清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤MMP-2及MMP-9表达的影响

Effects of Qingre Huayu Prescription Ⅱ on MMP-2,MMP-9 in rats with cerebral ischemia-reperfusion injury

目的:观察清热化瘀Ⅱ号方对大鼠脑缺血再灌注损伤MMP-2、MMP-9表达的影响。方法:将实验大鼠按随机数字表分为4组,空白组、假手术组、模型组、清热化瘀Ⅱ号方组,模型组和清热化瘀Ⅱ号方组分为I/R 2h后3h、6h、24h、48h、72h 5个亚组,清热化瘀Ⅱ号方组每个时间点又分为低、中、高剂量组。预给药2周后,采用线栓法阻断大脑中脉制备局灶性I/R模型。检测脑组织伊文思蓝(EB)的含量及MMP-2、MMP-9的活性。结果:①脑缺血再灌注3h EB含量增加,至48h达高峰,应用清热化瘀Ⅱ号方后EB含量明显减少。②假手术组大鼠脑组织有少量MMP-2表达,模型组3h时即有较大量MMP-2的活性表达,6h表达有所下降。清热化瘀Ⅱ号方治疗组MMP-2活性表达在同一时间点明显低于模型组,差异有统计学意义(P<0.05)。③假手术组大鼠脑组织见少量MMP-9表达,模型组3h表达开始升高,至48h达高峰。清热化瘀Ⅱ号方治疗组MMP-9表达活性在同一时间点相比明显低于模型组,差异有统计学意义(P<0.05)。结论:清热化瘀Ⅱ号方在脑缺血再灌注损伤过程中,可能通过下调MMP-2、MMP-9的活性而发挥脑保护作用。

Objective： To observe the effects of Qingre Huayu Prescription II on matrix metalloproteinase-2 and matrix metalloproteinase-9 in rats with cerebral ischemia-reperfusion injury. Methods： The rats were randomly divided into 4 groups according to random number table, blank control group, sham operation group, model group, Qingre Huayu group. Model group and Qingre Huayu group were divided into five subgroups according to different reperfusion periods （3h, 6h, 24h, 48h and 72h groups） after 2 hours with cerebral ischemia, and Qingre Huayu group were divided into high dosage group, middle dosage group and low dosage group in each time point. Cerebral ischemia models were established by middle cerebral artery occlusion （MCAO） after preliminary dose for 2 weeks. Results： ①The hemispheric water content of the ischemic side gradually increased with reperfusion periods compared with the sham operation group. The contents of hemispheric water and EB increased at 6h with cerebral ischemia- reperfusion, and reached the peak at 48h. Both of hemispheric water and EB were significantly reduced by Qingre Huayu Prescription II. ②A small amount of expression of MMP-2 could be found in brain tissues in sham operation group. A large amount of expression of MMP-2 could be found in model group at 3h and were declined at 6h. The expression of MMP-2 in Qingre Huayu II group was lower significantly than the model group at each time point, there were significant differences （P〈0.05）. ③Low expression of MMP-9 could be found in brain tissues in sham operation group. The expression of MMP-9 in model group began to rise at 3h, and reached the peak at 48h. Compared with model group, the expression of MMP-9 was lower at each time point, there were significant differences （P〈0.05）. Conclusion： Qingre Huayu Prescription I1 had a protective function for cerebral ischemia-reperfusion injury by relieving cerebral edema, and the possible mechanism is related to inhibited expression of MMP-2 and MMP-9.