电针对缺血再灌注脑损伤大鼠Bcl-2、Bax基因的影响

发布时间：2018-05-20 15:18

  本文选题：电针 + 脑缺血再灌注　； 参考：《南京中医药大学》2016年硕士论文

【摘要】：目的：通过观察电针对脑缺血再灌注大鼠模型细胞凋亡相关基因Bcl-2、Bax蛋白表达的影响,探讨电针改善脑缺血后细胞凋亡的调节作用,以期为针刺治疗缺血性脑中风的效应机制的阐释提供科学依据。方法：选用SD大鼠按照随机数字表抽取24只为假手术组,余下动物采用改良Longa线栓法建立脑缺血再灌注模型。将造模成功的大鼠进行随机分组,分别为模型组、电针组,每组有效例数为24只。假手术组除了不穿入线栓外,其他处理与模型组相同；模型组制备模型缺血2h后拔出线栓,不进行任何治疗处理；电针组用一次性针灸针选取“大椎”、“百会”穴进行刺激,“百会”穴平刺,“大椎”穴约30。斜刺,深度约1cm,接韩氏HANS-200型电针仪,“百会”穴接负极,“大椎”穴接正极,选择疏密波,频率为2/15Hz,刺激强度以针柄轻微颤动,大鼠不发出叫声为度,连续治疗30min。脑缺血再灌注24h后,参照Julio氏18分制评分法进行神经功能行为学评分；采用干湿重称量法计算脑含水量,脑组织含水量(%)=(湿重一干重)/湿重×100%；采用免疫组化法和Rt-PCR技术分别检测缺损侧大脑皮层Bcl-2. Bax蛋白表达及Bcl-2mRNA、Bax mRNA的表达变化。结果：(1)脑缺血再灌注损伤大鼠神经功能评分明显降低,与假手术组比较差异显著(P0.01)；电针干预治疗能有效提高其评分值,与模型组比较有显著差异(P0.05)。(2)脑缺血再灌注损伤大鼠脑含水量明显高于假手术组(P0.01),与模型组比较电针干预后脑含水量显著降低(P0.01),并接近假手术组(P0.05)。(3)缺损侧大脑皮层Bcl-2、Bax蛋白的表达,模型组均较假手术组明显升高(P0.05和P0.01),其中Bax蛋白的升高幅度大于Bcl-2蛋白,Bcl-2/Bax蛋白的比值减小(P0.01)；电针组与模型组比较,Bcl-2蛋白的表达升高(P0.05), Bax蛋白的表达降低(P0.01), Bcl-2/Bax蛋白的比值升高(P0.01)。(4)缺损侧大脑皮层Bcl-2mRNA、BaxmRNA的表达,模型组均较假手术组明显升高(P0.05),Bcl-2mRNA/Bax mRNA的比值降低(P0.05)；与模型组比较,电针组Bcl-2mRNA的升高不明显(P0.05),但Bax mRNA的降低显著(P0.05),Bcl-2mRNA/BaxmRNA的比值升高(P0.01)。结论：电针对缺血再灌注脑损伤大鼠Bcl-2、Bax基因具有调控作用,提升Bcl-2/Bax的比值,使抗凋亡基因占据优势,降低缺血再灌注区的细胞凋亡,减轻脑水肿,改善神经功能。
[Abstract]:Objective: to observe the effect of electroacupuncture on the expression of apoptosis-related gene Bcl-2nBax in rat model of cerebral ischemia-reperfusion, and to explore the regulatory effect of electroacupuncture on apoptosis after cerebral ischemia. In order to provide scientific basis for the explanation of the effect mechanism of acupuncture on ischemic stroke. Methods: 24 SD rats were selected as sham-operated group according to random number table. The model of cerebral ischemia-reperfusion was established by modified Longa method. The successful rats were randomly divided into model group and electroacupuncture group with 24 effective cases in each group. The sham-operation group had the same treatment as the model group except for not wearing the thread embolus; the model group prepared the model with 2 hours of ischemia and pulled out the thread plug without any treatment; the electroacupuncture group used a one-off acupuncture needle to select "Dazhui" and "Baihui" to stimulate. "Baihui" point flat thorn, "Dazhui" point about 30. Oblique prickles, depth about 1 cm, HANS-200 type electroacupuncture apparatus, "Baihui" point and "Dazhui" point were connected with negative electrode, and "Dazhui" point was connected with positive pole. The frequency of dense wave was 2 / 15 Hz. The stimulation intensity was slightly quivered with needle handle, and the rats did not emit a call for 30 mins. After 24 hours of cerebral ischemia-reperfusion, the neurobehavioral score was evaluated with Julio's 18 score system, and the water content of brain was calculated by using dry and wet weighing method. The contents of water in brain tissue were as follows: (wet weight, dry weight, wet weight 脳 100), immunohistochemical method and Rt-PCR technique were used to detect Bcl-2 in the damaged cerebral cortex. Expression of Bax protein and Bcl-2 mRNA-Bax mRNA. Results the neurological function score of rats with cerebral ischemia-reperfusion injury was significantly lower than that of sham operation group (P 0.01). Compared with the model group, the brain water content of rats with cerebral ischemia-reperfusion injury was significantly higher than that of the sham-operated group, and the brain water content of the model group was significantly lower than that of the model group after electroacupuncture intervention, and close to the expression of Bcl-2Bax protein in the cortical cortex of the sham-operated group. The expression of Bcl-2 / Bax protein in the model group was significantly higher than that in the sham operation group (P 0.05 and P 0.01), in which the increase of Bax protein was greater than that in the Bcl-2 protein Bcl-2 / Bax protein ratio. The expression of Bcl-2 protein in the electroacupuncture group was significantly higher than that in the model group, and the expression of Bax protein was lower than that in the model group (P 0.01, Bcl-2/Bax protein). The expression of Bcl-2mRNA-mRNA-Bax mRNA in the cortical cortex of the defect side was increased with the increase of the ratio of Bcl-2mRNA-mRNA-BaxmRNA. Compared with the model group, the ratio of P0.05mRNA-2mRNA-Bax mRNA in the model group was significantly higher than that in the sham-operated group, but the ratio of P0.05mRNA-2mRNA-Bax mRNA in the electroacupuncture group was not significantly increased than that in the model group, but the decrease of Bax mRNA was significantly higher than that of P0.05mRNA-2mRNA-Bax mRNA. Conclusion: electroacupuncture can regulate the Bcl-2P Bax gene in rats with cerebral ischemia-reperfusion injury, increase the ratio of Bcl-2/Bax, make anti-apoptotic genes dominant, reduce apoptosis in ischemia-reperfusion area, reduce brain edema and improve neural function.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245

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