针刺联合亚低温对CIRI大鼠凋亡相关因子的影响

发布时间：2018-11-11 15:04

【摘要】：目的:探讨针刺联合亚低温方法对脑缺血再灌损伤(cerebral ischemia reperfusion injury,CIRI)大鼠神经功能缺损评分、脑梗死面积比及细胞凋亡相关因子的影响。方法:参照改良的Zea Longa线栓法制作大鼠大脑中动脉闭塞(Middle cerebralartery occlusion,MCAO)局灶脑缺血再灌注模型,60只SD大鼠随机分为空白组、假手术组、模型组、针刺组、亚低温组、针刺联合亚低温组,每组各10只。治疗72h后,进行神经功能缺损评分、使用TTC染色检测梗死面积比、TUNEL染色观察脑细胞凋亡情况,采用免疫组化检测Bcl-2、Bax、Caspase-3的表达。结果:1.成功建立CIRI大鼠模型,与空白组及假手术组比较,各造模组大鼠神经功能缺损评分明显增高,差异有显著统计学意义(P0.01),造模各组间神经功能缺损评分无统计学意义(P0.05);治疗72h后,与模型组比较,各治疗组大鼠神经功能缺损评分降低,差异有统计学意义(P0.05);与针刺联合亚低温组比较,针刺组差异有统计学意义(P0.05),亚低温组无统计学意义(P0.05),但从评分上可发现联合组优于亚低温组。2.与空白组及假手术组比较,各造模组脑梗死面积比增大,差异有统计学意义(P0.05);与模型组比较,各治疗组间脑梗死面积比相对缩小,差异有统计学意义(P0.05);与针刺联合亚低温组比较,空白组、假手术组及针刺组差异有统计学意义(P0.05),模型组、亚低温组有显著统计学意义(P0.01),且针刺联合亚低温组优于针刺组和亚低温组。3.与空白组及假手术组比较,各造模组大鼠Bax、Caspase-3和细胞凋亡的阳性细胞数增多,Bcl-2阳性细胞数减少,差异有统计学意义(P0.05);各治疗组均能不同程度地降低细胞凋亡的阳性细胞数和Bax、Caspase-3水平、升高Bcl-2表达水平,且针刺联合亚低温组优于针刺组和亚低温组,但三组之间差异无统计学意义(P0.05)。结论:1.脑缺血再灌注损伤引起大鼠明显的神经功能缺损症状和体征、脑梗死面积比增大以及缺血区出现凋亡细胞,而针刺督脉大椎、百会、人中穴联合亚低温疗法可以明显改善大鼠神经功能缺损症状和体征,缩小脑梗死面积比及最大限度的抑制脑细胞凋亡。2.针刺联合亚低温治疗可通过改善神经功能缺损、减少脑梗死面积比、降低缺血区阳性细胞数量来实现对脑细胞的保护作用。3.针刺组、亚低温组及针刺联合亚低温组均能改善缺血再灌注损伤引起的一些列不良反应,且针刺联合亚低温组有优于针刺组或亚低温组的趋势。
[Abstract]:Aim: to investigate the effects of acupuncture combined with mild hypothermia on neurological deficit score, cerebral infarct area ratio and apoptosis related factors in (cerebral ischemia reperfusion injury,CIRI rats with cerebral ischemia-reperfusion injury. Methods: according to the modified Zea Longa thread occlusion method, 60 SD rats were randomly divided into blank group, sham-operation group, model group, acupuncture group, mild hypothermia group and focal cerebral ischemia-reperfusion model of middle cerebral artery occlusion (Middle cerebralartery occlusion,MCAO), and 60 SD rats were randomly divided into three groups: control group, sham operation group, model group, acupuncture group and mild hypothermia group. Acupuncture combined with mild hypothermia group, 10 rats in each group. After 72 hours of treatment, the neurological impairment score, the ratio of infarct area by TTC staining, the apoptosis of brain cells by TUNEL staining, and the expression of Bcl-2,Bax,Caspase-3 by immunohistochemistry were measured. The result is 1: 1. The CIRI rat model was successfully established. Compared with the blank group and the sham operation group, the neurological deficit scores in each model group were significantly higher than those in the control group (P0.01), and the difference was statistically significant (P0.01). There was no significant difference in neurological impairment score among the groups (P0.05). After 72 hours of treatment, compared with the model group, the neurological deficit score of each treatment group decreased, the difference was statistically significant (P0.05). Compared with acupuncture combined with mild hypothermia group, acupuncture group had statistical significance (P0.05), mild hypothermia group had no statistical significance (P0.05), but the score of combined group was better than that of mild hypothermia group. 2. 2. Compared with the blank group and the sham-operation group, the ratio of cerebral infarction area increased in each model group, the difference was statistically significant (P0.05); compared with the model group, the diencephalon infarct area ratio of each treatment group was relatively smaller, the difference was statistically significant (P0.05). Compared with acupuncture combined with mild hypothermia group, there were significant differences among blank group, sham operation group and acupuncture group (P0.05). Model group and mild hypothermia group had significant statistical significance (P0.01). And acupuncture combined with mild hypothermia group was superior to acupuncture group and mild hypothermia group. 3. Compared with the blank group and sham-operation group, the number of Bax,Caspase-3 and apoptosis positive cells increased and the number of Bcl-2 positive cells decreased in each model group (P0.05). The number of apoptotic positive cells and the level of Bax,Caspase-3 were decreased and the expression of Bcl-2 was increased in each treatment group, and the acupuncture combined with mild hypothermia group was superior to the acupuncture group and the mild hypothermia group. However, there was no significant difference among the three groups (P0.05). Conclusion: 1. Cerebral ischemia-reperfusion injury induced obvious neurological deficit symptoms and signs, increased cerebral infarct area ratio and apoptotic cells in the ischemic area. The combination of human midpoint and mild hypothermia therapy can obviously improve the symptoms and signs of neural function defect, reduce the area ratio of cerebral infarction and inhibit the apoptosis of brain cells to the maximum extent. 2. Acupuncture combined with mild hypothermia therapy can protect brain cells by improving neural function defect, reducing cerebral infarction area ratio and decreasing the number of positive cells in ischemic area. 3. Acupuncture group, mild hypothermia group and acupuncture combined with mild hypothermia group can improve some adverse reactions caused by ischemia reperfusion injury, and acupuncture combined with mild hypothermia group has a better trend than acupuncture group or mild hypothermia group.
【学位授予单位】：湖南中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245

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