目的评估抑制泛素羧基末端水解酶L1(UCHL1)对小鼠脑缺血/再灌注损伤的影响。方法将雄性BALB/c小鼠随机分为假手术(sham)组、缺血/再灌注(I/R)组、UCHL1小干扰RNA(siRNA)组和scramble si RNA(control)组,每组10只小鼠。采用小鼠大脑中动...目的评估抑制泛素羧基末端水解酶L1(UCHL1)对小鼠脑缺血/再灌注损伤的影响。方法将雄性BALB/c小鼠随机分为假手术(sham)组、缺血/再灌注(I/R)组、UCHL1小干扰RNA(siRNA)组和scramble si RNA(control)组,每组10只小鼠。采用小鼠大脑中动脉闭塞(MCAO)60 min后再灌注24 h建立I/R模型。其中si RNA组和control组在MCAO前24 h将10μl UCHL1 si RNA或scramble si RNA通过侧脑室注入脑内。采用RTPCR和Western blotting方法检测各组小鼠脑组织中UCHL1表达;采用2,3,5-氯化三苯基四氮唑(TTC)染色法评估各组小鼠脑梗死体积和水肿率;采用神经行为学评分法评估各组小鼠神经症状评分。结果与sham组相比,I/R组缺血半影区UCHL1 m RNA和蛋白水平显著增高(P<0.05);而si RNA组UCHL1蛋白和mRNA表达明显降低(P<0.05);与此同时,I/R组小鼠脑梗死体积、水肿率及神经行为学损伤明显增加,而si RNA组小鼠脑梗死体积和水肿率及神经行为学损伤进一步加重(P<0.05)。结论抑制UCHL1可加重小鼠脑缺血/再灌注损伤。提示,MCAO后诱导UCHL1表达对小鼠脑缺血/再灌注损伤具有保护作用。展开更多
To ascertain whether a coding mutation (Ile93Met) in ubiquitin carbo xy terminal hydrolase (UCH L1) gene plays a role in idiopathic Parkinson's di sease (IPD) Methods Polymerase chain reaction restriction fra...To ascertain whether a coding mutation (Ile93Met) in ubiquitin carbo xy terminal hydrolase (UCH L1) gene plays a role in idiopathic Parkinson's di sease (IPD) Methods Polymerase chain reaction restriction fragment length polymorphism assay (PCR RFLP) was used to distinguish the wild type (two DNA fragments of 34 and 126 bp ) from the variant allele (three fragments of 34, 60 and 66 bp) because the m utation created a new site for restriction endonuclease BsmF1 DNA was isolated from various blood samples using a phenolchloroform extraction Results Ile93Met substitution was found neither in PD patients nor in controls Conclusions Our study suggested that Ile93Met of UCH L1 gene did not influence risk of IPD展开更多
文摘目的评估抑制泛素羧基末端水解酶L1(UCHL1)对小鼠脑缺血/再灌注损伤的影响。方法将雄性BALB/c小鼠随机分为假手术(sham)组、缺血/再灌注(I/R)组、UCHL1小干扰RNA(siRNA)组和scramble si RNA(control)组,每组10只小鼠。采用小鼠大脑中动脉闭塞(MCAO)60 min后再灌注24 h建立I/R模型。其中si RNA组和control组在MCAO前24 h将10μl UCHL1 si RNA或scramble si RNA通过侧脑室注入脑内。采用RTPCR和Western blotting方法检测各组小鼠脑组织中UCHL1表达;采用2,3,5-氯化三苯基四氮唑(TTC)染色法评估各组小鼠脑梗死体积和水肿率;采用神经行为学评分法评估各组小鼠神经症状评分。结果与sham组相比,I/R组缺血半影区UCHL1 m RNA和蛋白水平显著增高(P<0.05);而si RNA组UCHL1蛋白和mRNA表达明显降低(P<0.05);与此同时,I/R组小鼠脑梗死体积、水肿率及神经行为学损伤明显增加,而si RNA组小鼠脑梗死体积和水肿率及神经行为学损伤进一步加重(P<0.05)。结论抑制UCHL1可加重小鼠脑缺血/再灌注损伤。提示,MCAO后诱导UCHL1表达对小鼠脑缺血/再灌注损伤具有保护作用。
文摘To ascertain whether a coding mutation (Ile93Met) in ubiquitin carbo xy terminal hydrolase (UCH L1) gene plays a role in idiopathic Parkinson's di sease (IPD) Methods Polymerase chain reaction restriction fragment length polymorphism assay (PCR RFLP) was used to distinguish the wild type (two DNA fragments of 34 and 126 bp ) from the variant allele (three fragments of 34, 60 and 66 bp) because the m utation created a new site for restriction endonuclease BsmF1 DNA was isolated from various blood samples using a phenolchloroform extraction Results Ile93Met substitution was found neither in PD patients nor in controls Conclusions Our study suggested that Ile93Met of UCH L1 gene did not influence risk of IPD
