电针对局灶脑缺血再灌注大鼠大脑皮质eNOS mRNA及蛋白、MMP-9蛋白表达的影响

 摘要:目的 探讨电针对局灶脑缺血/再灌注大鼠脑内缺血皮质区eNOS mRNA及蛋白、MMP-9蛋白表达的影响。方法 采用线栓法制备SD大鼠局灶脑缺血/再灌注模型。将SD大鼠随机分为假手术组、模型组、电针组。缺血2h后把模型组和电针组分为再灌注1、3和7d 3个时间点。取双侧“合谷”穴（LI 4）为电针刺激穴位。免疫组化法检测eNOS蛋白的表达；逆转录聚合酶链法检测eNOS mRNA的表达；Western blot法检测eNOS 、MMP-9蛋白的表达。结果 与假手术组比较，模型组、电针组各时间点大鼠缺血大脑皮质区eNOS蛋白及mRNA表达增加显著（P<0.01）；与模型组比较，电针组各时间点eNOS 蛋白及mRNA增加明显（P<0.01）。eNOS蛋白表达于再灌注3d达高峰，假手术组表达为 (0.4291±0.0173)，模型组再灌注3d表达为 (0.7374±0.0252)，电针组表达为 (0.8536±0.0212)，各组比较差异显著（P<0.01）。与假手术组比较，模型组、电针组各时间点大鼠缺血大脑皮质区MMP-9蛋白的表达增加显著（P<0.01）；与模型组比较，电针组MMP-9蛋白的表达于再灌注1d低于模型组（P<0.01），再灌注3、7 d电针组MMP-9蛋白的表达明显高于模型组（P<0.01）。 结论 电针可上调局灶脑缺血/再灌注大鼠缺血皮质区eNOS蛋白及mRNA、MMP-9蛋白的表达。

Effects of electroacupuncture on mRNA,protein expression of eNOS and protein expression of MMP-9 in rats after focal cerebral ischemia/reperfusion

Objective To investigate the effect of electroacupuncture on mRNA and protein expression of eNOS and protein expression of MMP-9 in rats after focal cerebral ischemia/reperfusion.Methods SD rats received filament occlusion of the right middle cerebral artery for 2 hours followed by reperfusion.Rats were randomly divided into control group,model group and EA group.After 2h of the reperfusion,EA was selected at bilateral "Hegu" point(LI 4) in the EA group.Immunohistochemical method was selected to detect the expression of eNOS protein in the cortical ischemic region.RT-PCR method was used to detect the expression of eNOS mRNA.The expression of eNOS and MMP-9 protein was detected by Western blot.Results Compared with the control group,the protein and mRNA expression of eNOS increased significantly in the model and EA groups(P<0.01).Compared with the model group,the mRNA and protein expression of eNOS were increased significantly in EA groups(P<0.01). The protein expression of eNOS arrived its peak in reperfusion 3 d subgroups: control group was 0.4291±0.0173,model group was 0.7374±0.0252,EA group was 0.8536±0.0212,the difference with the three groups had a statistic value(P<0.01).Compared with the control group,the protein expression of MMP-9 was increased significantly in the model and EA groups(P<0.01).The expression of MMP-9 protein deceased significantly in the 1d subgroup in EA(P<0.01),but increased significantly in the 3 d,7 d subgroups(P<0.01).ConclusionsEA treatment up-regulates the protein and mRNA expression of eNOS and the protein expression of MMP-9 in rats after focal cerebral ischemia/reperfusion.