健脾补土法组方对脑缺血/再灌注损伤大鼠脑组织t-PA、PAI-1、Col Ⅳ的影响

The Effect of Spleen-strengthening Therapy on t-PA and PAI and Col Ⅳ in Rats with Cerebral Ischemia/Reperfusion Injury

目的观察健脾补土法组方对脑缺血/再灌注损伤大鼠t-PA、PAI-1、ColⅣ的影响,探讨其保护神经细胞可能作用机制。方法将72只雄性SD大鼠随机分为假手术组、模型组、依达拉奉组、健脾补土方小剂量组、健脾补土方中剂量组、健脾补土方大剂量组,每组12只。采用线栓法制备MCAO模型,治疗7 d后处死,并在大鼠处死前进行神经功能缺损评分,应用酶联免疫吸附(ELISA)法测定中组织型纤溶酶原激活物(tPA)及组织型纤溶酶原激活物抑制物-1(PAI-1)的变化,运用免疫组化法测定胶原蛋白Ⅳ(ColⅣ)的含量变化。结果与模型组相比,健脾补土小、中、大剂量组神经功能缺损症状评分减少(P<0.05);健脾补土方小、中、大剂量组t-PA表达明显降低(P<0.05或P<0.01);健脾补土方小、中、大剂量组PAI-1表达明显升高(P<0.05或P<0.01);健脾补土方小、中剂量组Col IV表达明显升高(P<0.05)。结论健脾补土法组方能改善脑缺血/再灌注损伤大鼠神经缺损,可能与其下调t-PA及上调PAI-1的表达,从而减少细胞外基质(ECM)的降解,上调ColⅣ的表达有关。

Objective： To study the effect of Spleen-strengthening therapy on t-PA,PAI-1 and Col IV and in rats with cerebral ischemia/reperfusion injury and investigate its possible mechanism. Methods： 72 male SD rats were randomly divided into sham-operation group,modelling group,edaravone group and low-dosed spleen- strengthening group,middle-dosed spleen-strengthening group,high-dosed spleen-strengthening group, 12 in each group： The models of MCAO were induced by middle cerebra/artery occlusion and recanalization executed after treatment of 7 days,and nerve function defect score of the rats was evaluated before death. ELISA was used to detect the expression levels of t-PA and PAI-1. Immunohistochemical staining was used to detect the expression levels of Col IV. Results： Compared with the modelling group, nerve function defect score was obviously decreased（P〈 0.05） in low-dosed spleen-strengthening group, middle-dosed spleen-strengthening group and high- dosed spleen-strengthening group. Compared with the modelling group,the expression of t-PA was obviously decreased （P 〈 0.05 or P 〈 0.01 ） and PAl was obviously increased （P 〈 0.05 or P 〈 0.01 ） in low-dosed spleen- strengthening group, middle-dosed spleen-strengthening group and high-dosed spleen-strengthening group. Compared with the modelling group ,the expression of Col IV was obviously increased（P〈 0.05 ） in low-dosed spleenstrengthening group and middle-dosed spleen-strengthening group. Conclusion： Spleen-strengthening therapy decreases neurological deficit in rats with cerebral ischemia/reperfusion injury. The mechanism may be the inhibition of the expression of t-PA and promotion of the expression of PAI,which promotes the expression of Col Ⅳ and reduces the degradation of ECM.