤坏死因子-α、白细胞介素-8和瘦素水平变化[J].中国老年学杂志,2012,32(13):2731-2732.
[11] 范茜茜.电针对缺血再灌注脑损伤大鼠炎症细胞因子IL-4、TNF-α的影响[D].南京:南京中医药大学,2017.
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[13] Khan MM,Motto DG,Lentz SR,et al.ADAMTS13 reducesVWF-mediated acute inflammation following focal cerebralischemia in mice[J].Cerebrovasc Dis,2012,33(5):453-460.
[14] 王瑞,颜江,曹健.TNF-α与脑梗死的关系[J].医学理论与实践,2008,21(1):1-4.
[15] 廖彬.加味补阳还五汤对缺血中风急性期TNF-α、IL-6的影响及疗效评价[D].广州:广州中医药大学,2014.
[16] 祝美珍,朱爱,刘泰,等.清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤IL-6及IL-8表达的影响[J].中华中医药杂志,2017,32(7):3136-3139.
[17] 陈淑增,李国前,王杰华,等.瑞舒伐他汀对脑缺血再灌注损伤大鼠脑组织IL-6和IL-8表达的影响[J].四川大学学报(医学版),2014,45(6):1027-1029.
[18] 刘冲.急性脑梗死患者血清IL-8,IL-17,IL-18水平变化[J].中国实用神经疾病杂志,2013,16(3):28-29.
[19] 千玲玲,贾奎.醒脑静注射液对急性脑梗死患者脑保护作用及白介素-6、白介素-8水平的影响[J].中成药,2013,35(8):1633-1636.
[20] 朱爱,祝美珍.补肾活血法治疗缺血性中风概况[J].中医学报,2014,29(3):406-409.
[21] 祝美珍,王琳,胡跃强,等.清热化瘀Ⅱ号方对急性缺血性中风患者神经功能及中医证候评分的影响[J].中西医结合心脑血管病杂志,2011,9(8):950.
[2] 张玮,李树清.糖皮质激素诱导的蛋白激酶在缺血大鼠海马神经元细胞中的保护作用[J].中国老年学杂志,2016,36(6):1300-1301.
[3] 赵霞霞.清脑益元汤对大鼠脑缺血损伤VEGF、PDGF表达的影响[D].南宁:广西中医药大学,2016.
[4] 张乾,毛善平,李涛,等.构建大脑中动脉阻塞脑缺血模型大鼠的类型分析[J].中国组织工程研究与临床康复,2011(24):4391-4394.
[5] Ei L,Pr W SC.Reversible middle cerebral artery:Occlusion without cranieotomy in rats[J] .Stroke,1989,20:84-91.
[6] 施新猷.现代医学实验动物学[M].北京:人民卫生出版社,2000.
[7] 祝美珍.清热化瘀Ⅱ号方对急性脑缺血损伤NF-κBp65蛋白及其关键靶因子表达的影响[D].长沙:湖南中医药大学,2008.
[8] 陈淑增,李国前,王杰华,等.瑞舒伐他汀对脑缺血再灌注损伤大鼠脑组织IL-6和IL-8表达的影响[J].四川大学学报(医学版),2014,45(6):1027-1029.
[9] 李华华,娄淑杰.低氧对大鼠大脑皮层细胞表达IL-1β、IL-6和TNF-α mRNA的影响[J].神经解剖学杂志,2013,29(2):141-144.
[10] 张艳红,陈丽丽,王海英,等.急性脑梗死患者血清肿瘤坏死因子-α、白细胞介素-8和瘦素水平变化[J].中国老年学杂志,2012,32(13):2731-2732.
[11] 范茜茜.电针对缺血再灌注脑损伤大鼠炎症细胞因子IL-4、TNF-α的影响[D].南京:南京中医药大学,2017.
[12] Gutiérrez-Fernández M,Leciana MA,Rodríguez-Frutos B,et al.CDP-choline at high doses is as effective as i.v.thrombolysis in experimental animal stroke[J].Neurol Res,2012,34(7):649-56.
[13] Khan MM,Motto DG,Lentz SR,et al.ADAMTS13 reducesVWF-mediated acute inflammation following focal cerebralischemia in mice[J].Cerebrovasc Dis,2012,33(5):453-460.
[14] 王瑞,颜江,曹健.TNF-α与脑梗死的关系[J].医学理论与实践,2008,21(1):1-4.
[15] 廖彬.加味补阳还五汤对缺血中风急性期TNF-α、IL-6的影响及疗效评价[D].广州:广州中医药大学,2014.
[16] 祝美珍,朱爱,刘泰,等.清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤IL-6及IL-8表达的影响[J].中华中医药杂志,2017,32(7):3136-3139.
[17] 陈淑增,李国前,王杰华,等.瑞舒伐他汀对脑缺血再灌注损伤大鼠脑组织IL-6和IL-8表达的影响[J].四川大学学报(医学版),2014,45(6):1027-1029.
[18] 刘冲.急性脑梗死患者血清IL-8,IL-17,IL-18水平变化[J].中国实用神经疾病杂志,2013,16(3):28-29.
[19] 千玲玲,贾奎.醒脑静注射液对急性脑梗死患者脑保护作用及白介素-6、白介素-8水平的影响[J].中成药,2013,35(8):1633-1636.
[20] 朱爱,祝美珍.补肾活血法治疗缺血性中风概况[J].中医学报,2014,29(3):406-409.
[21] 祝美珍,王琳,胡跃强,等.清热化瘀Ⅱ号方对急性缺血性中风患者神经功能及中医证候评分的影响[J].中西医结合心脑血管病杂志,2011,9(8):950.