谷红对大鼠脑出血血肿周围组织NGB和IL-6表达的影响
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摘要
目的:脑红蛋白(neuroglobin,NGB)是德国Burmester等[1]于2000年研究发现一种与氧有很高的亲和力、有利于氧通过血脑屏障和提高脑组织氧的利用率、在脑组织缺氧时有保护作用的蛋白;金特佳(谷红注射液)是谷红复方制剂,综合了乙酰谷酰胺和红花两种药物的有效成分。本实验研究金特佳对脑出血大鼠血肿周围脑组织NGB和IL-6表达的影响,探讨金特佳的脑保护机制,为临床治疗出血性脑血管病提供理论依据。
方法:将72只SD大鼠随机分为3组:实验对照组(n=24),脑出血组(简称出血组,n=24),金特佳药物干预组(简称干预组,n=24),各组在模型建立后12h、24h、48h、72h 4个时间点各随机分为4个亚组。参照李春岩等建立的自体动脉取血并脑内注射的方法复制脑出血模型(尾壳核出血),建立脑出血组和药物干预组,实验对照组在尾状核位置注入生理盐水50μl。药物干预组在出血模型建立后半小时腹腔注射药物金特佳(0.8ml/kg)。分别观察三组动物的以下指标:按Bederson法进行神经功能评价;用干湿重法测定脑出血后血肿周围组织脑含水量的变化,用免疫组织化学方法检测脑出血后不同时间血肿周围脑组织NGB和IL-6的表达。
结果:
1.在出血后相同时间点,药物干预组比出血组神经功能缺损评分明显降低,差异具有统计学意义,P<0.05。
2. 12h时三组大鼠脑含水量无差异,P>0.05,24h时大鼠脑含水量开始升高,实验对照组大鼠与脑出血组大鼠脑含水量比较脑含水量减少,差异具有统计学意义,P<0.05,脑出血组与药物干预组大鼠脑含水量比较脑含水量增多,差异具有统计学意义,P<0.05;48时脑含水量达到高峰,实验对照组与出血组比较差异具有统计学意义,P<0.05,出血组与药物干预组比较无显著差异,P>0.05;水肿高峰持续至72h,实验对照组与出血组比较脑含水量减少,差异具有统计学意义,P<0.05,出血组与药物干预组比较差异无统计学意义;实验对照组各时间点比较无统计学意义,出血组与药物干预组各组不同时间点比较差异具有统计学意义,P<0.05。
3.实验对照组损伤神经元数较出血组少、出血组损伤神经元数较药物干预组增多,差异具有统计学意义,P<0.05。
4.脑出血后12h NGB在血肿周围皮质区即有表达,24h达高峰,72h后数量逐渐下降,同组内各时间点比较有显著差异,P<0.01,各时间点脑出血组与实验对照组比较NGB表达明显增多,差异具有统计学意义P<0.01,干预组与脑出血组比较NGB表达明显增多,差异具有统计学意义P<0.01。
5.脑出血后在血肿周围皮质区可见大量IL-6阳性细胞表达,12h时已经出现并逐渐增多,在48h达到高峰后逐渐减少,组内各时间点比较具有显著差异,P<0.01。各时间点,实验对照组与出血组比较IL-6阳性表达均明显降低,差异具有统计学意义,P<0.05,脑出血组与干预组比较IL-6阳性表达明显增加,差异具有统计学意义P<0.05。
结论:
1.脑出血后,脑出血组较实验对照组血肿周围皮质区NGB表达上调。
2.脑出血后预防性使用金特佳可以对脑出血所致的神经元损伤产生明显的保护作用,表现为减轻神经功能缺损症状,减轻脑水肿,使血肿周围皮质区NGB表达增多,IL-6表达减少。
Objective: To study express pattern of NGB and IL-6 in tissue around cerebral hemorrhage area in rats, investigate effect of Kingtag on express of NGB、IL-6.
Method: 72 male rats(weight 200～250g) were randomly divided into 3 groups : Experimental control group (n=24), Cerebral hemorrhage group (n=24), Drug intervention group(injecting Kingtag abdominal cavity) (n=24), Experiment group and control group were further divided into 4 subgroups respectively according to 12h、24h、48h、72hafter model creation (6 per group).Cerebral hemorrhage model was duplicated with method created by Li Chunyan, namely taking blood from animal femoral artery and injecting into brain. Experimental control group in the caudate nucleus position into saline 50μl. Drug intervention group after the model of bleeding in the half-hour special intraperitoneal injection of the drug Kingtag. Animals in each group were anesthetized by injecting 10%Chloral Hydrate(3.0ml/Kg) via abdominal cavity. Brain was quickly taken after the heart filling with 200ml of saline. Coronal section via caudate nucleus was performed, and express of NGB and IL-6 were assayed with immunohistochemical method. Leukocytic infiltration count and water brain content were also studied. Express of NGB and IL-6 were compared with Experimental control group, Cerebral hemorrhage group and Drug intervention group at different time, and were compared between Cerebral hemorrhage group and Drug intervention group at the same time.
Result: (1) After bleeding in the same time, drug intervention group than the Cerebral hemorrhage group neurological deficit score significantly lower .
(2) Brain water content: cerebral edema worsened 24 hours after hemorrhage and reached the peak at 72 hours. However, degree of cerebral edema in Drug intervention group was lower than that in Cerebral hemorrhage group at the same time.
(3) Experimental control group, bleeding group, drugs damage neurons in the intervention group compared with a few differences.
(4) Express of NGB and IL-6 of tissue around hemorrhage in Cerebral hemorrhage group and Drug intervention group reached the peak at 24 hours. Express of IL-6 reached the peak at 48 hours. Express of NGB keep high degree between 24～48hours. Express of NGB in Cerebral hemorrhage group was obviously lower than that in Drug intervention group. Express of IL-6 in Drug intervention group was obviously lower than that in Cerebral hemorrhage group.
Conclusion:(1) Cerebral hemorrhage, cerebral hemorrhage group than the control group of around hematoma cortex NGB upregulation.
(2) Cerebral hemorrhage after the preventive use of the special can be good for the brain hemorrhage caused by damage neurons have a clear protective effect, the performance in order to alleviate symptoms of neurological deficit, reducing cerebral edema, hematoma surrounding the cortex NGB expression increased, IL-6 expression Reduce.
方法:将72只SD大鼠随机分为3组:实验对照组(n=24),脑出血组(简称出血组,n=24),金特佳药物干预组(简称干预组,n=24),各组在模型建立后12h、24h、48h、72h 4个时间点各随机分为4个亚组。参照李春岩等建立的自体动脉取血并脑内注射的方法复制脑出血模型(尾壳核出血),建立脑出血组和药物干预组,实验对照组在尾状核位置注入生理盐水50μl。药物干预组在出血模型建立后半小时腹腔注射药物金特佳(0.8ml/kg)。分别观察三组动物的以下指标:按Bederson法进行神经功能评价;用干湿重法测定脑出血后血肿周围组织脑含水量的变化,用免疫组织化学方法检测脑出血后不同时间血肿周围脑组织NGB和IL-6的表达。
结果:
1.在出血后相同时间点,药物干预组比出血组神经功能缺损评分明显降低,差异具有统计学意义,P<0.05。
2. 12h时三组大鼠脑含水量无差异,P>0.05,24h时大鼠脑含水量开始升高,实验对照组大鼠与脑出血组大鼠脑含水量比较脑含水量减少,差异具有统计学意义,P<0.05,脑出血组与药物干预组大鼠脑含水量比较脑含水量增多,差异具有统计学意义,P<0.05;48时脑含水量达到高峰,实验对照组与出血组比较差异具有统计学意义,P<0.05,出血组与药物干预组比较无显著差异,P>0.05;水肿高峰持续至72h,实验对照组与出血组比较脑含水量减少,差异具有统计学意义,P<0.05,出血组与药物干预组比较差异无统计学意义;实验对照组各时间点比较无统计学意义,出血组与药物干预组各组不同时间点比较差异具有统计学意义,P<0.05。
3.实验对照组损伤神经元数较出血组少、出血组损伤神经元数较药物干预组增多,差异具有统计学意义,P<0.05。
4.脑出血后12h NGB在血肿周围皮质区即有表达,24h达高峰,72h后数量逐渐下降,同组内各时间点比较有显著差异,P<0.01,各时间点脑出血组与实验对照组比较NGB表达明显增多,差异具有统计学意义P<0.01,干预组与脑出血组比较NGB表达明显增多,差异具有统计学意义P<0.01。
5.脑出血后在血肿周围皮质区可见大量IL-6阳性细胞表达,12h时已经出现并逐渐增多,在48h达到高峰后逐渐减少,组内各时间点比较具有显著差异,P<0.01。各时间点,实验对照组与出血组比较IL-6阳性表达均明显降低,差异具有统计学意义,P<0.05,脑出血组与干预组比较IL-6阳性表达明显增加,差异具有统计学意义P<0.05。
结论:
1.脑出血后,脑出血组较实验对照组血肿周围皮质区NGB表达上调。
2.脑出血后预防性使用金特佳可以对脑出血所致的神经元损伤产生明显的保护作用,表现为减轻神经功能缺损症状,减轻脑水肿,使血肿周围皮质区NGB表达增多,IL-6表达减少。
Objective: To study express pattern of NGB and IL-6 in tissue around cerebral hemorrhage area in rats, investigate effect of Kingtag on express of NGB、IL-6.
Method: 72 male rats(weight 200～250g) were randomly divided into 3 groups : Experimental control group (n=24), Cerebral hemorrhage group (n=24), Drug intervention group(injecting Kingtag abdominal cavity) (n=24), Experiment group and control group were further divided into 4 subgroups respectively according to 12h、24h、48h、72hafter model creation (6 per group).Cerebral hemorrhage model was duplicated with method created by Li Chunyan, namely taking blood from animal femoral artery and injecting into brain. Experimental control group in the caudate nucleus position into saline 50μl. Drug intervention group after the model of bleeding in the half-hour special intraperitoneal injection of the drug Kingtag. Animals in each group were anesthetized by injecting 10%Chloral Hydrate(3.0ml/Kg) via abdominal cavity. Brain was quickly taken after the heart filling with 200ml of saline. Coronal section via caudate nucleus was performed, and express of NGB and IL-6 were assayed with immunohistochemical method. Leukocytic infiltration count and water brain content were also studied. Express of NGB and IL-6 were compared with Experimental control group, Cerebral hemorrhage group and Drug intervention group at different time, and were compared between Cerebral hemorrhage group and Drug intervention group at the same time.
Result: (1) After bleeding in the same time, drug intervention group than the Cerebral hemorrhage group neurological deficit score significantly lower .
(2) Brain water content: cerebral edema worsened 24 hours after hemorrhage and reached the peak at 72 hours. However, degree of cerebral edema in Drug intervention group was lower than that in Cerebral hemorrhage group at the same time.
(3) Experimental control group, bleeding group, drugs damage neurons in the intervention group compared with a few differences.
(4) Express of NGB and IL-6 of tissue around hemorrhage in Cerebral hemorrhage group and Drug intervention group reached the peak at 24 hours. Express of IL-6 reached the peak at 48 hours. Express of NGB keep high degree between 24～48hours. Express of NGB in Cerebral hemorrhage group was obviously lower than that in Drug intervention group. Express of IL-6 in Drug intervention group was obviously lower than that in Cerebral hemorrhage group.
Conclusion:(1) Cerebral hemorrhage, cerebral hemorrhage group than the control group of around hematoma cortex NGB upregulation.
(2) Cerebral hemorrhage after the preventive use of the special can be good for the brain hemorrhage caused by damage neurons have a clear protective effect, the performance in order to alleviate symptoms of neurological deficit, reducing cerebral edema, hematoma surrounding the cortex NGB expression increased, IL-6 expression Reduce.
引文
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