缝隙连接反义寡核苷酸对大鼠局灶性脑缺血后缝隙连接蛋白表达的影响  

The effect of antisense oligonucleotide on the expression of connectin protein in Focal Cerebral Ischemia Model Rats

作　　者：刘煜[1] 周辉 林宏[1] 林海 

机构地区：[1]第四军医大学唐都医院神经内科,陕西西安710038 [2]西安市中医院神经内科,陕西西安710001

出　　处：《中风与神经疾病杂志》2013年第9期777-780,共4页Journal of Apoplexy and Nervous Diseases

基　　金：陕西省教育厅专项科研计划项目(项目编号06JK310)

摘　　要：目的观察Cx43与Cx32反义寡核苷酸对大鼠局灶性脑缺血后星形胶质细胞Cx43及神经元Cx32表达的影响。方法设计、合成Cx43、Cx32反义寡核苷酸,Cx43、Cx32正义寡核苷酸。正常SD大鼠侧脑室旁立体定位注射Cx43正义寡核苷酸、Cx43反义寡核苷酸,Cx32正义寡核苷酸、Cx32反义寡核苷酸、无菌生理盐水(NS),24h后行颈外动脉插入线栓脑缺血手术,经心冲灌固定取脑,免疫组化法检测Cx43、Cx32蛋白表达;RT-PCR及Western-blot分析Cx43mRNA、Cx32mRNA表达情况。结果在mRNA和蛋白水平上,模型组星形胶质细胞Cx43、神经元Cx32的量表达明显增加;侧脑室注射ACx43、ACx32大鼠,星形胶质细胞与神经元Cx43、Cx32表达均下降,与模型组比较均有统计学差异(P<0.05);侧脑室注射SCx43、SCx32及NS的大鼠脑缺血后,Cx43与Cx32在mRNA与蛋白水平上表达无明显变化,与模型组比较均无统计学差异(P>0.05)。结论 Cx43、Cx32反义寡核苷酸能够抑制Cx43与Cx32在大鼠脑缺血后星形胶质细胞及神经元中的表达。Objective To observe the effect of Cx43 and Cx32 antisense oligonucleotide on the expression of Cx43 and Cx32 in astrocytes and neurons in the rats with focal cerebral ischemia. Methods ACx43, SCx43 antisense oligonucle- otide and sense oligonucleotide were designed and composed with company. The antisense and sense oligonucleotides ACx43 and SCx43 ,normal saline （NS） were injected respectively into the brain beside lateral ventriculus of SD rats. At the same time,the needle was inserted into rat brain without injection （NSNO） as control. The focal cerebral ischemia of rats were constructed by external carotid artery （ECA） with nylon suture 24 hours later. Then the brains of rats were cut into serial eryo-sections after perfused-fixation via heart. The expression of Cx43 and Cx32 protein were detected by immunohis- tochemistry method. RT-PCR and western blot were used to assess the expression amount of Cx43 and Cx32 in ischemia pe- numbra of different groups. Results The expression of Cx43 and Cx32 were obviously increased in mRNA and protein lev- el with ischemia penumbra group. The expression of Cx43 and Cx32 were obviously decreased in mRNA and protein level after Cx43 and Cx32 antisense oligonucleotides were injected into the brains of rats. But expression of Cx43 and Cx32 were not altered at all in the Cx43 and Cx32 sense oligonucleotides injection, saline injection and shame injection rats. Conclu- sion In mRNA and protein level, Cx43 and ACx32 could down-regulate the expression of astrocytes and neurons in focal cerebral ischemia rats.

关 键 词：反义寡核苷酸 脑缺血 缝隙连接蛋白 星形胶质细胞 

分 类 号：R743[医药卫生—神经病学与精神病学]