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丁实,赵学荣,李宝群,赵亮,周健,毕红东,庞冲,郭娜娜.基于Bcl-2/ Beclin-1 复合体探讨黄连素对脑缺血再灌注损伤大鼠模型的保护作用[J].中国实验动物学报,2019,27(5):651~657.
基于Bcl-2/ Beclin-1 复合体探讨黄连素对脑缺血再灌注损伤大鼠模型的保护作用
Protective effect of berberine on the cerebral ischemia-reperfusion injury in rats based on Bcl-2/ Beclin-1 complex
投稿时间:2019-04-29  
DOI:10. 3969 / j.issn.1005-4847. 2019. 05. 016
中文关键词:  SD 大鼠  Bcl-2  Beclin-1  脑缺血再灌注
英文关键词:SD rats  Bcl-2  Beclin-1  cerebral ischemia-reperfusion injury
基金项目:
作者单位E-mail
丁实 承德医学院药理学教研室,承德 067000 dingshilion@ 163.com 
赵学荣 承德医学院免疫学教研室,承德 067000  
李宝群 承德医学院药理学教研室,承德 067000  
赵亮 承德医学院药理学教研室,承德 067000  
周健 承德医学院药理学教研室,承德 067000  
毕红东 承德医学院药理学教研室,承德 067000  
庞冲 承德医学院药理学教研室,承德 067000  
郭娜娜 承德医学院药理学教研室,承德 067000  
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中文摘要:
      目的 基于B 细胞淋巴瘤/ 白血病-2(Bcl-2) / Beclin-1 复合体研究黄连素对大鼠脑缺血再灌注损伤的神经保护作用?方法 SPF 级SD 大鼠108 只,按照随机数字表法分为假手术组(sham 组)?模型组(I/ R)?黄连素低?中?高剂量组(BBR-L?BBR-M?BBR-H),尼莫地平预处理组(NMDP 组),造模后对大鼠进行神经功能评分;TTC 染色观察大鼠脑梗死面积;HE 染色观察大鼠脑组织病理学变化;RT-PCR 法检测脑组织中Bcl-2?caspase-3?Bax mRNA 的表达;Western blot 检测脑皮质中Bcl-2?caspase-3?Bax?Beclin-1 蛋白表达水平?结果 I/ R 组6?24?48 h 3 个时间点的神经功能评分均显著高于Sham 组,24 h 的神经功能评分最高;与模型组比较,BBR 处理组和NMDP 组24?48 h 的神经功能缺损评分显著降低( P < 0. 05);与Sham 组比较,I/ R 组梗死面积百分比?细胞坏死比例显著升高,与I/ R 组比较,BBR 治疗组和NMDP 组显著减少( P < 0. 05);与Sham 组比较,I/ R组Bcl-2?caspase-3?Bax mRNA 和蛋白水平均显著升高;与I/ R 组比较,BBR 治疗组和NMDP 组Bcl-2 mRNA 和蛋白水平均显著升高,caspase-3?Bax mRNA 和蛋白水平均显著降低;与Sham 组比较,I/ R 组Bcl-2/ Bax 比值显著降低;与I/ R 组比较,各治疗组Bcl-2/ Bax 比值均显著上升,且呈剂量依赖性升高;与Sham 组比较,I/ R 组Beclin-1蛋白水平及Bcl-2/ Beclin-1 比值升高( P < 0. 05);与I/ R 组比较,BBR 组和NMDP 组Beclin-1 蛋白水平显著升高( P < 0. 05),Bcl-2/ Beclin-1 比值均降低,且BBR-M 组?BBR-H 组?NMDP 组Bcl-2/ Beclin-1 无显著差异?结论 黄连素对大鼠脑缺血再灌注损伤具有神经保护作用,其可通过调节自噬蛋白与凋亡蛋白的水平,使自噬达到合适水平,发挥神经保护作用?
英文摘要:
      Objective To study the neuroprotective effect of berberine on cerebral ischemia-reperfusion injury in rats based on the B cell lymphoma/ lewkmia-2 (Bcl-2) / Beclin-1 complex. Methods A total of 108 SPF grade SD ratswere randomly divided into the follow groups: sham group, model group (I/ R), berberine low, medium and high dosegroup ( BBR-L, BBR-M, BBR-H), and nimodipine pretreatment group ( NMDP group). The rats were scored forneurological function after modeling and the area of cerebral infarction was observed by TTC staining. HE staining was used to observe the his topathological changes of rat brain, and the expressions of Bcl-2, caspase-3 and Bax in brain tissues were detected by RT-PCR, while western blot was used to detect the expressions of Bcl-2, caspase-3, Bax and Beclin-1 incerebral cortex. Results The neurological function scores in the I/ R group at 6 h, 24 h and 48 h was significantly higher than that in the sham group, with the highest score at 24 h. Compared with the model group, the scores of neurologicalimpairment in the BBR group and NMDP group were significantly reduced at 24 h and 48 h ( P <0. 05). The percentage ofinfarct area and the percentage of cell necrosis in the I/ R group were increased significantly compared with the sham group( P <0. 05), while BBR group and NMDP group decreased significantly compared with the I/ R group ( P <0. 05). Bcl-2,caspase-3, Bax mRNA and protein levels in the I/ R group were significantly increased compared with the sham group. The levels of Bcl-2 mRNA and protein in the BBR group and NMDP group were increased significantly compared with the I/ Rgroup, while the levels of caspase-3, Bax mRNA and protein decreased significantly. The Bcl-2/ Bax ratio in the I/ R groupdecreased significantly compared with the sham group. The Bcl-2/ Bax ratio of each treatment group was increase dsignificantly compared with the I/ R group and was increased in a dose-dependent manner. Beclin-1 protein level and Bcl-2/Beclin-1 ratio were increased in the I/ R group compared with the sham group ( P <0. 05). The levels of Beclin-1 protein inthe BBR group and NMDP group were increased significantly compared with the I/ R group ( P <0. 05), the ratio of Bcl-2/Beclin-1 was decreased, and there was no significant difference in Bcl-2/ Beclin-1 between the BBR-M group, BBR-Hgroup and NMDP group. Conclusions Berberine has a neuroprotective effect on cerebral ischemia-reperfusion injury in rats. Berberine regulates the levels of autophagy protein and apoptotic protein, so that autophagy can reach an appropriate level and play a neuroprotective role.
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