摘要
目的探讨蛋白酶激活受体2(protease activated receptor 2,PAR 2)对大鼠脊髓损伤后胶质瘢痕形成早期(2周)损伤区神经胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)及波形蛋白(Vimentin)表达的作用。方法建立大鼠重型脊髓损伤模型后立即和第2d于损伤局部给予生理盐水NS(model组)、PAR2特异性抑制剂FSLLRY-NH2(FSLLRY-NH2组)、PAR2 siRNA(PAR2 siRNA组)及PAR2特异性激动剂SLIGRL-NH2(SLI GRL-NH2组);对假手术组(sham operation组)大鼠只打开椎板,不进行其他处理。应用免疫荧光标记技术和蛋白质印迹法检测术后第1d、7d及14dGFAP及Vimentin的表达情况,并进行大鼠后肢运动功能Basso Beattie Bres nahan(BBB)评分。结果伤后第7d及第14d,免疫荧光及蛋白印迹检测皆显示FSLLRY-NH2组及PAR2 siRNA组GFAP及Vimentin蛋白的表达均显著低于model组,而SLIGRL-NH2组该两种蛋白的表达则显著高于model组(P皆<0.05)。伤后第14d,FSLLRY-NH2组及PAR2 siRNA组大鼠BBB评分显著高于model组,而SLIGRL-NH2组则显著低于model组(P皆<0.05)。而在伤后第1d,各干预组GFAP及Vimentin蛋白的表达与model组相比未见明显差异;损伤后第1d及第7d,各干预组大鼠BBB评分与model组相比亦未见明显差异(P皆>0.05)。结论激动PAR2能促进大鼠脊髓损伤后胶质瘢痕形成早期损伤区GFAP与Vimentin的表达,而抑制PAR2则可抑制该两种蛋白的表达,改善胶质瘢痕形成早期的功能预后。
Objective To explore the effects of protease activated receptor 2（PAR2） on glial fibrillary acidic protein （GFAP） and vimentin expression at 2 weeks after spinal cord injury （SCI）. Methods The experimental model of SCI was created by extradural compression of the spinal eord（T11-12 level） using an aneurysm clip with a closing force of 90 g for 1 minute, normal saline （model group）, PAR2 inhibitor FSLLRY-NH2 （FSLLRY-NH2 group）, PAR2 agonist SLIGRL-NH2 （SLIGRL-NH2 group） and PAR2 siRNA （PAR2 siRNA group） were injected into the compression location immediately and at the second day after SCI. The sham operation group rats underwent laminectomy without SCI. The expression levels of GFAP and Vimentin were detected using the immunofluorescenee staining and western blotting at 1, 7 and 14 days after SCI. The Basso Beattie Bresnahan （BBB） score was performed at the same time points mentioned above after SCI. Results At 7 and 14 days after SCI, the expression levels of GFAP and Vimentin expression were significantly higher in model group compared with either FSLLRY-NH2 group or PAR2 siRNA group whereas significantly lower compared with SLI- GRL-NH2 group（P〈0.05 for all comparisons）. At the 14 days, the BBB score in model group was significantly lower com-pared with either FSLLRY-NH2 group or PAR2 siRNA group whereas significantly higher compared with SLIGRL-NH2 group （P〈0.05 for all comparisons）. There was no significant difference in GFAP and Vimentin expression between inter-vention groups and model group at the 1 day. There was no significant difference in BBB score between intervention groups and model group at 1 and 7 days （P〉0.05 for all comparisons）. Conclusions： Activated PAR2 can increase the ex- pression levels of GFAP and Vimentin whereas inhibition of PAR2 can decrease the expression levels of these two proteins and improve the outcomes within 2 weeks after SCI.
出处
《中国神经精神疾病杂志》
CAS
CSCD
北大核心
2013年第7期421-426,共6页
Chinese Journal of Nervous and Mental Diseases
基金
重庆市自然科学基金(编号:cstc2012jjA10055)资助
国家临床重点专科建设项目经费(编号:财社[2011]170号)资助
