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

作　　者：丁实[1] 赵学荣[2] 李宝群[1] 赵亮[1] 周健[1] 毕红东[1] 庞冲[1] 郭娜娜[1] DING Shi;ZHAO Xuerong;LI Baoqun;ZHAO Liang;ZHOU Jian;BIHongdong;PANG Chong;GUO Nana(Department of Pharmacology,Chengde Medical College,Chengde 067000,China;Department of Immunology,Chengde Medical College,Chengde 067000)

机构地区：[1]承德医学院药理学教研室,承德067000 [2]承德医学院免疫学教研室,承德067000

出　　处：《中国实验动物学报》2019年第5期651-657,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：承德医学院校级课题资助项目(201517)~~

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 h3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。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 of108 SPF grade SD rats were randomly divided into the follow groups: sham group,model group (I/R),berberine low,medium and high dose group (BBR-L, BBR-M,BBR-H),and nimodipine pretreatment group (NMDP group). The rats were scored for neurological function after modeling and the area of cerebral infarction was observed by TTC staining.HE staining was used to observe the histopathological 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 in cerebral 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 at24 h.Compared with the model group,the scores of neurological impairment in the BBR group and NMDP group were significantly reduced at24 h and 48 h ( P <0.05).The percentage of infarct 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/R group,while the levels of caspase-3,Bax mRNA and protein decreased significantly.The Bcl-2/Bax ratio in the I/R group decreased significantly compared with the sham group.The Bcl-2/Bax ratio of each treatment group was increased significantly 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

关 键 词：SD大鼠 BCL-2 BECLIN-1 脑缺血再灌注 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]